No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Arnica montana effects on gene expression in a human macrophage cell line. Evaluation by quantitative Real-Time PCR
Abstract
Background
Arnica montana is a popular traditional remedy widely used in complementary medicine, also for its wound healing properties. Despite its acknowledged action in clinical settings at various doses, the molecular aspects relating to how A. montana promotes wound healing remain to be elucidated. To fill this gap, we evaluated the whole plant extract, in a wide range of dilutions, in THP-1 human cells, differentiated into mature macrophages and into an alternative IL-4-activated phenotype involved in tissue remodelling and healing.
Methods
Real-time quantitative Reverse Transcription Polymerase Chain Reaction (PCR) analysis was used to study the changes in the expression of a customized panel of key genes, mainly cytokines, receptors and transcription factors.
Results
On macrophages differentiated towards the wound healing phenotype, A. montana affected the expression of several genes. In particular CXC chemokine ligand 1 (CXCL1), coding for an chief chemokine, exhibited the most consistent increase of expression, while also CXC chemokine ligand 2 (CXCL2), Interleukin8 (IL8) and bone morphogenetic protein (BMP2) were slightly up-regulated, suggesting a positive influence of A. montana on neutrophil recruitment and on angiogenesis. MMP1, coding for a metalloproteinase capable of cleaving extracellular matrix substrates, was down-regulated. Most results showed non-linearity of the dose-effect relationship.
Conclusions
This exploratory study provides new insights into the cellular and molecular mechanisms of action of A. montana as a promoter of healing, since some of the genes it modifies are key regulators of tissue remodelling, inflammation and chemotaxis.
... Using various models of macrophage challenge with other micro-organisms, our team has previously shown that highly diluted medicines, instead of killing the micro-organisms, facilitate the organization of a more effective pattern of macrophage response and arrest the parasite's life cycle. [8][9][10][13][14][15][16][17][18][19][20][21][22][23] For this reason, a similar model was designed herein to explain the clinical results, and avoiding new in-vivo experiments. ...
... Cellular mechanisms related to the effects of homeopathic treatments have been well described in recent years and are related to the epigenetic control of specific genes, which depends on the nature of the medicine used and its dilution. 17,18,29,[32][33][34][35] Based on our previous experience, macrophages respond to specific homeopathic stimuli by expressing specific cytokines and chemokines depending on the experimental context. 13,15 The choice of Phos to be used in this study was based on the classical homeopathic similia principle, since the main symptoms of encephalito-zoonosis are listed in Phos materia medica and previous clinical observations reinforced the utility of this medicine for this kind of epidemiological problem. ...
... The findings (1) and (2) above are parasite and/or macrophage features, whose mechanisms need to be elucidated in further studies, though they are probably related to the specific expression of certain genes, as already seen in other similar studies using macrophages invitro. 17,18,[48][49][50] From a practical point of view, putative overlapping between the pro-inflammatory effects of succussed water, including the induction of inflammatory cytokines and chemokines (such as MCP-1, MIP-1β and TNFα) and the specific effects of Phos 200cH (acute expression of RANTES and lysosome activation), would also be present in clinical situations. Thus, an important consideration emerges: perhaps homeopathy acts through different and simultaneous mechanisms, with diverse degrees of specificity, which could be in accordance with the systemic effects observed in clinical practice. ...
Encephalitozoon cuniculi (E. cuniculi) is a fungus that behaves as an intracellular parasite infecting different types of cells. In rabbits and immunosuppressed animals of other species, including humans, it parasites neural tissues causing a highly characteristic neurological syndrome, for which repertory analysis indicates Phosphorus. Successful treatment of 7 naturally infected rabbits with Phosphorus confirmed that this medicine probably is the remedy of the ‘epidemic genius’ of microsporidiosis. In the present study, an in vitro model was used to evidence the intracellular relationship between macrophages and E. cuniculi after treatment of co-cultures with different potencies of Phosphorus. RAW 254.7 macrophages were co-cultivated with E. cuniculi and treated with Phosphorus 6cH, 30cH and 200cH. Controls were untreated co-cultures and cultures treated with vehicle (0.06% final alcohol concentration). Phagocytosis and lysosome activity were after after 1 and 24 hours of incubation by means of the calcofluor and acridine orange staining methods, followed by automatic image analysis (Metamorph®). Cytokine production was assessed with the MAGPIX®-Luminex system. The vehicle increased IL-6, MCP-1 and MIP1 production (p
... Using various models of macrophage challenge with other micro-organisms, our team has previously shown that highly diluted medicines, instead of killing the micro-organisms, facilitate the organization of a more effective pattern of macrophage response and arrest the parasite's life cycle. [8][9][10][13][14][15][16][17][18][19][20][21][22][23] For this reason, a similar model was designed herein to explain the clinical results, and avoiding new in-vivo experiments. ...
... Cellular mechanisms related to the effects of homeopathic treatments have been well described in recent years and are related to the epigenetic control of specific genes, which depends on the nature of the medicine used and its dilution. 17,18,29,[32][33][34][35] Based on our previous experience, macrophages respond to specific homeopathic stimuli by expressing specific cytokines and chemokines depending on the experimental context. 13,15 The choice of Phos to be used in this study was based on the classical homeopathic similia principle, since the main symptoms of encephalito-zoonosis are listed in Phos materia medica and previous clinical observations reinforced the utility of this medicine for this kind of epidemiological problem. ...
... The findings (1) and (2) above are parasite and/or macrophage features, whose mechanisms need to be elucidated in further studies, though they are probably related to the specific expression of certain genes, as already seen in other similar studies using macrophages invitro. 17,18,[48][49][50] From a practical point of view, putative overlapping between the pro-inflammatory effects of succussed water, including the induction of inflammatory cytokines and chemokines (such as MCP-1, MIP-1β and TNFα) and the specific effects of Phos 200cH (acute expression of RANTES and lysosome activation), would also be present in clinical situations. Thus, an important consideration emerges: perhaps homeopathy acts through different and simultaneous mechanisms, with diverse degrees of specificity, which could be in accordance with the systemic effects observed in clinical practice. ...
Introduction Encephalitozoon cuniculi (E. cuniculi), a fungus that acts as an intracellular pathogen, causes a marked neurological syndrome in many host species and is a zoonotic concern. Although no well-established treatment for this syndrome is known, previous successful clinical experience using homeopathic phosphorus has been described in which symptom remission with no mortality occurred in 40/42 animals by means of unknown immunological mechanisms. The latter observation was the main motivation for this study.
Objective To verify, in an in-vitro model, if macrophages infected with E. cuniculi can change in function after treatment with different potencies of phosphorus.
Materials and Methods RAW 264.7 macrophages were infected with E. cuniculi in-vitro and treated with various homeopathic potencies of phosphorus. The vehicle was used as a control solution (0.06% succussed ethanol). After 1 and 24 hours, the following parameters were analyzed: parasite internalization (by the Calcofluor staining method), lysosome activity (by the acridine orange method), cytokine/chemokine production (by the MAGPIX system), and cell ultrastructure. Automatic image analysis was used when applicable, and the experiments were performed in triplicate.
Results Treatment with vehicle alone increased interleukin (IL)-6, tumor necrosis factor alpha and monocyte chemotactic protein -1 production (p ≤ 0.05) and reduced the number of internalized parasites (p ≤ 0.001). A progressive and time-dependent increase in RANTES (regulated on activation, normal T-cell expressed and secreted) and lysosome activity (p ≤ 0.002) was observed only after treatment with the highest potency of phosphorus (Phos 200cH), together with decreased apoptosis rate, intense parasite digestion, and the presence of non-internalized spores.
Conclusions Phos 200 cH has a modulatory action on the activity of infected macrophages, especially a specific increase in RANTES, a key element in the prognosis of E. cuniculi-infected and of immunosuppressed patients bearing infections.
... There are few scientific studies evaluating its effects on osteoporosis and wound healing in the presence of osteoporosis. Some experimental evidence in laboratory animals found an anti-inflammatory action of A. montana [8]. ...
... The action of A. montana as a promoter of healing by means of increasing the expression of several genes and some of these ones are key regulators for tissue remodelling, inflammation and chemotaxis [8]. ...
... The ovariectomized group treated with A. montana 6CH had the best behavior in biomechanical test, while presented similar results in the others analysis. Olioso et al. observed the positive effects of homeophatic use of A. montana and verified that it exhibited a remarkable tendency to increase bone morphogenetic protein (BMP2) which is required for the initiation of bone repair [8]. A. montana homeopathic remedy increases the energy metabolism of cells by protecting the mitochondria from oxidative damage caused by lipid peroxidation [13]. ...
p> Objective: National and international data show that in the next years the elder population tends to have an exponential increase becoming to be significantly higher than other ages. Among the frequent diseases in the elderly, osteoporosis is a disease that decreases bone mass and deteriorates bone structure causing fragility and a high risk of fracture. This study aimed to evaluate the effect of phytotherapic and homeopathic Arnica montana on bone repair quality and its possible use in the treatment of bone fracture in patients with osteoporosis. Material and Methods: This experimental study was performed in Wistar adult female rats divided in 4 groups according to the following treatments: ovariectomized treated with homeopathic A. montana 6CH (OVZ 6CH); ovariectomized treated with phytotherapic A. montana (OVZ TM); ovariectomized with placebo (OVZ PL) and rats with sham surgery and placebo (Sham PL). In a period of 45 days after ovariectomy or sham surgery, all animals got a monocortical lesion. They were euthanized after 6, 12, 18 and 28 days and had the tibiae removed to evaluate dimensions and bone repair by radiographic density, biomechanical test and scanning electronic microscopy (SEM). Data were analyzed by ANOVA and TUKEY tests (p < 0.05). Results: Results showed that treatment with Arnica 6CH was better than Arnica TM considering bone resistance and bone repair quality. Conclusion: This study concluded that treatment with homeopathic A. montana was more efficient than with phytotherapic Arnica in bone regeneration in rats with osteoporosis.
Keywords
Arnica montana ; Homeopathy; Osteoporosis; Phytotherapy; Radiographic bone density. </p
... Furthermore, OCN is a bone protein that is produced by osteoblast cells at the final stage of bone mineralization and is used in clinical practice as a particular biomarker of bone development [23]. According to a previous study by Olioso et al. [24] A. montana increases the activity of IL-4 macrophages in wound healing by stimulating BMP2 expression [24]. In our study, Immunocytochemistry was carried out to check for the expression of OCN, a late differentiation marker. ...
... Furthermore, OCN is a bone protein that is produced by osteoblast cells at the final stage of bone mineralization and is used in clinical practice as a particular biomarker of bone development [23]. According to a previous study by Olioso et al. [24] A. montana increases the activity of IL-4 macrophages in wound healing by stimulating BMP2 expression [24]. In our study, Immunocytochemistry was carried out to check for the expression of OCN, a late differentiation marker. ...
Background
Arnica montana and Bellis perennis are two medicinal plants that are thought to accelerate bone repair in homoeopathic literature. Mesenchymal stem cells (MSCs) are multipotent stem cells with the ability to differentiate and regenerate bone or osteogenesis. Hence, we aimed to determine the role of Arnica montana and Bellis perennis on the osteogenic differentiation of the C3H10T1/2 stem cell line.
Methods and Results
The cell proliferation of Arnica montana and Bellis perennis was evaluated by MTT assay. Osteogenic differentiation of C3H10T1/2 was induced by the addition of β-glycerophosphate, ascorbic acid and dexamethasone in the differentiation medium over 3 weeks. Cells were treated with Arnica montana and Bellis perennis individually as well as in combination. The osteogenic differentiation potential of Arnica montana and Bellis perennis to differentiate C3H10T1/2 into osteoblasts was measured by alkaline phosphatase activity, alizarin red staining and the expression of Osteocalcin using immunostaining and qRT-PCR.
Arnica montana and Bellis perennis could enhance C3H10T1/2 cell proliferation at 1600 µg. Further, the compound showed the ability to augment osteogenesis as confirmed by increased expression of alkaline phosphatase and enhanced calcium accumulation as seen by the Alizarin Red staining and quantification. Enhanced osteogenesis was further supported by the increased expression of osteocalcin in the treated cells with individual and combined doses of Arnica montana and Bellis perennis. Therefore, the findings provide additional support for the positive impact of Arnica montana and Bellis perennis on bone formation.
Conclusions
Our findings suggest that homoeopathic compounds Arnica montana and Bellis perennis can augment osteogenesis individually as well as in combination.
Graphical abstract
... 30 In another study, Arn induced increased expression of the chemokine CXCL1, suggesting positive influence on cell recruitment and angiogenesis, in human macrophage cell lines. 31 The rationale for carrying out the current study was based on the anti-inflammatory properties of Arn, which led us to evaluate its potential against BCa. Moreover, as per homeopathic literature, the disease-producing property in a remedy also contributes to its disease-curing ability. ...
... Moreover, as per homeopathic literature, the disease-producing property in a remedy also contributes to its disease-curing ability. 32 Therefore, using this as the foundation of our study, we hypothesized that if Arn could induce angiogenesis and chemotaxis in normal tissues, 30,31 it would potentially also have the ability to treat diseases like cancer with a similar underlying pathology. ...
Background Breast cancer is the most common cancer in women worldwide. Use of homeopathic medicines for the treatment of cancers has increased in the last several years. Arnica montana is an anti-inflammatory homeopathic medicine used in traumatic conditions and because of this property we performed investigations for its potential as a chemotherapeutic agent against breast cancer.
Methods An ethanolic extract of Arnica montana (mother tincture, MT), prepared according to the Homoeopathic Pharmacopoeia of India, was characterized by gas chromatography–mass spectroscopy (GC–MS), followed by computational (in silico) analysis using molecular docking, to identify specific compounds that can bind and modulate the activity of key proteins involved in breast cancer survival and progression. To validate the in silico findings, in a controlled experiment breast cancer cells (MCF7) were treated in vitro with Arnica montana and the cytotoxic effects assessed by flowcytometry, fluorescence microscopy, scratch assay, clonogenic potential and gene expression analysis.
Results Phytochemical characterization of ethanolic extract of Arn MT by GC–MS allowed identification of several compounds. Caryophyllene oxide and 7-hydroxycadalene were selected for molecular docking studies, based on their potential drug-like properties. These compounds displayed selective binding affinity to some of the recognized target proteins of breast cancer, which included estrogen receptor alpha (ERα), progesterone receptor (PR), epidermal growth factor receptor (EGFR), mTOR (mechanistic target of rapamycin) and E-cadherin. In vitro studies revealed induction of apoptosis in MCF7 cells following treatment with Arn MT. Furthermore, treatment with Arn MT revealed its ability to inhibit migration and colony forming abilities of the cancer cells.
Conclusion Considering the apoptotic and anti-migratory effects of Arnica montana in breast cancer cells in vitro, there is a need for this medicine to be further validated in an in vivo model.
... 28,30 Several biological and physical-chemical aspects of Zinc were recently elucidated in a multi-center study, [29][30][31][32][33] including effects on macrophage activity in vitro, such as fine modulation of hydrogen peroxide and cytokine synthesis. 20,33 In turn, Sil was associated with increased production of platelet-activating factor by activated macrophages. 34 Curiously, and of note here, an inverse relation between the incidence of silicosis and tuberculosis was seen in Zambia, after a retrospective epidemiological analysis. ...
