In vitro anti-proliferative, anti-bacterial potential and induction of DNA strand break of partially purified Cuscuta reflexa Roxb.
Abstract and Figures
Cuscuta reflexa is an important medicinal plant, mentioned in Ayurveda, an ancient Indian system of medicine. The plant is selected to evaluate the possibility for novel pharmaceuticals for anticancer and antibiotics drugs. Since most of these drugs had developed resisitance against currently used chemotherapeutics. This study describes the in vitro anti-proliferative, anti-bacterial and single stand DNA break of the holoprasitic plant Cuscuta reflexa. Bioassay-guided fractionation and partial purification of the plant were done and evaluated for antiproliferative activity against human cancer cell lines by SRB assay and single strand DNA break by comet assay. Further antibacterial activity was also performed by agar well diffusion assay. The alcoholic extract, chloroform fraction and partially purified ethylacetate-methanol (1:1) sub-fraction of C. reflexa showed anti-proliferative potential against IMR-32 and 502713 human cancer cell lines. Alcoholic extract exhibited anti-proliferative activity of 74% and 72%, chloroform fraction demonstrated 91% and 95% against neuroblastoma (IMR-32) and colon (502713) cancer cell lines at 100 μg/ml. Single strand DNA break of the chloroform fraction was also demonstrated using comet assay, indicating that possible mode of cell death may be apoptosis. Anti-microbial properties were evaluated against eight species of pathogenic and non-pathogenic microorganisms and maximum zone of inhibition for anti-bacterial activity was found against Staphylococcus aureus (22 mm) by alcoholic extract, 21 mm by chloroform fraction and 12 mm by ethylacetate-methanol (1:1) sub-fraction. Minimum inhibitory concentration (MIC) of the chloroform fraction was 1500 μg/ml for S. aureus. The plant was found to be equally effective against gram-positive and negative bacteria. Studies are well underway to isolate and identify active compounds from chloroform fraction and ethyl acetate:methanol (1:1) sub-fraction, which can be used as effective drug for various diseases.
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... Cuscuta reflexa (Giant dodder) in cell lines [5], Swiss albino mice [6], human red blood cells [7] and human cancer cell lines [8]. Ashwagandha or Indian ginseng Withania somnifera has immunomodulatory and anti-cancer effects [9][10][11][12][13][14]. Glycyrrhiza glabra (Liquorice) expresses promising antimicrobial, cytotoxic and anti-cancer effects [15,16]. ...
UNANI MEDICINE AND CANCER
Christer Sundqvist. Prepublished article, 2020
Petrafoundation, Helsinki, Finland https://www.petrafoundation.com/en/foundation
Unani medicine is an alternative medical system originating in ancient Greece almost 2500 years back. It is now practiced primarily in India. Herbal remedies, dietary practices and alternative therapies characterize Unani medicine. Let us study what it can offer for a cancer patient.
Ethnopharmacological relevance
Cuscuta reflexa Roxb. (C. reflexa) is a well-known traditional herbal plant, with numerous inherent therapeutic potentials including anticancer, antitumor, antibacterial, analgesic, anthelmintic, laxative and others. Moreover, the anticancer and antitumor potentials of this herb are ongoing with several trails, thus an attempt was made to assess the anticancer and hepatoprotective potentials of traditional C. reflexa herbs.
Method
The dried ethanolic extract of C. reflexa was tested for acute oral toxicity in the treated animals subsequently their behavioral, neurological, and autonomic profiles changes were observed. The preliminary anti-cancer effects of extracts against 1, 2- Dimethyl hydrazine (DMH) induced animals were assessed through barium enema X-ray, colonoscopy, and Aberrant crypt foci (ACF) studies. The blood samples of the animals (treated and untreated) were collected and their in-vitro histological parameters were evaluated by the experienced technician.
Results
It was observed that C. reflexa significantly reduced Disease activity indexing (DAI) level and ACF counting, as well as demonstrated similar activity as of the standard drug 5-Fluorouracil (5-FU). Histopathological results revealed that the apoptotic bodies decreased in the DMH-induced group (group II) during cancer progression while in 5-FU treated (group III) and C. reflexa treated (group IV and V) animals the apoptotic bodies were increased. Inversely, the mitotic bodies increased in group II animals and reduced in group III, IV, and V animals. In the colonic section, DMH-induced cancer assay exhibited significant effects on the levels of hemoglobin, Packed cell volume (PCV), Red blood cell (RBC) counts, Mean corpuscular hemoglobin concentration (MCHC), Mean corpuscular volume (MCV), and Mean cell hemoglobin (MCH), and was found to be less in group II animals whereas administration of C. reflexa efficiently recovered back the loss probably by healing the colon damage/depletion of cancer progression. Moreover, compared to the group II animals, the neutrophil count was within the normal range in C. reflexa administered group.
Conclusions
In the present study, the major hematological parameters significantly increased within DMH treated animals and exhibited extensive damage in the hepatic regions. Moreover, the histopathological findings demonstrated that the C. reflexa extracts potentially reduced the cell proliferation, with no toxicity. The C. reflexa extracts exhibited impending anti-cancer activity as well as protected the hepatic cells and thus could be potentially used in the management of colon or colorectal cancer and hepatic impairments.
According to traditional knowledge, Fagonia cretica has medicinal potential, especially against cancer and tumors. In the present study, this information was analyzed at laboratory level by performing cytotoxic, antitumor (potato disc) and DNA damage assay. Significant cytotoxic activity was found against brine shrimps at LD50 118.89 ppm, while antitumor assay showed that the extract inhibited tumor induction on potato discs. Significant antitumor activity was found against all the tumor-inducing Agrobacterium strains tested (At6, At10 and At77), with maximum tumor inhibition (77.04%) against At10. However, the extract did not show any lethal activity against Agrobacterium tumefaciens strains, and furthermore, no DNA damaging activity was observed. The overall results indicate a strong anti-cancerous potential of this plant.