... Macrophages were proven to be sensitive to the effects of high dilutions. [18][19][20][21][22][23] In the present study, we describe several aspects of specific and non-specific effects of Silicea terra and Zincum metallicum on BCG-challenged RAW 264.7 macrophages. ...
Silica (silicon dioxide) is found in nature, being composed of 2 chemical elements abundant on the Earth's crust, oxygen and silicon. As homeopathic medicine, silica is used for treatment of chronic ulcers due to its ability to modulate the macrophage activity. Zinc is a cofactor of several immune mediators, especially thymulin, which is also capable of modulating the macrophage activity and recruitment of B1 cells in mice. In the present study, we assessed the homeopathic medicines Silicea terra and Zincum in an in vitro experimental model to determine their effects on the interaction between RAW 264.7 macrophages and BCG (Bacillus Calmette-Guérin). In this step of the study, we assessed oxidative activity through measurement of hydrogen peroxide and nitrite/nitrate in the cell culture supernatant after 24 hours of treatment. Several homeopathic potencies were used for both drugs (6cH, 30cH, 200cH). The tests were performed in duplicate, and the data were analyzed by means one-way ANOVA. The results show no effect on nitric oxide (NO) production, but reduction of hydrogen peroxide production (p≤ 0.001) after treatment with the vehicle (0.03% alcohol). Such reduction was reversed with treatments Silicea terra30cH and Zincum metallicum 30cH (p≤ 0.001). Treatment with Silicea terra 200cH induced significant reduction of hydrogen peroxide production, even when compared to the vehicle (p≤ 0.005). Taken together, the results indicate that only treatment of BCG-challenged macrophages with Silicea terra 200cH was able to significantly reduce the oxidative activity of these cells after 24 hours of incubation. Since NO production in vitro usually occurs after 96 to 120 hours of incubation, one might infer that the negative result obtained in the present study might be associated with the time-point of assessment. Other aspects of the macrophage-BCG interaction are still under evaluation.
... 28,30 Several biological and physical-chemical aspects of Zinc were recently elucidated in a multi-center study, [29][30][31][32][33] including effects on macrophage activity in vitro, such as fine modulation of hydrogen peroxide and cytokine synthesis. 20,33 In turn, Sil was associated with increased production of platelet-activating factor by activated macrophages. 34 Curiously, and of note here, an inverse relation between the incidence of silicosis and tuberculosis was seen in Zambia, after a retrospective epidemiological analysis. ...
... Macrophages were proven to be sensitive to the effects of high dilutions. [18][19][20][21][22][23] In the present study, we describe several aspects of specific and non-specific effects of Silicea terra and Zincum metallicum on BCG-challenged RAW 264.7 macrophages. ...
Background The homeopathic medicines Silicea terra (Sil) and Zincum metallicum (Zinc) modulate macrophage activity and were assessed in an experimental study in-vitro for their effects on macrophage–BCG (Bacillus Calmette–Guérin) interaction.
Methods RAW 264.7 macrophages were infected with BCG, treated with different potencies of Sil and Zinc (6cH, 30cH and 200cH) or vehicle, and assessed 24 and 48 h later for bacilli internalization, hydrogen peroxide (H2O2) and cytokine production, and lysosomal activity.
Results Treatment with vehicle was associated with non-specific inhibition of H2O2 production to the levels exhibited by uninfected macrophages. Sil 200cH induced significant reduction of H2O2 production (p < 0.001) compared with the vehicle and all other treatments, as well as higher lysosomal activity (p ≤ 0.001) and increased IL-10 production (p ≤ 0.05). Such effects were considered specific for this remedy and potency. The number of internalized bacilli was inversely proportional to Zinc potencies, with statistically significant interaction between dilution and treatment (p = 0.003). Such linear-like behavior was not observed for Sil dilutions: peak internalization occurred with the 30cH dilution, accompanied by cellular degeneration, and IL-6 and IL-10 increased (p ≤ 0.05) only in the cells treated with Sil 6cH.
Conclusion Sil and Zinc presented different patterns of potency-dependent effect on macrophage activity. Bacterial digestion and a balanced IL-6/IL-10 production were related to Sil 6cH, though reduced oxidative stress with increased lysosomal activity was related to Sil 200cH. Degenerative effects were exclusively related to Sil 30cH, and potency-dependent phagocytosis was related only to Zinc.
... The literature to date already shows that homeopathic medicines do modulate macrophage activation patterns. 149,150 As a result, an area worthy of future study is the nature of the protein coronas for people with different homeopathic constitutional types or acute health conditions at baseline and over the course of treatment. This type of emergent biological identity from protein coronas potentially distinguishes homeopathic treatments using potentization-induced nano-structures from the non-individualized chemical identity of conventional drugs or even non-homeopathic nanostructures from outside the body. ...
... 158,170 A feature of the biological response is modulation of complex gene expression patterns by specific medicines. 150,[186][187][188] These adaptations are all in the service of optimizing the individual's fitness to survive in a biologically perceived environment that includes the potential disease damage or disease-encapsulated "danger" of the homeopathic medicine, as signaled by detection of the low-dose homeopathic information (that is, an individualized integrated form of the current disease state of the person). ...
Background:
Evidence indicates that homeopathic medicines are complex self-organizing nano-scale systems that generate unique low-intensity electromagnetic signals and/or quantum coherence domains. In Part 1, we reviewed relevant concepts from complex adaptive systems science on living systems for the nature of homeopathic healing.
Aim:
In Part 2, we discuss the complex-system nature of homeopathic medicines. The aim is to relate the evidence on the nature and properties of homeopathic medicines to the complex systems model for homeopathic healing.
Methods and results:
The work is a narrative review, with complexity model development for the nature of homeopathic medicines. Studies suggest that homeopathic manufacturing generates nano-structures of source material, silica and silicon quantum dots if succussed in glassware or including botanical source materials; or carbon quantum dots if succussed in plastic or including any organic source materials, as well as solute-induced water nano-structures carrying medicine-specific information. On contact with physiological fluids (e.g., blood plasma), there is evidence that nano-structures additionally adsorb individualized patterns of the recipient's own proteins on to their surfaces to create a unique protein corona coat (shell). Thus, the simillimum may generate a personalized biological identity upon administration. Consequently, a medicine can serve as an individually salient, self-similar information carrier, whose protein corona constituent pattern reflects the individual's current internal state of health/disease. Homeopathic medicine complexity emerges from interactions of the component parts from source, silica from glassware or carbon from plastic containers, solvents (lactose, water, ethanol), adsorbed biomolecule layers from plant or animal sources, and adsorbed biomolecules of the recipient. Low doses of these complex medicines can act as biological signaling agents to initiate hormesis via a network-wide pattern of adaptive responses by the recipient complex adaptive system, rather than as conventional pharmaceutical drugs. Biological mediators of adaptive responses include inter-connected network elements of the cell danger/damage defense system: for example, gene expression, reactive oxygen species, heat shock proteins, cytokines, macrophages, T-cells, and associated brain-immune system mediator pathways.
Conclusions:
Every homeopathic medicine is a complex nano-scale system involving multiple inter-connected, interacting components, and emergent properties. Simillimum individualization derives from formation of a unique personalized protein corona shell adsorbed to the reactive surface of the homeopathic nano-structures on contact with the recipient's body fluids. Low doses of such complex nano-structures initiate the adaptive processes of hormesis to mobilize endogenous healing of a disease state. The capacity for self-organization and self-similarity in complex systems is the key to future research on the nature of homeopathic medicines and systemic healing during individualized homeopathic treatment.
... This is in agreement with the view of Bellavite et al. [64,67] who claimed that the specificity of any drug is based on the interaction of active principles with their biological targets (enzymes, cell receptors, membrane channels, transcription factors in gene expression), and that the same is conceivably true for homeopathic drugs. These authors could show changes in the expression of key genes, mainly cytokines, receptors and transcription factors, induced by the homeopathic remedies Gelsemium sempervirens and Arnica Montana, likely to understand their anxiolytic, anti-inflammatory and wound-healing bio-effects, observed in clinical practice [102,103]. Significant effects were found essentially with the lowest dilutions (C2-C3). On the contrary, some paradoxical inverse effects were observed at high dilution (C15-C30); this will be discussed below. ...
... In the study of arsenic D45 on wheat germination reported above [125], an inhibitory effect was observed for 8 and 16 succussions, and a stimulating effect for more than 32 succussions; strikingly, this threshold corresponded to the onset of well-organized polycrystalline structures observed in the dilutions. In the study of Gelsemium and Arnica [102,103], inverse effects in the expression of genes were unexpectedly found at high C15 and C30 dilutions, compared to the normal, expected effects at low dilution. Chikramane et al. [86] using MTT, recently reported hormetic effects by homeopathically diluting-by successive factors of 1/3 and 1/10-high dilutions (C6, C30, C200) of metal NPs as starting solutions. ...
... Starting from year 2011, our publications have been the subject of several critical annotations by Dr. Salvatore Chirumbolo [27][28][29][30], and a summary of our technical responses was reported immediately afterward [31]. More recently, we used real-time polymerase chain reaction to investigate the effects of Arnica montana on gene expression of the THP-1 myelomonocytic cell line, differentiated by phorbol-myristate acetate and interleukin-4 in the woundhealing phenotype [32]. These findings also drew critical commentary by Chirumbolo and Bjørklund [33], based on some recalculations and extrapolations from the values of standard errors of the mean. ...
... Increase in gene expression of some CXC chemokines and bone morphogenetic protein, decrease in MMP-1 [32] A. montana 2-15 c (nonlinearity) Cultivated macrophages differentiated by IL-4 ...
Arnica montana is a popular traditional remedy widely used in complementary and alternative medicine, in part for its wound-healing properties. The authors recently showed that this plant extract and several of its homeopathic dilutions are able to modify the expression of a series of genes involved in inflammation and connective tissue regeneration. Their studies opened a debate, including criticisms to the “errors” in the methods used and the “confounders and biases”. Here the authors show that the criticisms raised on methodology and statistics are not consistent and cannot be considered pertinent. The present comment also updates and reviews information concerning the action of A. montana dilutions in human macrophage cells while summarizing the major experimental advances reported on this interesting medicinal plant.
... Additionally, homeopathy was one of the therapies most commonly sought for rheumatologic diseases such as progressive destruction and disability of joints, and as a patient preference due to intolerance of adverse effects with conventional treatment. Individualised homeopathic remedies have also been shown to produce positive effects for rheumatologic conditions [71]. In addition, an investigation into the effects of homoeopathic treatment of children suggested significant improvement of symptoms of upper respiratory tract infections. ...
Homeopathy was founded some two hundred years ago by Dr Samuel Christian Hahnemann. Over time, it has grown to be among the most frequently used forms of alternative medicine in Europe and the USA. It is underpinned by the principle of ‘like cures like’, where highly diluted substances are used for therapeutic purposes, by producing similar symptoms to when the substance is used in healthy people. Many studies have been published on the value of homeopathy in treating diseases such as cancer, depression, psoriasis, allergic rhinitis, asthma, otitis, migraine, neuroses, allergies, joint disease, insomnia, sinusitis, urinary tract infections and acne, to name a few. We conducted a comprehensive review of the literature on homeopathy and evaluated its effectiveness in clinical practice. While there is evidence of the clinical benefits of homeopathy, its formal application requires more rigorous randomised controlled trials.
... It leads to reduced expression of the cell surface receptors CD25, CD28, CD27 and CD120b, which play a key role in the activation of NF-kB in T cells. This supports the mechanism proposed by Lyss in 1997 [48][49]. NF-kB activation is associated with the induction of pain and inflammation, characterized by the release of proinflammatory cytokines (tumor necrosis factoralpha [TNF-a] and interleukin-1beta [IL-1b] and local leukocyte recruitment [50]. ...
Background: Despite scientific advances, the management of procedural pain in neonates remains suboptimal. Applying adequate therapy to control pain during the neonatal period is a moral and ethical act. In recent decades, ample evidence has accumulated regarding the risks, associated with both untreated pain and the use of more aggressive analgesic therapy. Thus, the emphasis in neonatal clinical practice is on non-aggressive pain management techniques, including homeopathic agents. The efficacy and safety of homeopathic agents for reducing procedural pain in neonates is the subject of the present study. Materials and Methods: Healthy full-term newborn babies with an average age of 72-84 hours were included. They were divided into three study groups: group (A) – not anesthetised, /n= 67/, group (B) – anesthetised with Arnica D30 /n=40/ and group (C)- anesthetised with Sol.Glucose 25% /n=40/. The severity of the pain was assessed using the multimodal - Neonatal Infant Pain Scale (NIPS) and the unimodal - Neonatal Facial Coding System (NFCS) scale. Assessments were done 30s before, 30 seconds after, and 5 minutes after the painful stimulus caused by the heel prick. Heart rate, transcutaneous oxygen saturation, respiratory rate and blood pressure were examined at the same intervals. The results obtained were processed statistically by descriptive analysis and ANOVA at a significance level of p< 0.05. Results: 5 minutes after heel prick both scales showed a statistically significant lower score in those given Arnica D30, compared to those who received Sol.Glucose 25%. When monitoring the physiological indicators, we found a significant increase in the heart rate 30 seconds after the procedure.5 minutes after the heel prick we recorded significantly respiratory rate and systolic blood pressure in all three groups compared to pre-procedural values. Conclusions: Arnica D30 has better analgesic effects compared to Sol.Glucose 25% for newborn babies after heel prick tests.
... It leads to reduced expression of the cell surface receptors CD25, CD28, CD27 and CD120b, which play a key role in the activation of NF-kB in T cells. This supports the mechanism proposed by Lyss in 1997 (Iannitti et al. 2016;Olioso et al. 2016). NF-kB activation is associated with the induction of pain and inflammation, characterized by the release of proinflammatory cytokines (tumor necrosis factor-alpha [TNF-a] and interleu-kin-1beta [IL-1b] and local leukocyte recruitment (Kawakami et al. 2011). ...
Background: Despite scientific advances, the management of procedural pain in neonates remains suboptimal. Applying adequate therapy to control pain during the neonatal period is a moral and ethical act. In recent decades, ample evidence has accumulated regarding the risks, associated with both untreated pain and the use of more aggressive analgesic therapy. Thus, the emphasis in neonatal clinical practice is on non-aggressive pain management techniques, including non-pharmacological methods, such as glucose and homeopathic agents. The efficacy and safety of homeopathic agents for reducing procedural pain in neonates is the subject of the present study.
Materials and methods: Healthy full-term newborn babies with an average age of 72–84 hours were included. They were divided into three study groups: group (A) – without analgesia, (n = 67), group (B) – analgesic with Arnica D30 (n = 57) and group (C) – analgesic with Sol.Glucose 25% (n = 40). The severity of the pain was assessed using the multimodal - Neonatal Infant Pain Scale (NIPS) and the unimodal - Neonatal Facial Coding System (NFCS) scale. Assessments were done 30s before, 30 seconds after, and 5 minutes after the painful stimulus caused by the heel prick. Heart rate, transcutaneous oxygen saturation, respiratory rate and blood pressure were examined at the same intervals. The results obtained were processed statistically by descriptive analysis and ANOVA at a significance level of p < 0.05.
Results: Five minutes after heel prick both scales showed a score near 3, i.e. no significant pain, and statistically significant lower score in those given Arnica D30, compared to those who received Sol.Glucose 25% (p < 0.05). When monitoring the physiological indicators, we found a significant increase in the heart rate 30 seconds after the procedure between group B and C. Five minutes after the heel prick we recorded significantly values of respiratory rate and systolic blood pressure in all three groups compared to accounted for pre-procedural values.
Conclusion: Arnica D30 has better analgesic effects compared to Sol.Glucose 25% for newborn babies after heel prick tests.
... Olioso i wsp. [13] zwracają uwagę na wynikające z ich badań reakcji łańcuchowej polimerazy nowe hipotezy dotyczące wpływu tej rośliny na ludzkie makrofagi. Co ważne, wykazano skuteczność maści na bazie arniki górskiej w leczeniu procesu zapalnego i uszkodzeń oksydacyjnych wywołanych promieniowaniem UVB. ...
Jednym z najczęstszych powodów zgłaszania się pacjentek do fizjoterapeuty uroginekologicznego jest blizna po operacji ginekologicznej (np. usunięcia macicy), uroginekologicznej (np. leczenia zaburzenia statyki narządów miednicy mniejszej lub nietrzymania moczu) lub położniczej (np. cięcia cesarskiego). Konsekwencją przebytej operacji mogą być: blizna przerostowa lub bliznowiec, przewlekły ból miednicy, dyspareunia, niepłodność, nieswoiste dolegliwości układów pokarmowego, moczowego lub rozrodczego, zespoły bólowe narządu ruchu lub zaburzenie wzorca postawy ciała i chodu. Aby przeciwdziałać konsekwencjom zrostów pooperacyjnych lub je ograniczyć, należy w sposób planowany wdrożyć terapię blizny, najlepiej z wykorzystaniem odpowiednich środków farmakologicznych jako podłoża. W przypadku wielu z przeznaczonych do tego celu preparatów nie udowodniono ich skuteczności lub jej nie badano. W artykule omówiono substancje aktywne wchodzące w skład preparatów do leczenia blizn, wyszczególniając ich charakterystykę, mechanizm działania, aspekty farmakologiczne i dostępne badania naukowe na ich temat. Przedstawiono również autorski plan terapii blizny, uwzględniający etapy gojenia się rany, i wskazano metody, w których zasadne jest zastosowanie środków farmakologicznych.
... Some groups serially diluted the ethanolic extract of the medicines and gave 20 succussions during preparation. 5 Other groups gave 10 succussions for their experiments after dilution. 6 In another instance, the experiments were conducted by succussing the medicines just prior to addition to the cells. ...
Standardization of dosing in an in vitro model.
... Arnica montana is used topically to treat some ailments such as joint pain, bruises, wound healing, muscle aches, sprains, superficial phlebitis, insect bite swelling, and inflammation from bone breaks. Recent research indicates that it might be beneficial in treating scalds [68,69]. Increased total phenolic acid levels in Arnica montana L. roots were also correlated with mycorrhization [70]. A. montana contains 150 therapeutically active substances, which include sesquiterpene lactones such as helenalin and 11 alpha, 13-dihydohelenalin, together with their carbonic acid ester short chains (0.1-0.5% of dry matter in leaves, 0.3-1% in flower shoots), flavonoids (0.6-1.7%) using capillary electrophoresis [71] in the form of flavonoid glucuronides, flavonoid glycosides, and flavonoid aglycones. ...
... Together, these findings suggest the existence of two distinct cell populations whereby only one would be able to respond to Aspirin 15cH-possibly due to epigenetic or phenotypic modulation, since this kind of effect has previously been described in other cell models for studying the action of homeopathic formulations on macrophages in vitro. [45][46][47] Besides F-actin, dynamic alteration of macrophage morphology depends on the rapid rearrangement of cytoskeletal microtubules in response to extracellular stimuli. 48 Bian et al demonstrated that stimulation of TLR-4 by LPS reduces the expression of αand β-tubulins, increasing microtubule plasticity and allowing changes in cell morphology. ...
Introduction Aspirin is one of the most commonly used drugs worldwide. It is known to present antipyretic, anti-inflammatory and anti-thrombotic actions, making it extremely useful in a wide range of clinical contexts. Interestingly, homeopathically prepared Aspirin 15cH has been found to have a pro-thrombotic effect in rats, raising the hypothesis that Aspirin 15cH could also modulate the activity of inflammatory cells in different pathological processes.
Objective Our objective was to assess what effect Aspirin 15cH has on RAW 264.7 macrophages in vitro.
Methods The effects of Aspirin 15cH on biochemical and morphological activities of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages were evaluated. These effects were compared with unchallenged macrophages (negative control), untreated LPS-stimulated macrophages, macrophages treated with succussed water (vehicle control), or aspirin 200 µg/mL (pharmacological inhibitor of LPS activity). Cell morphology (adhered cell area and cytoskeleton arrangements), cell viability, toll-like receptor-4 (TLR-4) expression, and the production of nitric oxide, cytokines and intracellular reactive oxygen species were assessed.
Results Aspirin 15cH reduced the number of cells expressing TLR-4 on the surface (p = 0.03) and induced a “columnar” morphology of macrophage pseudopods, indicating changes in cytoskeleton arrangement. When cells were treated with both Aspirin 15cH and LPS, cell morphology became heterogeneous, suggesting that sub-populations of cells had differing sensitivities to LPS or Aspirin 15cH. Exposure of the cells to LPS alone, succussed water or aspirin 200 µg/mL produced effects consistent with the literature.
Conclusion Aspirin 15cH, aspirin 200 µg/mL, LPS and succussed water appear to act as independent stimuli able to induce different patterns of macrophage response. Aspirin 15cH induced changes suggestive of M2 polarization of the macrophages (i.e., toward a wound healing or tissue repair, rather than inflammatory, phenotype). These preliminary findings need to be confirmed in further specific studies.
... Arsenicum album (30C and 200C) has been reported against Arsenate induced oxidative stress [13,14]. Different homeopathic medicines like Arnica montana, 30C, 200C [15,16], Ruta graveolens 30C, 200C [17], Ginseng 30C, 200C, and Hypericum 200C, against shock and injury associated with X-rays [18,19]. Many plantbased homeopathic medicines mentioned in the repertory have been reported to manage diabetic mellitus [20]. ...
Background:
Plant-derived homeopathic medicines (HMs) are cheap and commercially available but are mechanistically less explored entities than conventional medicines.
Purpose:
The aim of our study was to evaluate the impact of selected plant-derived HMs derived from Berberis aquifolium (BA), Berberis vulgaris (BV), Mentha piperita (MP), Curcuma longa (CL), Cinchona officinalis (CO), Thuja occidentalis (TO) and Hydrastis canadensis (HC) on cervical cancer (CaCx) cells in vitro.
Methods:
We screened the mother tincture (MT) and 30C potencies of the above-mentioned HMs for anti-proliferative and cytotoxic activity on human papillomavirus (HPV)-negative (C33a) and HPV-positive CaCx cells (SiHa and HeLa) by MTT assay. Total phenolic content (TPC) and the free-radical scavenging activity of each HM was also determined using standard assays. Phytochemicals reportedly available in these HMs were examined for their potential inhibitory action on HPV16 E6 by in silico molecular docking.
Results:
All tested MTs induced a differential dose-dependent cytotoxic response that varied with cell line. For C33a cells, the order of response was TO > CL > BA > BV > HC > MP > CO, whereas for SiHa and HeLa cells the order was HC > MP > TO > CO > BA > BV > CL and CL > BA > CO, respectively. 30C potencies of all HMs showed an inconsistent response. Further, anti-CaCx responses displayed by MTs did not follow the order of an HM's phenolic content or free radical scavenging activity. Analysis revealed anti-oxidant content of BA, BV and HC had the lowest contribution to their anti-CaCx activity. Using in silico modeling of molecular docking between the HPV16 E6 protein crystallographic structures (6SJA and 4XR8) and main phytochemical components of BV, BA, HC, CL and TO, their potential to inhibit the HPV16 E6 protein carcinogenic interactions was identified.
Conclusion:
The study has shown a comparative evaluation of the potential of several plant-derived MTs and HMs to affect CaCx cell line survival in vitro (through cytotoxicity and free radical scavenging) and their theoretical molecular targets in silico for the first time. Data demonstrated that MTs of BA and BV are likely to be the most potent HMs that strongly inhibited CaCx growth and have a strong anti-HPV phytochemical constitution.
... Arsenicum album (30C and 200C) has been reported against Arsenate induced oxidative stress [13,14]. Different homeopathic medicines like Arnica montana, 30C, 200C [15,16], Ruta graveolens 30C, 200C [17], Ginseng 30C, 200C, and Hypericum 200C, against shock and injury associated with X-rays [18,19]. Many plantbased homeopathic medicines mentioned in the repertory have been reported to manage diabetic mellitus [20]. ...
Objectives
Antioxidant property like radical scavenging is a primary target to elucidate the efficacy mechanism of a drug against diseases linked to oxidative stress such as cancer, metabolic disorders, rheumatoid arthritis, etc. In alternative therapies, homeopathy is one of the preferred choices by patients and clinicians due to its potential to cure chronic and complex illnesses. However, the efficacy of homeopathic preparations at high diluted potencies attracts rational criticism due to insufficient scientific knowledge supporting the mechanism of action. Therefore, an attempt was made to estimate the total phenolic content (TPC) and radical scavenging activity of clinically prescribed homeopathic drugs.
Methods
With gallic acid as a reference control, mother tinctures (MTs) and different potencies of Eucalyptus globulus (EG), Syzygium jambolanum (SJ), Ruta graveolens (RG), and Thuja occidentalis (TO) were used to perform Folin–Ciocalteu test, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays.
Results
The results showed TPC of MTs equivalent to µg/mL of gallic acid viz; EG (4,872.5 ± 133.2), SJ (8,840.5 ± 14.8), RG (985.6 ± 39.1), and TO (341.5 ± 19.5) with significant ABTS and DPPH radical scavenging potential. Whereas 30C and 200C potencies of each homeopathic drug showed undetectable phenolic content and insignificant radical scavenging potential compared to vehicle control, i.e., alcohol 90% (2.0 ± 1.5).
Conclusions
The reported efficacy of 30C and 200C potencies of homeopathic medicines against oxidative stress-related illnesses might be due to mechanisms other than radical scavenging. Furthermore, the assays studied can be helpful in drug standardization and quality control of MTs that are used as starting material in homeopathic preparations.
... Arnica montana is a protected plant species used widely in traditional and modern medicine for its antibacterial, antitumour, antioxidant, anti-inflammatory, and antifungal properties (Knuesel et al., 2002;Galambosi, 2004;Macêdo et al., 2004;Gawlik-Dziki et al., 2011;Craciunescu et al., 2012;Kowalski et al., 2015;Olioso et al., 2016;Kriplani et al., 2017;Sugier et al., 2019Sugier et al., , 2020. The genus has more than 30 different species, and they differ significantly in the content of medicinal components, with A. montana being the species used most widely for its medicinal value (Kowalski et al., 2015;Kriplani et al., 2017) and the flower heads being the part of the plant containing the highest levels of biologically active compounds (Galambosi, 2004;Ganzera et al., 2008;Spitaler et al., 2008). ...
Arnica montana L. flower heads are known for their antioxidant, antimicrobial, and anticancer activity. The aim of this work was to optimize the process of supercritical CO2 extraction, to achieve high extraction yield and high content of biologically active components, and to confirm the antimicrobial and anticancer activity of the extract. The influence of pressure and temperature on the total phenolic content, antioxidant activity, and proanthocyanidin content was evaluated. The pressure and temperature were found to be interdependent. A temperature of 60°C and a pressure of 30 MPa resulted in a high extraction yield, antioxidant activity and phenolic content. The content of proanthocyanidins was highest at a pressure between 18 and 24 MPa. The extracts inhibited three different microorganisms successfully; Staphylococcus aureus, Escherichia coli and Candida albicans, at concentrations ranging from 0.1 to 5.16 mg/ml and showed anticancer activity decrease up to 85% at a concentration of 0.5 mg/ml.
... This study evaluates the use of a topical cream composed by natural extracts with complementary and synergistic effects for pain management. Menthol reduces pain and increases blood flow, warming up the muscle and enhancing absorption of the rest of the extracts [24][25][26][27][28]. Arnica montana reduces pain, has anti-inflammatory effects and potentiates tissue repair [29,30]. Hypericum perforatum provides anti-inflammatory effect and drives tissue regeneration and scarring [31][32][33][34]. ...
... A. montana extracts have also been reported to promote healing, as some of the modified genes are key factors of tissue remodeling, inflammation, and chemotaxis [35]. In addition, Mahajan et al. ...
... This study evaluates the use of a topical cream composed by natural extracts with complementary and synergistic effects for pain management. Menthol reduces pain and increases blood flow, warming up the muscle and enhancing absorption of the rest of the extracts [24][25][26][27][28]. Arnica montana reduces pain, has anti-inflammatory effects and potentiates tissue repair [29,30]. Hypericum perforatum provides anti-inflammatory effect and drives tissue regeneration and scarring [31][32][33][34]. ...
Introduction: Musculoskeletal pain is a common affection due to ageing, sedentarism and injuries. The objective of this trial is to prove efficacy of a natural topical composition containing Arnica montana, Hypericum perforatum, Calendula officinalis, Melaleuca sp. and menthol in pain management in adults with acute or chronic pain.
Methods: This randomized, double-blinded and placebo-controlled trial included 200 patients with musculoskeletal pain, 100 in the intervention group receiving the topical formula and 100 in the placebo group, receiving a similar formula without active ingredients. The products were applied topically twice daily for 14 days in affected areas. Immediate pain alleviation and stiffness perception were monitored for two hours at days 0, 7 at 14. Pain reduction and recovery perception upon sustained application were assessed after 7 and 14 days.
Results: Intervention immediately reduced pain and stiffness at rest and in motion 30 minutes after application and kept being superior to placebo in all short-term timepoints (p < 0.05). Immediate pain reduction was maintained even at late stages of recovery. A two-week sustained intervention resulted in significant pain reduction and improvement in recovery perception. Even if both groups reached statistical significance with respect to baseline due to spontaneous lesion recovery, a significantly improved recovery was reported in the intervention group with respect to placebo.
Conclusions: Intervention was found to reduce pain and stiffness upon minutes of its application and to improve pain and mobility over the 14 days of treatment, showing benefits both for immediate alleviation and for longer term recovery.
... The cells differentiated with Interleukin-4 induced a "wound-healing"-like phenotype and were then treated with various homeopathic dilutions of Arnica montana (2C, 3C, 5C, 9C, and 15C) for 24 hours. The expression of several genes of cytokines [66] and extracellular matrix [52,53] was modulated. Another interesting observation was that on subsequent dilution, the activity of most genes whose expression was changed by Arnica montana 2C remained modified in the same direction (i.e., upregulated genes remained upregulated and downregulated remained downregulated) even at higher dilutions. ...