When oxygen is metabolised or burned by the body cells, it forms by-product called free radicals. These free radicals or reactive oxygen species generated by oxidation is the main source of serious illness including many autoimmune diseases and ageing in the body. Antioxidant act as free radical terminators as it stops the free radical chain of oxidative reactive reactions by contributing hydrogen from phenolic hydroxyl group and themselves forming stable free radicals that do not initiate further oxidation. Thus, antioxidants which naturally occurring in plants has a potential protective functions against free radicals in the mechanism of preventing many chronic and acute diseases, which focused the attention of researcher. Poly-phenols and flavonoids are usually recognized as the medicinal plant compounds which are responsible for the antioxidant activity. The present paper views a new approach of disease caused by oxidative stress, potentiality of Indian traditional medicinal plant extracts and/or it's formulations against diseases through antioxidant activity and their health benefit.
The spread of multidrug-resistant (MDR) strains of bacteria necessitates the discovery of new classes of antibacterials and compounds that inhibit these resistance mechanisms. At present, there are no single chemical entity plant-derived antibacterials used clinically, and this chemically diverse group deserves consideration as a source for two major reasons. First, plants have exceptional ability to produce cytotoxic agents and second there is an ecological rationale that antimicrobial natural products should be present or synthesised de novo in plants following microbial attack to protect the producer from pathogenic microbes in its environment. We have been characterising plant-derived products that are either antibacterial in their own right, or modulators of resistance in bacterial strains possessing multidrug efflux mechanisms. These efflux transporters are responsible for resistance to certain antibiotics and antiseptics and occur in strains of methicillin-resistant Staphylococcus aureus (MRSA), a major clinical problem at present. We are also investigating plant sources for compounds with activity against mycobacteria with a view to discovering drug leads with potential activity toward tuberculosis (TB) producing species. This paper will briefly review the literature on plant derived bacterial resistance modifying agents and antibacterials. Examples in this area from our own work will be given. The activities of plant-derived antibacterials show that there are many potential new classes of antibacterial agents which should undergo further cytotoxicity, microbial specificity and preclinical studies.
Human lymphocytes were either exposed to X-irradiation (25 to 200 rads) or treated with H2O2 (9.1 to 291 μM) at 4 °C and the extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Both agents induced a significant increase in DNA migration, beginning at the lowest dose evaluated. Migration patterns were relatively homogeneous among cells exposed to X-rays but heterogeneous among cells treated with H2O2. An analysis of repair kinetics following exposure to 200 rads X-rays was conducted with lymphocytes obtained from three individuals. The bulk of the DNA repair occurred within the first 15 min, while all of the repair was essentially complete by 120 min after exposure. However, some cells demonstrated no repair during this incubation period while other cells demonstrated DNA migration patterns indicative of more damage than that induced by the initial irradiation with X-rays. This technique appears to be sensitive and useful for detecting damage and repair in single cells.
The in vitro antioxidant activity of Cuscuta reflexa stem extract has been investigated by estimating degree of non-enzymatic hemoglobin glycosylation measured colorimetrically at 440 nm. The ethyl acetate fraction of ethanol extract showed higher activity than the other fractions. The antioxidant activity of extracts are very close and identical in magnitude and comparable to that of standard antioxidant compounds used.
A precise regulation of redox balance is required for the cellular homeostatic control. Aberrant activation of redox-sensitive
transcription factors, such as nuclear factor-kappaB (NF-κB), activator protein 1 (AP-1), cyclic adenosine monophosphate response
element binding protein (CREB), and hypoxia inducible factor (HIF), contributes to carcinogenesis by promoting persistent
inflammation, abnormal cell proliferation, evasion from apoptosis, angiogenesis, etc. A wide variety of dietary phytochemicals
have been reported to exert cancer chemopreventive properties by suppressing the inappropriate activation of aforementioned
transcription factors. On the other hand, transcription of genes involved in the activation of cellular antioxidant arsenal
and carcinogen detoxification is largely regulated by another redox-sensitive transcription factor, i.e. NF-E2 related factor
2 (Nrf2), which plays a role in protecting cells/tissues from oxidative or electrophilic damage. Some food-derived phytochemicals
have been shown to activate Nrf2, thereby augmenting cellular antioxidant capacity and inducing expression of phase-2 detoxification
enzymes. Therefore, the modulation of cellular signaling mediated by redox-sensitive transcription factors in the right direction
represents a promising approach to achieving molecular target-based chemoprevention with edible phytochemicals.
Unlabelled:
Acacia catechu, commonly known as catechu, cachou and black cutch is an important medicinal plant and an economically important forest tree. The methanolic extract of this plant was found to have antimicrobial activities against six species of pathogenic and non-pathogenic microorganisms: Bacillus subtilis, Staphylococcus aureus, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The maximum zone of inhibition (20 mm) was found to be exhibited against S. aureus. For this organism the minimum bactericidal concentration (MBC) of the crude extract was 1,000 μg/ml. The extract was found to be equally effective against gram positive and gram negative bacteria. The antimicrobial activity of the extract was found to be decreased during purification. The chemical constituents of organic plant extracts were separated by thin layer chromatography (TLC) and the plant extracts were purified by column chromatography and were further identified by Gas chromatography-mass selection (GC-MS) analysis. The composition of A. catechu extract had shown major components of terpene i.e. camphor (76.40%) and phytol (27.56%) along with other terpenes in minor amounts which are related with their high antibacterial and antifungal properties.
Electronic supplementary material:
The online version of this article (doi:10.1007/s12088-011-0061-1) contains supplementary material, which is available to authorized users.