Homeopathy is often attacked with claims of not being evidence-based or for the implausible nature of its major principles of dilution/potentization and/or the similars (“similia similibus curentur”). However, these statements have already been falsified on the experimental ground, besides being incorrect on an epistemological level. Here we provide an updated appraisal of the scientific approach to the principle of Similars and homeopathy, focusing on laboratory models. After a brief historical introduction concerning the early scientific investigations, some recent in vitro studies are reviewed, with particular reference to those from our group. The second part is devoted to explaining the homeopathic principle of Similars using conceptual models in the field of complexity science and the theory of dynamic systems.
... Estes resultados corroboram com o trabalho de Castro et al. (2012) que analisaram o efeito da A. montana e sua associação com microcorrente em feridas no dorso de ratos. Outros estudos mostraram a ação desta espécie na angiogênese em feridas de pele no dorso de ratos (OLIOSO et al., 2016;MEDRADO et al., 2003). ...
A cicatrização de feridas é um problema de saúde pública e, por isso, o processo de reparo tecidual tem merecido atenção de pesquisadores, principalmente no que se refere aos fatores que o retardam ou dificultam. A aplicação de microcorrente em lesões teciduais desempenha papel fundamental no processo de cicatrização sendo capaz de restaurar o fluxo elétrico na região da lesão, além de contribuir para diminuição da contaminação bacteriana. Também a utilização de plantas medicinais como Arnica montana é de grande interesse, pois esta espécie possui a propriedade de acelerar o processo de cicatrização quando utilizada topicamente sobre a região afetada. Neste sentido, o presente estudo visa analisar os efeitos da estimulação com microcorrente associada ou não à aplicação tópica de A. montana em lesões incisionais na pele de ratos. Trinta e seis animais, de 90 dias, com 250g, foram divididos aleatoriamente em (n=3): (C) sem tratamento; (MC) tratado com microcorrente (10 µA/2 min); (A) tratado com aplicação tópica de gel contendo A. montana; (A+MC) tratado com aplicação tópica de gel contendo A. montana + microcorrente (10 µA/2 min).. Amostras foram coletadas dos animais eutanasiados no 2º, 6º e 10º dia após a lesão cirúrgica para análise histomorfométrica: quantificação de infiltrado inflamatório, vasos sanguíneos, fibroblastos e porcentagem da área de colágeno. Para análise estatística foi utilizado ANOVA Two-way e pós-teste de Tukey (média±erro padrão). As terapias combinadas ou não, diminuíram a quantidade de infiltrado inflamatório e aumentaram o número de fibroblastos, colágeno e vasos sanguíneos durante o período experimental. A associação da aplicação de microcorrente e A. montana foi efetiva no reparo tecidual de lesões incisionais na pele do dorso de ratos, controlando a inflamação, favorecendo a fibroplasia, colagênese e angiogênese.
... Le principe actif, sous forme moléculaire, assurerait une action spécifique au niveau des cibles de la pharmacologie classique (enzymes, récepteurs cellulaires, canaux membranaires, facteurs de transcription). Bellavite et coll [54,55] ...
Les hautes dilutions homéopathiques vues par la physique : arguments en faveur d'une nanomédecine Homeopathic high dilutions seen by physics argue for a nanomedicine Jean-Louis Demangeat (ancien chef de service de médecine nucléaire du Centre hospitalier de Haguenau et ancien maître de conférences en biophysique à la Faculté de médecine de Strasbourg) 13d, rue d'Oberhausbergen, RÉSUMÉ La critique de l'homéopathie repose sur l'absence théorique de l'ingrédient de départ dans les dilutions ultramoléculaires au-delà de 12 CH (limite d'Avogadro), et corrélativement sur l'absence de mécanisme d'action biologique. Or, la préparation spécifique par dilution/dynamisation implique des processus d'interaction itératifs avec l'atmosphère (nanobulles), les parois des contenants (silice et autres éléments chimiques), et avec la micropipette de prélèvement (adsorption). La RMN a montré depuis 2003 des nanostructures (NS) dans les dilutions ultramoléculaires, confirmées plus récem-ment par diverses techniques (filtration, diffusion laser, diffraction RX, microscopie électronique). Ces nanostructures/nanoparticules (NS/NPs) persistent et même s'accroissent avec la hauteur de dilution, et sont différenciables selon la substance de départ, à condition d'avoir été dynamisées. Plusieurs études montrent la présence inattendue de matériel dans les hautes dilutions, et souvent même la rémanence du matériel initial au-delà de 12 CH. Ces NS/NPs ne sont pas observées à basse dilution. La gamme 2-4 CH correspond à la transition entre l'état « moléculaire » et « nanoparticulaire » du principe actif qui pourrait expliquer les inversions d'effets observés en biologie. Compte tenu des propriétés particulières de biodisponibilité et réactivité des NPs, l'hypo-thèse des hautes dilutions agissant comme nanomédecine peut raisonnablement être avancée. © 2020 Elsevier Masson SAS. Tous droits réservés. SUMMARY The criticism of homeopathy is based on the theoretical absence of the starting ingredient in ultramolecular dilutions above 12 CH (Avogadro limit), and correlatively on the absence of a biological mechanism of action. However, the specific preparation by dilution/dynamisation involves iterative interaction processes with the atmosphere (nanobubbles), the walls of the containers (silica and other chemical elements), and with the sampling micropipette (adsorption). Since 2003, NMR has shown nanostructures (NS) in ultramolecular dilutions, confirmed more recently by various techniques (filtration, laser diffusion, X-ray diffraction, electron microscopy). These nanostructures/nanoparticles (NS/NPs) persist and even increase with the dilution level, and can be distinguished according to the starting substance, provided they have been dynamized. Several studies show the unexpected presence of material in high dilutions, and even the persistence of the initial material beyond 12 CH. These NS/NPs are not observed at low dilution. The 2-4 CH range corresponds to the transition between the "molecular'' and the "nanoparticulate'' state of the solute, which could explain inversion effects observed in biology. Given the particular bioavailability and reactivity properties of NPs, the hypothesis of high dilutions acting as nanomedicine can reasonably be postulated.
... Arnica montanaAnti-inflammatory, Antioxidant, and Helenalin (sesquiterpene lactone),(Olioso et al., 2016; Craciunescu et cytoprotective flavonoids, and phenolic acids al., 2012;Lyß et al., 1998) Atropa belladonna Sedative, local analgesic, hypertensive Atropine (alkaloid), scopolamine(Bousta et al., 2001; Kwakye et al.Rakelly de Oliveira et al., 2015; marianum L. Gaertn inflammatory, antitumor, antibacterial, flavonolignans): major components Ebrahimpour koujan et al., 2015;are silybin, taxifolin, and quercetinWellington and Jarvis, 2001) (flavonoids)ChenopodiumCytotoxic against tumor cell linesAscaridol isolated from essential(Yadav et al., 2007; anthelminticum Linnoils Efferth et al., 2002) Cinnamomum Antidiabetic, antioxidant, antimutagenic, Essential oils, such as eugenol and (Cardoso-Ugarte et al., 2016; zeylanicum antitumor, and antimicrobial cinnamaldehyde, and cynammic Husain et al., 2018; acetate Verspohl et al., 2005) Colchicum autumnale Antirheumatic and anti-inflammatory Colchicine (alkaloid) (Akram, 2012; Kurek, 2018) Echinacea Anti-inflammatory, immunostimulator, Flavonoids, lipophilic compounds, (Shaffique et al., 2018; Bonomo et angustifolia antioxidant and antimicrobial such as alkamides and ketoalkenes, al., 2017; Kindscher, 2016) and hydrophilic phenolic compounds, such as caffeic acid derivatives TABLE 8.ContinuedSánchez-Tena et al., 2012; Touriño virginiana antiviral, and inhibits cancer cell proliferation dins, hamamelitannin pentagalloyl-et al., 2008; Theisen et al., 2014) glucose, gallotannins, and gallates Juglans regia Cytotoxic against various human cancer cell Juglone (Panth et al., 2016) lines: lung (NCI-H322 and A549), breast (T47D), skin (A-431), prostate colon (Colo-205 and HCT-116), and prostate (PC- ...
... Le principe actif, sous forme moléculaire, assurerait une action spécifique au niveau des cibles de la pharmacologie classique (enzymes, récepteurs cellulaires, canaux membranaires, facteurs de transcription). Bellavite et coll [54,55] ...
Résumé
La critique de l’homéopathie repose sur l’absence théorique de l’ingrédient de départ dans les dilutions ultramoléculaires au-delà de 12 CH (limite d’Avogadro), et corrélativement sur l’absence de mécanisme d’action biologique. Or, la préparation spécifique par dilution/dynamisation implique des processus d’interaction itératifs avec l’atmosphère (nanobulles), les parois des contenants (silice et autres éléments chimiques), et avec la micropipette de prélèvement (adsorption). La RMN a montré depuis 2003 des nanostructures (NS) dans les dilutions ultramoléculaires, confirmées plus récemment par diverses techniques (filtration, diffusion laser, diffraction RX, microscopie électronique). Ces nanostructures/nanoparticules (NS/NPs) persistent et même s’accroissent avec la hauteur de dilution, et sont différenciables selon la substance de départ, à condition d’avoir été dynamisées. Plusieurs études montrent la présence inattendue de matériel dans les hautes dilutions, et souvent même la rémanence du matériel initial au-delà de 12 CH. Ces NS/NPs ne sont pas observées à basse dilution. La gamme 2-4 CH correspond à la transition entre l’état « moléculaire » et « nanoparticulaire » du principe actif qui pourrait expliquer les inversions d’effets observés en biologie. Compte tenu des propriétés particulières de biodisponibilité et réactivité des NPs, l’hypothèse des hautes dilutions agissant comme nanomédecine peut raisonnablement être avancée.
... The popular use of this species of arnica may be based on the proven use of mountain arnica, originating from the mountainous regions of Europe, which has been widely studied and is recommended for its anti-inflammatory action, although it is considered hepatotoxic and can be used in the homeopathic treatment, but not in herbal medicine. 17,18 The natural preference for the use of medicinal plants to fight diseases is embedded in everyday life. It is so natural and so easy to assimilate that those who use it feel appropriate to full knowledge. ...
Objetivo: Identificar as plantas utilizadas com fins terapêuticos por usuários de uma unidade pré-hospitalar pública do município de Campos dos Goytacazes/RJ, os modos de preparo e uso, e o conhecimento deles sobre às propriedades terapêuticas, mecanismos de ação e indicação das plantas que utilizam. Métodos: Pesquisa descritiva, exploratória e qualitativa, operacionalizada por dez entrevistas semiestruturadas em abril de 2016, cujos dados foram tratados por análise temática. Resultados: Foram citadas 40 plantas, sendo a mais frequente a erva cidreira (Lippia alba), utilizada como calmante. A principal forma de obtenção das plantas é por meio de cultivo próprio; e o chá, preparado por infusão ou decocção, a principal forma de consumo. Foram citados malefícios associados ao uso da arnica (Solidago chilensis). Conclusão: Verificou-se que a maior parte das plantas utilizadas é de procedência regional, cujo conhecimento de uso foi adquirido por mães e avós.
... This specific feature is also seen in other studies involving gene expression and homeopathy. 27,36,37 The acridine orange test showed an interesting result. While Ars 200cHP and Ars 200cHG induced a similar mor-phological pattern, characterized by patent acid vacuoles (lysosomes), Ars 200cHGC (superficial phase after centrifugation) induced a less active pattern. ...
Introduction According to the “silica hypothesis” formulated to explain homeopathy, the information of starting materials would be transferred to cells by silica nanoparticles detached from the glassware walls by serial dilution and agitation through epitaxy. We compared the biological activity, electrical current and silicon microparticle content (by means of scanning electron microscopy/energy-dispersive X-ray spectroscopy) of high dilutions (HDs) of arsenic prepared in plastic and glass vials to investigate the role of silica in their biological effects in vitro.
Materials and Methods Co-cultures of macrophages and yeast (Saccharomyces cerevisiae) were treated with different HDs of arsenic prepared in plastic and glass vials. Macrophage morphology, phagocytosis index, nitric oxide (NO), and cytokine production were evaluated.
Results Measurable amounts of silicon microparticles were detected only in the HDs prepared in glass vials, but ultra-centrifugation eliminated them. Specific and non-specific results were observed. Non-specific pro-inflammatory effects were seen in all dilutions prepared in plastic vials, including elevation of pro-inflammatory cytokines, NO and macrophage phagocytic index. Only the 200th centesimal dilution of arsenic produced specific decrease in interleukin-6 production in macrophages, and it was independent of the vial type or the presence of microparticles of silica in the medicine samples. The nature of the vials had an impact on the electric flow in the respective fluids.
Conclusion The non-specific, pro-inflammatory effects might be attributed to organic residuals detached from the vials' plastic walls during manipulation. Instead, specific silica-independent effects of the homeopathic medicine can be attributed to the decrease of interleukin-6 after treatment with the 200th centesimal dilution of arsenic.
... H omeopathic remedies (HRs) contain plant, animal, mineral, and metal products with specific odors such as flavonoids and terpenes in plant extracts, for example, [1][2][3][4][5] and two well-recognized homeopaths, Hahnemann and Schmidt, asked their patients to smell HRs. [6][7][8][9] Odorant molecules (OMs) and HRs share some properties: some OMs can enhance the perception of other OMs, and some odors can inhibit HRs efficiency. ...
Homeopathic remedies (HRs) contain odorant molecules such as flavonoids or terpenes and can lose their efficiency in presence of some competitive odors. Such similarities, along with extreme sensitivity of the olfactory system, widespread presence of olfactory receptors over all organic tissues (where they have metabolic roles besides perception of odors), and potential direct access to the brain through olfactory nerves (ONs) and trigeminal nerves, may suggest the olfactory system as target for HRs. Recent works highlighted that HRs exist in a dual form, that is, a still molecular form at low dilution and a nanoparticulate form at high dilution, and that remnants of source remedy persist in extremely high dilutions. From the literature, both odorants and nanoparticles (NPs) can enter the body through inhalation, digestive absorption, or through the skin, especially, NPs or viruses can directly reach the brain through axons of nerves. Assuming that HRs are recognized by olfactory receptors, their information could be transmitted to numerous tissues through receptor-ligand interaction, or to the brain by either activating the axon potential of ONs and trigeminal nerves or, in their nanoparticulate form, by translocating through axons of these nerves. Moreover, the nanoparticulate form may activate the immune system at multiple levels, induce systemic various biological responses through the pituitary axis and inflammation factors, or modulate gene expression at the cellular level. As immunity, inflammation, pituitary axis, and olfactory system are closely linked together, their permanent interaction triggered by olfactory receptors may thus ensure homeostasis.
... This situation could be characterised as a 'hormetic' effect. 41 Non-monotonic and hormesis-like responses, as well as oscillations, are indications of the highly non-linear and complex character of organisms and cellular systems, and very probably of the homeopathic phenomenon itself. 42 On closer examination of the similia principle itself, claimed by Vithoulkas as contrary to the whole concept behind Benveniste's experiment, 20 it is well known that several homeopathic remedies (Chamomilla, Arnica, Hypericum, to cite only a few) cure the same ailments or symptoms as those for which the mother tinctures (used in their preparation) are used. ...
We discuss questions related to the ‘Benveniste Affair’, its consequences and broader issues in an attempt to understand homeopathy. Specifically, we address the following points:
1. The relationship between the experiments conducted by Benveniste, Montagnier, their collaborators and groups that independently tested their results, to ‘traditional’ homeopathy.
2. Possible non-local components such as ‘generalised entanglement’ as the basis of the homeopathic phenomenon and experimental evidence for them.
3. The capability of highly diluted homeopathic remedies to provoke tangible biological changes in whole organisms and cellular experimental systems.
4. Aspects of the similia principle related to the above.
5. Suggestions that can lead to experimental verifications of the non-local hypothesis in homeopathy.
... Recent evidence from our laboratory suggests that Arnica montana stimulates the gene expression of chemokines and of connective tissue components by macrophages in vitro [70,71]; in our experiments the best effects required a pre-sensitization with interleukin-4 and were obtained with the low dilution 2c, but significant gene expression persisted also after treatment with 5c, 9c, and 15c A. montana. ...
Gelsemium sempervirens L. (Gelsemium) is traditionally used for its anxiolytic-like properties and its action mechanism in laboratory models are under scrutiny. Evidence from rodent models was reported suggesting the existence of a high sensitivity of central nervous system to anxiolytic power of Gelsemium extracts and Homeopathic dilutions. In vitro investigation of extremely low doses of this plant extract showed a modulation of gene expression of human neurocytes. These studies were criticized in a few commentaries, generated a debate in literature and were followed by further experimental studies from various laboratories. Toxic doses of Gelsemium cause neurological signs characterized by marked weakness and convulsions, while ultra-low doses or high Homeopathic dilutions counteract seizures induced by lithium and pilocarpine, decrease anxiety after stress and increases the anti-stress allopregnanolone hormone, through glycine receptors. Low (non-Homeopathic) doses of this plant or its alkaloids decrease neuropathic pain and c-Fos expression in mice brain and oxidative stress. Due to the complexity of the matter, several aspects deserve interpretation and the main controversial topics, with a focus on the issues of high dilution pharmacology, are discussed and clarified.
... This formulation containing Bryonia is reported to reduce oxidative metabolism in mouse macrophages. [41] A randomised controlled clinical study has concluded that the treatment of chronic low-back pain due to osteoarthritis using homoeopathic Bryonia has revealed that the combination of physiotherapy and homoeopathic drug treatment has benefi cial effects as compared to the placebo treatment. This study has stated the need for further investigations using larger sample sizes. ...
Context: The lipopolysaccharide (LPS)-induced cytokine release and oxidative stress are validated experimental parameters used to test anti-inflammatory activity. We investigated the effects of homoeopathic mother tinctures, 6 CH, 30 CH and 200 CH dilutions of Arnica montana, Thuja occidentalis and Bryonia alba against LPS (1 μg/ml)-induced cytokine release from RAW-264.7 cells and human whole-blood culture. Materials and Methods: For in vivo evaluations, mice were orally treated with 0.1 ml drug dilutions twice a day for 5 days followed by an intraperitoneal injection of 0.5 mg/kg LPS. After 24 h, the mice were sacrificed and serum levels of pro-inflammatory cytokines and nitric oxide were determined. The extent of oxidative stress was determined in the liver homogenates as contents of reduced glutathione, malondialdehyde, superoxide dismutase and catalase. Results: The tested drug dilutions significantly reduced in vitro LPS-induced release of tumour necrosis factor-α, interleukin-1 (IL-1) and IL-6 from the RAW-264.7 cells and human whole blood culture. Similar suppression of cytokines was evident in mice serum samples. These drugs also protected mice from the LPS-induced oxidative stress in liver tissue. Conclusions: Our findings substantiate the protective effects of Arnica, Thuja and Bryonia homoeopathic dilutions against LPS-induced cytokine elevations and oxidative stress. This study authenticates the claims of anti-inflammatory efficacy of these homoeopathic drugs.
... Studies in the 1990s also demonstrated that environmental stress such as extremely low-dose ionizing radiation and frequency electromagnetic field may influence gene expression. 1,2 In recent decades, evidence from previous studies of several independent groups with different substances, dilutions and experimental designs demonstrated that high and extreme dilutions could alter expression levels of various genes in cells in vitro, [3][4][5][6][7][8] though data from some other studies were not consistent with these findings. 9,10 To our knowledge, most of the studies that focused on gene expression have been conducted with medicines being prepared from a variety of herbal and metal sources. ...
Background Gene expression analysis of cells treated with extreme dilutions or micro amounts of drugs has been used to provide useful suggestions about biological responses. However, most of the previous studies were performed on medicines being prepared from a variety of herbal and metal sources. This study investigated the effects of ultramolecular dilution of the taxane anti-cancer drugs, which are not commonly used in homeopathic medicines, on mRNA expression profiles of five key genes (p53, p21, COX-2, TUBB2A and TUBB3) in the breast cancer cell line MCF-7.
Method MCF-7 cells were exposed to paclitaxel (Taxol) or docetaxel (Taxotere) preparations (6X, 5C and 15C dilutions prepared from pharmacological concentration of 25 nmol/L) for 72 hours. The cell culture groups were evaluated with the trypan blue dye exclusion method for the proliferation/cytotoxicity rates, immuno-staining β-tubulin for microtubule organization, and reverse transcription polymerase chain reaction for gene expression levels.Fold-change in gene expression was determined by the ΔΔCt method.
Results The administration of diluted preparations had little or no cytotoxic effect on MCF-7 cells, but altered the expression of genes analyzed with a complex effect. According to the ΔΔCt method with a five-fold expression difference (p < 0.05) as a cut-off level, ultra-high dilutions of paclitaxel and docetaxel showed differential effects on the studied genes with a concentration-independent activity. Furthermore, the dilutions disrupted the microtubule structure of MCF-7 cells, suggesting that they retain their biological activity.
Conclusion Despite some limitations, our findings demonstrate that gene expression alterations also occur with ultra-high dilutions of taxane drugs.
... Kritische Sichtweisen reichen etwa von dem Einwand, dass Hochpotenzen rein rechnerisch ultramolekulare Verdünnungen seien, weshalb Homöopathie eine Scheintherapie sein müsse, bis zur selektiven Interpretation ausgewählter klinischer Studien im Rahmen der Standards einer «evidence-based medicine» oder der neodogmatischen Ansicht, die Homöopathie erfülle nicht die Voraussetzungen einer «science-based medicine» [10,11]. Der Homöopathie günstige Perspektiven kommen unter anderem aus der Grundlagenforschung, etwa durch neuere Forschungsergebnisse hinsichtlich Nanopartikel [12][13][14][15], Hormesis [16,17], Genexpressivität [18,19], Epigenetik [20] usw. [21,22]. ...
Today, homeopathy, as founded by Samuel Hahnemann (1755- 1843), may be viewed and judged from very different perspectives - resulting from internal differentiation, external critical objections, and research findings from nearly all sciences and humanities. Polyperspectivity is currently overabundant in all areas, also in medicine. In the Western world, its cultural-historical roots are, on the one hand, the traditional monotheistic concept of truth in the Jewish-Christian religion and, on the other hand, the accelerating process of rationalisation and deconstruction within the sciences. The socioeconomic impacts of increasing insecurity and excessive demands range from disorientation and delegitimization of sciences to the hegemony of numbers and algorithms. Options to comply with this challenge are theoretically, for instance, the philosophy of Jainism (Anekantavada, Syadvada, Nayavada) or of Friedrich Nietzsche, or practically, above all, Plato's state philosophy and the Christian doctrine of trinity. Since Hahnemann's conception of homeopathy encompasses the dimensions mythos, logos, and ethos as well, his approach provides a paradigm that has proved for 200 years to handle the dilemma of polyperspectivity in a practical, realistic, and creative way.
... [1,[7][8][9] The authors used Wilcoxon-Mann-Whitney test also in previously reported data, where they used a reverse transcription-PCR analysis on a genome microarray [8] and where a bias in the standard error of mean distribution was also discussed. [10][11][12] According to the authors, any dilution of A. montana in 30% v/v EtOH/water was able to change DEG patterns, with P < 0.05 in Wilcoxon test. [1] Our analysis performed on data from the supplementary tables, [1] using nonparametric Wilcoxon-Mann-Whitney rank test, gave the results showed in Tables 1 and 2. In this calculation, we compared the RPKM of each pooled dilution against the RPKM of the relevant control for each of the 20 or 19 genes for each experiment (Table 1 and Table 2, respectively). ...
Marzotto et al. showed that homeopathic preparations of Arnica montana L. acted directly on gene expression
of Tamm-Horsfall protein-1 (THP-1) monocyte/macrophage cell lines activated with phorbol12-myristate13-
acetate and interleukin-4 (IL-4). A. montana homeopathic dilutions are used in complementary and alternative
medicine to treat inflammation disorders and post-traumatic events as well as for wound repair. The French
Pharmacopoeia of these remedies uses 0.3% ethanol in each centesimal dilution. In this paper, we discuss
how ethanol-containing A. montana homeopathic centesimal dilutions can change gene expression in IL4-
treated monocyte/macrophage THP-1. We assessed the role of ethanol in the Arnica homeopathic dilutions
containing this alcohol by investigating its action on gene expression of THP-1 cell. Evidence would strongly
suggest that the presence of ethanol in these remedies might play a fundamental role in the dilutions ability
to affect gene expression, particularly for doses from 5c to 15c. Where, rather than playing a major role in
the mesoscopic structure of water, the ethanol might have a chemical-physical role in the induction of THP-1
gene expression, apoptosis, and deoxyribonucleic acid function. This evidence generates a debate about the
suggestion that the use of a binary-mixed solvent in homeopathic chemistry, used by Hahnemann since 1810,
may be fundamental to explain the activity of homeopathy on cell models.
... This conclusion might apply to any Arnica dilution exceeding the Avogadro-Loschmidt threshold, yet. 12,13 In conclusion, the paper by Sorrentino et al., should need some reappraisal, in order to assess the experimental reproducibility by checking and revising possible effects from confounders and bias. ...
... To give a possible example of this issue, in a recent paper, statistics was performed using a Friedman sign test, which is less powerful than other non-parametric rank tests, such as the Wil-coxoneMann Withney test [1,18]. This evidence resembles previous reported data, with RT-PCR [6]. According to the authors, any dilution in 30% v/v EtOH/water was able to change DEG patterns, with p < 0.05 in a Wilcoxon test [1]. ...
Basic experimental models in Homeopathy are of major interest because they could get insightful data about the ability of high dilutions to work in a biological system. Due to the extreme difficulty in the highlighting any possible effect and trusting its reliability, methods should be particularly stringent and highly standardized. Confounders, handling process, pre-analytical errors, misleading statistics and misinterpretations may lead to experimental biases. This article tries to elucidate those factors causing bias, taking into account some recent reported evidence in the field.
Background: Homoeopathic medicines have the potential in treatment of patients suffering from cancers, Diabetic Retinopathy, and hemorrhagic disorders wherein anti- angiogenesis plays a vital role. Homoeopathic medicines have been used to treat haemorrhagic disorders. For example, Hamamelis is used in haemorrhages, varicose veins and in venous congestion. Thalaspi bursa is used in flabby condition of uterus, haemorrhagic conditions of uterine fibroid. This been said, their mechanism of action remains unknown. Therefore, this study was intended to establish angiogenic/anti-angiogenic effect of the selected homoeopathic medicines employing chick embryo model. Objective: To study the anti-angiogenic properties of selected homoeopathic medicines employing YSM assay in the chick embryo model. To test the toxicity of selected homoeopathic medicines through Chick embryo model. Materials and Method: Yolk Sac Membrane is widely utilized as an in vivo system to study angiogenesis and anti-angiogenesis. The images for all YSMs were captured and were analyzed for angiogenic/anti-angiogenic effect. Results: Significant anti-angiogenic effects were observed in 10% concentrations of Hamamelis200cH, Thalaspi bursa 200cH and Thalaspi bursa 1M. Reduction of vessel density and total vessel network length, total branching points, total nets and total segment symbolizes that these medicines have anti-angiogenic potential and so they help in treating menstrual disorders, which is followed with cramps and discharge of clots. Conclusion: The mechanism of action of selected homoeopathic medicines as anti- angiogenic agent is proved which can be used to combat various diseases like haemorrhages, varicose veins, in venous congestion and problems related to haemorrhagic disorders. This will further help to study the action of these Homoeopathic medicines as potential line of treatment in patients suffering from cancers, Diabetic Retinopathy, hemorrhagic disorders wherein anti- angiogenesis plays a vital role.
Arnica montana is well known for its anti-inflammatory properties. While the anti-inflammatory activity of Arnica flowers (Arnicae flos) has been extensively studied, that of the whole plant (Arnicae planta tota) is less characterized. We compared the ability of Arnicae planta tota and Arnicae flos extracts to inhibit the pro-inflammatory NF-κB—eicosanoid pathway, using several in vitro and in vivo assays. We showed that Arnicae planta tota inhibited NF-κB reporter activation, with an IC50 of 15.4 μg/mL (vs. 52.5 μg/mL for Arnicae flos). Arnicae planta tota also inhibited LPS-induced expression of ALOX5 and PTGS2 genes in human differentiated macrophages. ALOX5 and PTGS2 encode the 5-lipoxygenase (5-LO) and cyclooxygenase-2 (COX-2) enzymes that initialize the conversion of arachidonic acid into leukotrienes and prostaglandins, respectively. Arnicae planta tota inhibited 5-LO and COX-2 enzymatic activity in vitro and in human primary peripheral blood cells, with lower IC50 compared to Arnicae flos. Finally, Arnicae planta tota applied topically reduced carrageenan-induced mouse paw oedema more efficiently than Arnicae flos. Altogether, Arnicae planta tota displayed a superior anti-inflammatory activity compared to Arnicae flos, suggesting that Arnicae-planta-tota-containing products might be more effective in alleviating the manifestations of acute inflammation than those based on Arnicae flos alone.
La Homeopatía de México | Evidencias Científicas de la Homeopatía | Edición Especial de Aniversario (90 años) | 2023 || EDITORIAL: Para aquellos que demandan evidencias científicas de la Homeopatía. Marcus Zulian Teixeira. | ARTÍCULOS: Homeopatía: una breve descripción de esta especialidad médica. Marcelo Pustiglione, Eduardo Goldenstein, Y. Moisés Checinski. | Formación médica en la terapéutica no convencional en el mundo (Homeopatía y acupuntura). Marcus Zulian Teixeira. | Bases científicas del principio de curación homeopática en la farmacología moderna. Marcus Zulian Teixeira. | La solidez de la investigación homeopática fundamental. Leoni Villano Bonamin. | Efectos de las diluciones homeopáticas altas sobre los modelos in vitro: revisión de la literatura. Silvia Waisse. | Efectos de las diluciones homeopáticas altas sobre las plantas: revisión de la literatura. Marcus Zulian Teixeira, Solange M.T.P.G. Carneiro. | Investigación clínica en Homeopatía: revisiones sistemáticas y estudios clínicos aleatorizados. Silvia Waisse. | Estrógeno potenciado en el tratamiento homeopático del dolor pélvico asociado a endometriosis: Un estudio aleatorizado, doble ciego, controlado con placebo, de 24 semanas. Marcus Zulian Teixeira, Sérgio Podgaec, Edmund Chada Baracat. | Estudio aleatorizado, doble ciego, sobre la eficacia del tratamiento homeopático en niños con amigdalitis recurrente. Sérgio E. Furuta, Luc L. M. Weckx, Cláudia R. Figueiredo. | ¿Los medicamentos homeopáticos provocan agravación o efectos adversos dependientes de los fármacos? Flávio Dantas. | ¿Los medicamentos homeopáticos inducen síntomas en voluntarios aparentemente sanos? Contribución brasileña al debate sobre los estudios patogénicos homeopáticos. Flávio Dantas. |
La Homeopatía de México | Evidencias Científicas de la Homeopatía | Edición Especial de Aniversario (90 años) | 2023 ||
EDITORIAL:
Para aquellos que demandan evidencias científicas de la Homeopatía.
Marcus Zulian Teixeira. |
ARTÍCULOS:
Homeopatía: una breve descripción de esta especialidad médica.
Marcelo Pustiglione, Eduardo Goldenstein, Y. Moisés Checinski. |
Formación médica en la terapéutica no convencional en el mundo (Homeopatía y acupuntura).
Marcus Zulian Teixeira. |
Bases científicas del principio de curación homeopática en la farmacología moderna.
Marcus Zulian Teixeira. |
La solidez de la investigación homeopática fundamental.
Leoni Villano Bonamin. |
Efectos de las diluciones homeopáticas altas sobre los modelos in vitro: revisión de la literatura.
Silvia Waisse. |
Efectos de las diluciones homeopáticas altas sobre las plantas: revisión de la literatura.
Marcus Zulian Teixeira, Solange M.T.P.G. Carneiro. |
Investigación clínica en Homeopatía: revisiones sistemáticas y estudios clínicos aleatorizados.
Silvia Waisse. |
Estrógeno potenciado en el tratamiento homeopático del dolor pélvico asociado a endometriosis: Un estudio aleatorizado, doble ciego, controlado con placebo, de 24 semanas.
Marcus Zulian Teixeira, Sérgio Podgaec, Edmund Chada Baracat. |
Estudio aleatorizado, doble ciego, sobre la eficacia del tratamiento homeopático en niños con amigdalitis recurrente.
Sérgio E. Furuta, Luc L. M. Weckx, Cláudia R. Figueiredo. |
¿Los medicamentos homeopáticos provocan agravación o efectos adversos dependientes de los fármacos?
Flávio Dantas. |
¿Los medicamentos homeopáticos inducen síntomas en voluntarios aparentemente sanos? Contribución brasileña al debate sobre los estudios patogénicos homeopáticos.
Flávio Dantas. |
This study was aimed to evaluate therapeutic effects of thymoquinone on male reproductive damages induced by paclitaxel. Forty‐eight male rats were divided; control, paclitaxel (4 mg/kg), paclitaxel + thymoquinone (1.25, 2.5 and 5 mg/kg) and thymoquinone (1.25, 2.5 and 5 mg/kg). Paclitaxel and thymoquinone were administrated intraperitoneally for 4 and 14 days respectively. Then, the testes were removed for H&E staining, sperm parameters and apoptotic genes expression assessments. Serum levels of nitric oxide, total antioxidant capacity and testosterone were evaluated, and sperm DNA fragmentation was assessed. Paclitaxel significantly (p < .05) increased nitric oxide, decreased total antioxidant capacity and reduced testosterone levels than control group. Sperm motility, viability and count were significantly (p < .05) reduced in paclitaxel group than control. Co‐administration of thymoquinone + paclitaxel caused decreased levels of nitric oxide and increased total antioxidant capacity, testosterone levels and reproductive parameters than paclitaxel group significantly (p < .05). Paclitaxel significantly (p < .05) increased caspase‐3 and p‐53 and decreased Bcl‐2 genes expression than control. Sperm DNA fragmentation index was also increased significantly (p < .05) in paclitaxel group than control, and this value was decreased in whole doses of paclitaxel + thymoquinone groups than paclitaxel. Thymoquinone can alleviate the side effects of paclitaxel on the male reproductive system.
Introduction: In addition to the four pillars of homeopathy, vitalism and the miasmatic theory are often used to explain the health–disease process. According to Hahnemann's concepts, homeopathic miasms are the main obstacle to the cure of chronic diseases, with psora being the fundamental cause of all forms of diseases. According to modern genetics, the disease-promoting epigenetic alterations are the fundamental cause of the manifestation of chronic diseases.
Objective: This article develops a philosophical–scientific correlation between chronic miasms and disease-promoting epigenetic modifications, aiming to justify the isopathic use of auto-sarcode of an individual's DNA as homeopathic medicine.
Results: Based on the study of homeopathic doctrine and epigenetics, a conceptual and functional correlation is observed between homeopathic chronic miasms and disease-promoting epigenetic modifications. Additionally, several experimental studies suggest that homeopathy's mechanism of action may be by modulating gene expression.
Conclusions: By the philosophical–scientific correlations described, it is inferred that disease-promoting epigenetic alterations are the biological representation of the chronic miasms, suggesting the isopathic use of auto-sarcode of DNA as homeopathic therapeutic modulator of gene expression for the management of chronic diseases.
Para desmistificar a falácia ou pós-verdade de que “não existem evidências científicas em homeopatia”, disseminada indistintamente em todos os meios, a Câmara Técnica de Homeopatia do Conselho Regional de Medicina do Estado de São Paulo (CREMESP) elaborou o Dossiê Especial “Evidências Científicas em Homeopatia”, que acaba de ser disponibilizado online na Revista de Homeopatia da Associação Paulista de Homeopatia (APH).
Englobando 9 revisões (e 2 RCT) sobre diversas linhas de pesquisa existentes nos mais variados campos da ciência, as quais comportam centenas de artigos científicos publicados em periódicos distintos, esse dossiê evidencia à classe médica e científica, assim como ao público em geral, o estado da arte da pesquisa homeopática.
Apesar das dificuldades e limitações existentes para o desenvolvimento de pesquisas na área, tanto pelos aspectos metodológicos quanto pela ausência de apoio institucional e financeiro, o conjunto de estudos experimentais e clínicos citados, que fundamentam os pressupostos homeopáticos e confirmam a eficácia e a segurança da terapêutica, é prova inconteste de que “existem evidências científicas em homeopatia”, ao contrário do preconceito falsamente disseminado.
Com a elaboração e a divulgação desse dossiê, sob os auspícios da Câmara Técnica de Homeopatia do CREMESP, esperamos esclarecer e sensibilizar os colegas de profissão sobre a validade e a importância do emprego da homeopatia como prática médica adjuvante e complementar às demais especialidades, segundo princípios éticos e seguros, a fim de se ampliar o entendimento do processo de adoecimento humano e o arsenal terapêutico, incrementar o ato médico e sua resolutividade nas doenças crônicas, minimizar os efeitos adversos dos fármacos modernos e fortalecer a relação médico-paciente, dentre outros aspectos. Dessa forma, poderemos trabalhar unidos em torno da “mais elevada e única missão do médico que é tornar saudáveis as pessoas doentes, o que se chama curar” (Samuel Hahnemann, Organon da arte de curar, § 1).
In July 2017, to demystify the fallacy – or post-truth – asserting “there are no scientific evidence for homeopathy”, the Technical Chamber for Homeopathy, Regional Medical Council of the State of São Paulo (CREMESP, Brazil) published the Special Dossier “Evidências Científicas em Homeopatia”, available online in the scientific journal of the São Paulo Homeopathic Medical Association (APH), Revista de Homeopatia.
After this publication, in view of the request of homeopathic doctors and institutions from other countries, the Technical Chamber for Homeopathy, CREMESP, produced an edition of the dossier in English, “Scientific Evidence for Homeopathy”, which has just been made available online in the Revista de Homeopatia.
Encompassing 9 revisions (and 2 randomized clinical trials) on several lines of homeopathic research, containing hundreds of scientific articles published in various journals, this dossier highlights to the scientific and medical class, as well as to the general public, the state of the art of homeopathic research:
Despite the ongoing difficulties and limitations opposing the development of research in homeopathy – partly due to methodological aspects, and partly to lack of institutional and financial support – the experimental and clinical studies described in this dossier, which ground the homeopathic assumptions and confirm the efficacy and safety of this approach to therapeutics – provide unquestionable proof for the “availability of scientific evidence for homeopathy”, against the false and prejudiced opinion that is widely divulgated.
With the divulgation of the present dossier, prepared with the support of Technical Chamber for Homeopathy, CREMESP, we hope to dispel doubts and sensitize our colleagues as to the validity and relevance of homeopathy as adjuvant treatment complementary to all other medical specialties according to ethical and safe principles. Our overall goals are to broaden the understanding of human disease, increase the therapeutic resources, contribute to the definition and effectiveness of medicine in chronic diseases, minimize the adverse effects of modern drugs and strengthen the patient-doctor relationship, among other aspects. In this way, we will be able to work together, since “The physician’s high and only mission is to restore the sick to health, to cure, as it is termed (Samuel Hahnemann, Organon of medicine, § 1).
Palliative care approaches that effectively manage distressful symptoms associated with wounds at the end of life remain elusive. This 4-week study examined a topical wound powder RGN107 for reducing pain, odor, and exudate in 50 patients with pressure ulcers, skin tears, and malignant/fungating and vascular wounds receiving hospice or palliative care and explored quality of life for the caregiver. Through an observational design, the outcomes were measured with visual analog scales, 2 pain questionnaires, and a caregiver quality-of-life instrument. Intent-to-treat analyses were used. Statistically significant reductions in pain (P =.001), odor (P =.04), and exudate (P =.00003) were observed. Caregiver quality of life remained unchanged (P =.28); however, improvements were noted in 3 subscales. Findings suggest topical RGN107 reduced pain, odor, and exudate in a highly challenged population with wounds at the end of life. A larger comparative effectiveness trial should be conducted with other wound powder comparators and usual care approaches and should include cost benefits.
Homeopathic arnica is widely believed to control bruising, reduce swelling and promote recovery after local trauma; many patients therefore take it perioperatively. To determine whether this treatment has any effect, we conducted a double-blind, placebo-controlled, randomized trial with three parallel arms. 64 adults undergoing elective surgery for carpal tunnel syndrome were randomized to take three tablets daily of homeopathic arnica 30C or 6C or placebo for seven days before surgery and fourteen days after surgery. Primary outcome measures were pain (short form McGill Pain Questionnaire) and bruising (colour separation analysis) at four days after surgery. Secondary outcome measures were swelling (wrist circumference) and use of analgesic medication (patient diary).
62 patients could be included in the intention-to-treat analysis. There were no group differences on the primary outcome measures of pain ( P=0.79) and bruising ( P=0.45) at day four. Swelling and use of analgesic medication also did not differ between arnica and placebo groups. Adverse events were reported by 2 patients in the arnica 6C group, 3 in the placebo group and 4 in the arnica 30C group.
The results of this trial do not suggest that homeopathic arnica has an advantage over placebo in reducing postoperative pain, bruising and swelling in patients undergoing elective hand surgery.
Alendronate (ALN) is an antiresorptive agent widely used for the treatment of osteoporosis. Its suppressive effect on osteoclasts has been extensively studied. However, the effect of ALN on bone formation is not as clear as its effect on resorption. The objective was to determine the effect of short-term ALN on bone formation and tooth extraction wound healing. Molar tooth extractions were performed in mice. ALN, parathyroid hormone (PTH), or saline (vehicle control) was administered. PTH was used as the bone anabolic control. Mice were euthanized at 3, 5, 7, 10, and 21 d after extractions. Hard tissue healing was determined histomorphometrically. Neutrophils and lymphatic and blood vessels were quantified to evaluate soft tissue healing. Gene expression in the wounds was assessed at the RNA level. Furthermore, the vossicle bone transplant system was used to verify findings from extraction wound analysis. Alkaline phosphatase (ALP) was visualized in the vossicles to assess osteoblast activity. ALN exhibited no negative effect on bone formation. In intact tibiae, ALN increased bone mass significantly more than PTH did. Consistently, significantly elevated osteoblast numbers were noted. In the extraction sockets, bone fill in the ALN-treated mice was equivalent to the control. Genes associated with bone morphogenetic protein signaling, such as bmp2, nog, and dlx5, were activated in the extraction wounds of the ALN-treated animals. Bone formation in vossicles was significantly enhanced in the ALN versus PTH group. In agreement with this, ALN upregulated ALP activity considerably in vossicles. Neutrophil aggregation and suppressed lymphangiogenesis were evident in the soft tissue at 21 d after extraction, although gross healing of extraction wounds was uneventful. Bone formation was not impeded by short-term ALN treatment. Rather, short-term ALN treatment enhanced bone formation. ALN did not alter bone fill in extraction sockets.
Arnica montana plant has been used in folklore medicine to alleviate pain, inflammation, swelling of muscles and joints associated with rheumatoid arthritis and other inflammatory conditions. The present study aims to investigate the therapeutic effects and mechanism of action of Arnica montana flower methanol extract (AMME) against both inflammation and oxidative stress in collagen-induced arthritis rat model.
Oral administration of AMME was found to reduce clinical signs and improved histological and radiological status of the hind-limb joints. AMME-treated rats had lower expression levels of nitric oxide, tumor necrosis factor-α, interleukins (IL-1β, IL-6 and IL-12), and titer of type II anti-collagen antibody as compared to untreated CIA rats. Furthermore, by inhibiting the above mediators, AMME also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage.
The alleviation of arthritis in rats can be very likely due to the combined action of phenolic and flavonoid compounds, which are found to be the major constituents in our GC-MS analysis. This study also shed some light on mechanisms involved in diminution of inflammatory mediators, free radical generating toxicants, and enhancement of antioxidant armory, thereby preventing further tissue damage, injury and synovial hyper proliferation in arthritis.
This article is protected by copyright. All rights reserved.
It has been claimed that the homeopathic principle of 'similarity' (or 'similia') and the use of individualized remedies in extremely low doses conflicts with scientific laws, but this opinion can be disputed on the basis of recent scientific advances. Several mechanisms to explain the responsiveness of cells to ultra-low doses and the similarity as inversion of drug effects, have again been suggested in the framework of hormesis and modern paradoxical pharmacology. Low doses or high dilutions of a drug interact only with the enhanced sensitivities of regulatory systems, functioning as minute harmful stimuli to trigger specific compensatory healing reactions. Here we review hypotheses about homeopathic drug action at cellular and molecular levels, and present a new conceptual model of the principle of similarity based on allosteric drug action. While many common drugs act through orthostatic chemical interactions aimed at blocking undesired activities of enzymes or receptors, allosteric interactions are associated with dynamic conformational changes and functional transitions in target proteins, which enhance or inhibit specific cellular actions in normal or disease states. The concept of allostery and the way it controls physiological activities can be broadened to include diluted/dynamized compounds, and may constitute a working hypothesis for the study of molecular mechanisms underlying the inversion of drug effects.
Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.
The mechanisms of monocyte recruitment to arteriogenic collaterals are largely unknown. We investigated the role of chemokine (C-X-C-motif) ligand 1 (CXCL1) and its cognate receptor, chemokine (C-X-C-motif) receptor 2 (CXCR2) in arteriogenesis.
After femoral artery ligation in Sprague-Dawley rats, either native collaterals were harvested or placebo, CXCL1 or CXCR2 blocker was administered via an osmopump. Perfusion recovery was measured with Laser Doppler, leukocyte populations were analyzed by fluorescence-activated cell sorting, and hind limb sections were stained for macrophage marker cluster of differentiation 68 (CD68). In vitro, fluorescent CXCL1 or human acute monocytic leukemia cell line (THP-1) monocytic cells were flown over shear-stressed endothelium. CXCL1 mRNA expression in collaterals was dramatically upregulated already 1 h after ligation (ratio ligated/sham 5.73). CD68 mRNA was upregulated from 12 h until 3 days after ligation (peak ratio ligated/sham 2.65). CXCL1 treatment augmented perfusion recovery at 3 and 7 days (p < 0.05) after ligation, and a significant increase in the number of peri-collateral macrophages was evident concomitantly (p < 0.05). Conversely, CXCR2 antagonist treatment caused a decrease in perfusion recovery both at 7 and 10 days postligation (p = 0.01) and also significantly reduced the number of peri-collateral macrophages (p < 0.05). In vitro, CXCL1 tethered to and was taken up by endothelial cells under shear stress conditions and enhanced THP-1 adherence compared to control (p < 0.05). In contrast, CXCR2 antagonist compromised THP-1 adherence to endothelial cells (p < 0.05).
CXCL1 presented on the luminal endothelial surface leads to an increase in the number of peri-collateral macrophages, thus improving the arteriogenic response after arterial ligation.
Plant systems are useful research tools to address basic questions in homeopathy as they make it possible to overcome some of the drawbacks encountered in clinical trials (placebo effect, ethical issues, duration of the experiment, and high costs). The objective of the present study was to test the hypothesis whether 7-day-old wheat seedlings, grown from seeds either poisoned with a sublethal dose of As2O3 or unpoisoned, showed different significant gene expression profiles after the application of ultrahigh diluted As2O3 (beyond Avogadro's limit) compared to water (control). The results provided evidence for a strong gene modulating effect of ultrahigh diluted As2O3 in seedlings grown from poisoned seeds: a massive reduction of gene expression levels to values comparable to those of the control group was observed for several functional classes of genes. A plausible hypothesis is that ultrahigh diluted As2O3 treatment induced a reequilibration of those genes that were upregulated during the oxidative stress by bringing the expression levels closer to the basal levels normally occurring in the control plants.
Arnica montana has been widely used as a homeopathic remedy for the treatment of several inflammatory conditions in pain management and postoperative settings. This review gives an overview of the therapeutic use of Arnica montana in the above-mentioned fields also focusing on its mechanisms of action learned from animal models and in vitro studies. Arnica montana is more effective than placebo when used for the treatment of several conditions including post-traumatic and postoperative pain, edema, and ecchymosis. However, its dosages and preparations used have produced substantial differences in the clinical outcome. Cumulative evidence suggests that Arnica montana may represent a valid alternative to non-steroidal anti-inflammatory drugs, at least when treating some specific conditions.
Immunoglobulin E (IgE) is pathogenic in allergic diseases such as asthma, allergic rhinitis, atopic dermatitis, and food allergy. Recent studies using genetically modified IgE reporter mice indicate that the majority of serum IgE in mice is produced by short-lived IgE plasma cells, with minor contributions from long-lived IgE plasma cells, and implicate IgG1 and IgE memory B cells as potential sources of IgE memory. Clinical studies using antibodies against IL-13 or the IL-4 and IL-13 receptor subunit IL-4Rα, as well as an antibody against the M1 prime domain of human membrane IgE, indicate that, similar to mice, a proportion of IgE in humans is derived from ongoing IgE immune responses and short-lived plasma cells. Targeting IgE production may lead to new therapies for the treatment of allergic diseases.
Background
Fe toxicity occurs in lowland rice production due to excess ferrous iron (Fe2+) formation in reduced soils. To contribute to the breeding for tolerance to Fe toxicity in rice, we determined quantitative trait loci (QTL) by screening two different bi-parental mapping populations under iron pulse stresses (1,000 mg L−1 = 17.9 mM Fe2+ for 5 days) in hydroponic solution, followed by experiments with selected lines to determine whether QTLs were associated with iron exclusion (i.e. root based mechanisms), or iron inclusion (i.e. shoot-based mechanisms).
Results
In an IR29/Pokkali F8 recombinant inbred population, 7 QTLs were detected for leaf bronzing score on chromosome 1, 2, 4, 7 and 12, respectively, individually explaining 9.2-18.7% of the phenotypic variation. Two tolerant recombinant inbred lines carrying putative QTLs were selected for further experiments. Based on Fe uptake into the shoot, the dominant tolerance mechanism of the tolerant line FL510 was determined to be exclusion with its root architecture being conducive to air transport and thus the ability to oxidize Fe2+ in rhizosphere. In line FL483, the iron tolerance was related mainly to shoot-based mechanisms (tolerant inclusion mechanism). In a Nipponbare/Kasalath/Nipponbare backcross inbred population, 3 QTLs were mapped on chromosomes 1, 3 and 8, respectively. These QTLs explained 11.6-18.6% of the total phenotypic variation. The effect of QTLs on chromosome 1 and 3 were confirmed by using chromosome segment substitution lines (SL), carrying Kasalath introgressions in the genetic background on Nipponbare. The Fe uptake in shoots of substitution lines suggests that the effect of the QTL on chromosome 1 was associated with shoot tolerance while the QTL on chromosome 3 was associated with iron exclusion.
Conclusion
Tolerance of certain genotypes were classified into shoot- and root- based mechanisms. Comparing our findings with previously reported QTLs for iron toxicity tolerance, we identified co-localization for some QTLs in both pluse and chronic stresses, especially on chromosome 1.
A dark brown polymeric complex was isolated from flowering parts of medicinal plant Arnica montana L. by hot alkaline extraction followed by neutralization and multi-step extractions with organic solvents. It was recovered in 5.7% yield, on GPC showed two peaks of molecular mass of 9 and 3.5 kDa. The compositional analyses of Arnica complex revealed the presence of carbohydrates (26%), uronic acids (12%), phenolics (1.25mM or 213mg of GAE/1g), and low protein content (∼1%). The carbohydrate moiety was rich mainly in rhamnogalacturonan and arabinogalactan. The antitussive tests showed the reduction of the cough efforts by Arnica complex, however, its total antitussive effect was lower compared with that of codeine, the strongest antitussive agent. The bronchodilatory activity of Arnica complex was similar to salbutamol, a classic antiasthmatic drug, and was confirmed by significantly decreased values of specific airways resistance in vivo and by considerably attenuated the amplitude of acetylcholine and histamine-induced contractions in vitro. Arnica complex did not show any cytotoxic effect on mouse fibroblast cultures and human lung cells, up to the dose of 500μg/mL.
The capacity to regenerate complex tissue structures after amputation in humans is limited to the digit tip. In a comparable mouse digit model, which includes both distal regeneration-competent and proximal regeneration-incompetent regions, successful regeneration involves precise orchestration of complex microenvironmental cues including paracrine signaling via heterogeneous cell-cell interactions. Initial cellular processes, such as proliferation and migration, are critical in the formation of an initial stable cell mass and the ultimate regenerative outcome. Hence, the objective of these in vitro studies was to investigate the effect of soluble factors secreted by fibroblasts and mesenchymal stem cells (MSCs) on the proliferation and migration of cells from the regeneration-competent (P3) and -incompetent (P2) regions of the mouse digit tip. We found that P2 and P3 cells were more responsive to fibroblasts than MSCs and that the effects were mediated by bi-directional communication. To initiate understanding of the specific soluble factors that may be involved in the fibroblast-mediated changes in migration of P2 and P3 cells, bone morphogenic protein 2 (BMP2) was exogenously added to the medium. We found that changes in migration of P3 cells were similar when exposed to BMP2 or co-cultured with fibroblasts, indicating that BMP signaling may be responsible for the migratory response of P3 cells to the presence of fibroblasts. Furthermore, BMP2 expression in fibroblasts was shown to be responsive to tensile strain, as is present during wound closure. Therefore, these in vitro studies indicate that regenerative processes may be regulated by fibroblast-secreted soluble factors, which in turn are modulated by both crosstalk between heterogeneous phenotypes and the physical microenvironment of the healing site.
Diabetes mellitus results from immune-cell invasion into pancreatic islets of Langerhans, eventually leading to selective destruction of the insulin-producing β-cells. How this process is initiated is not well understood. In this study, we investigated the regulation of the CXCL1 and CXCL2 genes, which encode proteins that promote migration of CXCR(2+) cells, such as neutrophils, towards secreting tissue. Herein, we found that IL-1β markedly enhanced the expression of the CXCL1 and CXCL2 genes in rat islets and β-cell lines which resulted in increased secretion of each of these proteins. CXCL1 and CXCL2 also stimulated the expression of specific integrin proteins on the surface of human neutrophils. Mutation of a consensus NF-κB genomic sequence present in both gene promoters reduced the ability of IL-1β to promote transcription. In addition, IL-1β induced binding of the p65 and p50 subunits of NF-κB to these consensus κB regulatory elements as well as to additional κB sites located near the core promoter regions of each gene. Additionally, serine phosphorylated STAT1 bound to the promoters of the CXCL1 and CXCL2 genes. We further found that IL-1β induced specific post-translational modifications to histone H3 in a time frame congruent with transcription factor binding and transcript accumulation. We conclude that IL-1β-mediated regulation of the CXCL1 and CXCL2 genes in pancreatic β-cells requires stimulus-induced changes in histone chemical modifications, recruitment of the NF-κB and STAT1 transcription factors to genomic regulatory sequences within the proximal gene promoters and increases in phosphorylated forms of RNA polymerase II.
Homeopathy is controversial, due to the claims made for very high dilutions. Although several theories are proposed to understand the mechanisms of action, none are scientifically verified. This study aimed to investigate the efficacy of the selected homeopathic medicines in specific in vitro cancer models.
We assessed the cytotoxic activity of selected homeopathic medicines in mother tincture (MT), and ultramolecular dilution (30C, 200C, 1M and 10M) against cell lines deriving from tumors of particular organs, Sarsaparilla (Sars) on ACHN cells (human renal adenocarcinoma), Ruta graveolens (Ruta) on COLO-205 (human colorectal carcinoma), and Phytolacca decandra (Phyto) on MCF-7 (human breast carcinoma). Sars was also tested against Madin-Darby canine kidney (MDCK) cells (a non-malignant cell line). Cytotoxicity was measured using the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) method, anti-proliferative activity by trypan blue exclusion assay, apoptosis determined by dual staining the cells with ethidium bromide (EB) and acridine orange (AO) dyes.
MTs and ultra-diluted preparations of the three homeopathic medicines had highly significant effects in the respective cancer cell lines, producing cytotoxicity and a decrease in cell proliferation. The effects were greatest with the MTs, but in all cases and persisted, although to a lesser degree in the ultra-diluted molecular preparations. Sars showed no effect on MDCK cells. In the homeopathic medicine treated cultures, hallmarks of apoptosis were evident including, cell shrinkage, chromatin condensation and DNA fragmentation.
This study provides preliminary laboratory evidence indicating the ability of homeopathic medicines as anticancer agents. Further studies of the action of these homeopathic remedies are warranted.
Arnica montana L. and Artemisia absinthium L. (Asteraceae) are medicinal plants native to temperate regions of Europe, including Romania, traditionally used for treatment of skin wounds, bruises and contusions. In the present study, A. montana and A. absinthium ethanolic extracts were evaluated for their chemical composition, antioxidant activity and protective effect against H2O2-induced oxidative stress in a mouse fibroblast-like NCTC cell line.
A. absinthium extract showed a higher antioxidant capacity than A. montana extract as Trolox equivalent antioxidant capacity, Oxygen radical absorbance capacity and 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging activity, in correlation with its flavonoids and phenolic acids content. Both plant extracts had significant effects on the growth of NCTC cells in the range of 10-100 mg/L A. montana and 10-500 mg/L A. absinthium. They also protected fibroblast cells against hydrogen peroxide-induced oxidative damage, at the same doses. The best protection was observed in cell pre-treatment with 10 mg/L A. montana and 10-300 mg/L A. absinthium, respectively, as determined by Neutral red and lactate dehydrogenase assays. In addition, cell pre-treatment with plant extracts, at these concentrations, prevented morphological changes induced by hydrogen peroxide. Flow-cytometry analysis showed that pre-treatment with A. montana and A. absinthium extracts restored the proportion of cells in each phase of the cell cycle.
A. montana and A. absinthium extracts, rich in flavonoids and phenolic acids, showed a good antioxidant activity and cytoprotective effect against oxidative damage in fibroblast-like cells. These results provide scientific support for the traditional use of A. montana and A. absinthium in treatment of skin disorders.
Concussive injury (or mild traumatic brain injury; mTBI) can exhibit features of focal or diffuse injury patterns. We compared/contrasted the cellular and molecular responses after mild controlled cortical impact (CCI; a focal injury) or fluid percussion injury (FPI; a diffuse injury) in rats. The rationale for this comparative analysis was to investigate the brain's response to mild diffuse versus mild focal injury, to identify common molecular changes triggered by these injury modalities, and to determine the functional pathways altered after injury that may provide novel targets for therapeutic intervention. Microarrays containing probes against 21,792 unique mRNAs were used to investigate the changes in cortical mRNA expression levels at 3 and 24 hr post-injury. Of the 354 mRNAs with signifincaly altered expression levels after mCCI, over 89% (316 mRNAs) were also contained within the mFPI data set. However, mFPI initiated a more widespread molecular response, with over 2300 mRNAs differentially expressed. Bioinformatic analysis of annotated Gene Ontology molecular function and biological pathway terms showed a significant overrepresentation of genes belonging to inflammation, stress and signaling categories in both datasets. We therefore examined changes in the protein levels of a panel of 23 cytokines and chemokines using a Luminex-based bead immunoassay, and detected significant increases in MIP-1α (CCL3), GRO-KC (CXCL1), IL-1α, IL-1β, and IL-6. Immunohistochemical localization of MIP-1α and IL-1β showed marked increases at 3 hr post-injury in the cortical vasculature and microglia, respectively, that were largely resolved by 24 hr post-injury. Our findings demonstrate that both focal and diffuse mild TBI triggered many shared pathobiological processes (e.g. inflammatory responses) that could be targeted for mechanism-based therapeutic interventions.
Background
Drugs of plant origin such as Arnica montana, Calendula officinalis or Hypericum perforatum have been frequently used to promote wound healing. While their effect on wound healing using preparations at pharmacological concentrations was supported by several in vitro and clinical studies, investigations of herbal homeopathic remedies on wound healing process are rare. The objective of this study was to investigate the effect of a commercial low potency homeopathic remedy Similasan® Arnica plus Spray on wound closure in a controlled, blind trial in vitro.
Methods
We investigated the effect of an ethanolic preparation composed of equal parts of Arnica montana 4x, Calendula officinalis 4x, Hypericum perforatum 4x and Symphytum officinale 6x (0712–2), its succussed hydroalcoholic solvent (0712–1) and unsuccussed solvent (0712–3) on NIH 3T3 fibroblasts. Cell viability was determined by WST-1 assay, cell growth using BrdU uptake, cell migration by chemotaxis assay and wound closure by CytoSelect ™Wound Healing Assay Kit which generated a defined “wound field”. All assays were performed in three independent controlled experiments.
Results
None of the three substances affected cell viability and none showed a stimulating effect on cell proliferation. Preparation (0712–2) exerted a stimulating effect on fibroblast migration (31.9%) vs 14.7% with succussed solvent (0712–1) at 1:100 dilutions (p < 0.001). Unsuccussed solvent (0712–3) had no influence on cell migration (6.3%; p > 0.05). Preparation (0712–2) at a dilution of 1:100 promoted in vitro wound closure by 59.5% and differed significantly (p < 0.001) from succussed solvent (0712–1), which caused 22.1% wound closure.
Conclusion
Results of this study showed that the low potency homeopathic remedy (0712–2) exerted in vitro wound closure potential in NIH 3T3 fibroblasts. This effect resulted from stimulation of fibroblasts motility rather than of their mitosis.
Topical application of dexpanthenol is widely used in clinical practice for the improvement of wound healing. Previous in vitro experiments identified a stimulatory effect of pantothenate on migration, proliferation and gene regulation in cultured human dermal fibroblasts. To correlate these in vitro findings with the more complex in vivo situation of wound healing, a clinical trial was performed in which the dexpanthenol-induced gene expression profile in punch biopsies of previously injured and dexpanthenol-treated skin in comparison to placebo-treated skin was analyzed at the molecular level by Affymetrix® GeneChip analysis. Upregulation of IL-6, IL-1β, CYP1B1, CXCL1, CCL18 and KAP 4-2 gene expression and downregulation of psorasin mRNA and protein expression were identified in samples treated topically with dexpanthenol. This in vivo study might provide new insight into the molecular mechanisms responsible for the effect of dexpanthenol in wound healing and shows strong correlations to previous in vitro data using cultured dermal fibroblasts.
Dendritic cell-specific intercellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN) is an important pattern recognition receptor on dendritic cells (DCs), and its expression shows significant cytological and histological specificity, being interleukine-4 (IL-4) dependent. The signaling pathways through which IL-4 regulates expression of DC-SIGN are still unclear. We used phorbol 12-myristate 13-acetate- (PMA-) differentiated THP-1 cells as the in vitro model of monocyte/macrophage cells to study the signaling pathways involved in IL-4-regulated expression of DC-SIGN. We found that a high expression of DC-SIGN could be induced by IL-4 at the levels of mRNA and cell surface protein. Upregulated expression of DC-SIGN was almost completely blocked by the specific inhibitor of ERK pathway, and partly reduced by the specific inhibitors of JAK-STAT and NF-κB pathways. The activation of the three signaling pathways was directly confirmed by testing the phosphorylation of protein kinase within the cytoplasm and nucleus over time. The analysis of cis-acting elements of DC-SIGN promoter showed that the activity of DC-SIGN promoter without Ets-1 transcription factors binding site almost completely disappeared. Our results demonstrated that multiple signaling pathways are involved in IL-4 induced high expression of DC-SIGN on THP-1 cells, in which ERK pathway is the main signaling pathway and mediated by the Ets-1 transcription factors binding site.
Activation of microglia causes the production of proinflammatory factors and upregulation of NADPH oxidase (NOX) that form reactive oxygen species (ROS) that lead to neurodegeneration. Previously, we reported that 10 daily doses of ethanol treatment induced innate immune genes in brain. In the present study, we investigate the effects of chronic ethanol on activation of NOX and release of ROS, and their contribution to ethanol neurotoxicity.
Male C57BL/6 and NF-κB enhanced GFP mice were treated intragastrically with water or ethanol (5 g/kg, i.g., 25% ethanol w/v) daily for 10 days. The effects of chronic ethanol on cell death markers (activated caspase-3 and Fluoro-Jade B), microglial morphology, NOX, ROS and NF-κB were examined using real-time PCR, immunohistochemistry and hydroethidine histochemistry. Also, Fluoro-Jade B staining and NOX gp91phox immunohistochemistry were performed in the orbitofrontal cortex (OFC) of human postmortem alcoholic brain and human moderate drinking control brain.
Ethanol treatment of C57BL/6 mice showed increased markers of neuronal death: activated caspase-3 and Fluoro-Jade B positive staining with Neu-N (a neuronal marker) labeling in cortex and dentate gyrus. The OFC of human post-mortem alcoholic brain also showed significantly more Fluoro-Jade B positive cells colocalized with Neu-N, a neuronal marker, compared to the OFC of human moderate drinking control brain, suggesting increased neuronal death in the OFC of human alcoholic brain. Iba1 and GFAP immunohistochemistry showed activated morphology of microglia and astrocytes in ethanol-treated mouse brain. Ethanol treatment increased NF-κB transcription and increased NOX gp91phox at 24 hr after the last ethanol treatment that remained elevated at 1 week. The OFC of human postmortem alcoholic brain also had significant increases in the number of gp91phox + immunoreactive (IR) cells that are colocalized with neuronal, microglial and astrocyte markers. In mouse brain ethanol increased gp91phox expression coincided with increased production of O2- and O2- - derived oxidants. Diphenyleneiodonium (DPI), a NOX inhibitor, reduced markers of neurodegeneration, ROS and microglial activation.
Ethanol activation of microglia and astrocytes, induction of NOX and production of ROS contribute to chronic ethanol-induced neurotoxicity. NOX-ROS and NF-κB signaling pathways play important roles in chronic ethanol-induced neuroinflammation and neurodegeneration.
To examine to what degree an ultra-highly diluted homeopathic remedy, Arnica Montana 30C (AM-30C), used in the treatment of shock and injury, can modulate the expression of nucleotide excision repair genes in Escherichia coli exposed to ultraviolet (UV) irradiation.
E. coli were cultured to their log phase in a standard Luria-Bertani medium and then exposed to sublethal doses of UV irradiation at 25 and 50 J/m(2) for 22.5 and 45 s, respectively. The UV-exposed bacteria were then supplemented with either AM-30C (drug) or placebo (P-30C). The drug-treated and placebo-treated bacteria were subjected to assay for DNA damage and oxidative stress 90 min after UV exposure. Several protocols like comet assay, gel electrophoresis for DNA ladder and intracellular reactive oxygen species (ROS) generation, and biomarker measurement like superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were conducted. The mRNA expressions of the excision repair genes like ultraviolet repair uvrA, B and C genes (or also known as excision repair genes) were estimated by reverse transcription-polymerase chain reaction method.
The UV-exposed bacteria showed DNA damage and oxidative stress, as revealed by an increase in ROS generation, and a decrease in SOD, CAT and GSH activities. As compared to placebo, the AM-30C-treated bacteria showed less DNA damage and oxidative stress as manifested by a decrease in ROS generation, and an increase in SOD, CAT and GSH activities. AM-30C also up-regulated the expression of repair genes as compared to the control.
AM-30C helped repair the DNA damage through up-regulation of repair genes and also ameliorated the oxidative stress through the reduction of ROS generation and suitable modulation of anti-oxidative stress enzymes.
Homoeopathic medicines treat diseases, including cancer, using ultradiluted preparations. Earlier studies indicated that homoeopathic medicines are cytotoxic to tumor cells and reduced animal tumors. However, the mechanism of homoeopathic medicines at the cellular level is not known.
The following drugs were used in the study: Ruta 200C, Carcinosinum 200C, Hydrastis 200C, Thuja 200C, and Thuja 1M. These drugs were tested for their ability to induce apoptosis as seen by morphology, DNA laddering, expression of genes related to apoptosis, and TUNEL assay. Similarly, the effect of homoeopathic medicines on apoptosis was measured by microarray analysis. Activity of Ruta 200C was compared with that of the mother tincture.
Ruta 200C produced morphological changes in the Dalton's lymphoma ascites tumor cells and induced DNA laddering. Carcinosinum 200C increased apoptotic gene p53 and Ruta 200C decreased antiapoptotic gene Bcl2. Administration of potentiated homoeopathic drugs to tumor-bearing mice induced TUNEL-positive cells in the tumor, showing increased apoptosis of tumor cells. Microarray analysis of cells treated with homoeopathic drugs indicated that many enzymes related to apoptosis were increased by homoeopathic drugs.
These data indicate that apoptosis is one of the mechanisms of tumor reduction of homeopathic drugs. A comparison of potentiated drugs with their mother tincture indicated that the potentiated drugs have biological activity similar to that of their mother tincture in spite of ultradilution.
The effects of Arnica montana 6cH on the individual modulation of acute inflammation kinetics in rats were evaluated. Adult male Wistar rats were inoculated with 1% carrageenan into the footpad and treated with Arnica montana 6cH, dexamethasone (4.0 mg/kg; positive control) or 5% hydroalcoholic solution (negative control), per os, each 15 minutes, between 30 and 180 minutes after the irritant inoculation. Histopathological and immunohistochemistry procedures were done in order to get a panel of inflammatory positive cells for CD3 (T lymphocytes), CD45RA (B lymphocytes), CD18 (beta 2 integrin), CD163 (ED2 protein), CD54 (ICAM-1), and MAC 387 (monocytes and macrophages). The statistical treatment of data included a posteriori classification of animals from each group (N = 20) in two subgroups presenting spontaneous precocious or late oedema. Animals that presented precocious oedema were less responsible to Arnica montana 6cH in relation to hemodynamic changes. Instead, rats that exhibited late oedema presented less intense oedema (P = .01), lower percentage of mast cell degranulation (P = .0001), and increase in lymphatic vessels diameter (P = .05). The data suggest an individually qualitative adjustment of inflammatory vascular events by Arnica montana 6cH.
Wound healing can be improved by transplanting mesenchymal stem cells (MSCs). In this study, we have demonstrated the benefits of the conditioned medium derived from human MSCs (CM-MSC) in wound healing using an excisional wound model. CM-MSC accelerated wound closure with increased
reepithelialization, cell infiltration, granulation formation, and angiogenesis. Notably, CM-MSC enhanced epithelial and endothelial cell migration, suggesting the contribution of increased cell migration to wound healing enhanced by CM-MSC. Cytokine array, ELISA analysis, and quantitative
RT-PCR revealed high levels of IL-6 in CM-MSC. Moreover, IL-6 added to the preconditioned medium enhanced both cell migration and wound healing, and antibodies against IL-6 blocked the increase in cell motility and wound closure by CM-MSC. The IL-6 secretory pathway of MSCs was inhibited by
SB203580, an inhibitor of p38 MAPK or siRNA against p38 MAPK, suggesting IL-6 secretion by MSCs is mediated through the activation of p38 MAPK. Inactivation of p38 MAPK also reduced the expression and production of IL-8 and CXCL1 by MSCs, both of which were also demonstrated to enhance cell
migration and wound closure. Thus, our data suggest MSCs promote wound healing through releasing a repertoire of paracrine factors via activation of p38 MAPK, and the CM-MSC may be applied to enhance wound healing.
Although cellular copper metabolism is tightly regulated through a variety of copper transport proteins and chaperones, disturbances in copper homeostasis are involved in several pathological disorders. The aim of this study was to evaluate the effects of extremely low copper concentrations on gene expression profiles of a line of human prostate epithelial cells (RWPE) which grows in the absence of fetal calf serum, a source of variable and unpredictable copper. Cells were exposed to copper(II) sulfate for 24h at concentrations varying from 10(-6) to 10(-17)M and untreated reference cells were exposed to the same volume of copper-free water. Relative gene expression variations between copper-treated and control cells were studied with microarray technology using the Whole Human Genome Array from Agilent. Microarray data demonstrated that copper added to the medium varied gene expression at all concentrations tested. Many genes belonging to functional gene families were modulated by copper, some dose-dependently. Amongst these genes metallothioneins (MT1A and MT2A) were over-expressed at all copper concentrations, MT1M was up-regulated between 10(-6) and 10(-9)M, while MT1B, MT1E, MT1G and MT1H were up-regulated between 10(-6) and 10(-14)M. The heat shock protein (HSP) gene family showed similar behavior: some HSP genes were constantly up-regulated by copper (HSP90Ad, HSP90B1 and HSPD1) and others only at higher concentrations (HSP90AB1 and HSPA8). Reverse-transcription-PCR analysis, performed on four different genes on five biological replicates for selected genes, on each copper concentration tested, confirmed the trend observed in microarray results. In conclusion, we unexpectedly observed a modulation of gene expression even at extremely diluted copper concentrations, similar to that induced by toxic concentrations, possibly as a result of very tight control of free copper(II) levels inside the cells.
Werner, Sabine, and Richard Grose. Regulation of Wound Healing by Growth Factors and Cytokines. Physiol Rev 83: 835–870, 2003; 10.1152/physrev.00032.2002.—Cutaneous wound healing is a complex process involving blood clotting, inflammation, new tissue formation, and finally tissue remodeling. It is well described at the histological level, but the genes that regulate skin repair have only partially been identified. Many experimental and clinical studies have demonstrated varied, but in most cases beneficial, effects of exogenous growth factors on the healing process. However, the roles played by endogenous growth factors have remained largely unclear. Initial approaches at addressing this question focused on the expression analysis of various growth factors, cytokines, and their receptors in different wound models, with first functional data being obtained by applying neutralizing antibodies to wounds. During the past few years, the availability of genetically modified mice has allowed elucidation of the function of various genes in the healing process, and these studies have shed light onto the role of growth factors, cytokines, and their downstream effectors in wound repair. This review summarizes the results of expression studies that have been performed in rodents, pigs, and humans to localize growth factors and their receptors in skin wounds. Most importantly, we also report on genetic studies addressing the functions of endogenous growth factors in the wound repair process.
Wound healing requires a complex series of reactions and interactions among cells and their mediators, resulting in an overlapping series of events including coagulation, inflammation, epithelialization, formation of granulation tissue, matrix and scar formation. Cytokines and chemokines promote inflammation, angiogenesis, facilitate the passage of leukocytes from circulation into the tissue, and contribute to the regulation of epithelialization. They integrate inflammatory events and reparative processes that are important for modulating wound healing. Thus both cytokines and chemokines are important targets for therapeutic intervention. The chemokine-mediated regulation of angiogenesis is highly sophisticated, fine tuned, and involves pro-angiogenic chemokines, including CXCL1-3, 5-8 and their receptors, CXCR1 and CXCR2. CXCL1 and CXCR2 are expressed in normal human epidermis and are further induced during the wound healing process of human burn wounds, especially during the inflammatory, epithelialization and angiogenic processes. Human skin explant studies also show CXCR2 is expressed in wounded keratinocytes and Th/1/Th2 cytokine modulation of CXCR2 expression correlates with proliferation of epidermal keratinocytes. Murine excision wound healing, chemical burn wounds and skin organ culture systems are valuable models for examining the role of inflammatory cytokines and chemokines in wound healing.