Article

Anticonvulsant activity of β-caryophyllene against pentylenetetrazol-induced seizures

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Abstract

Increasing evidence suggests that plant-derived extracts and their isolated components are useful for treatment of seizures and, hence, constitute a valuable source of new antiepileptic drugs with improved efficacy and better adverse effect profile. β-Caryophyllene is a natural bicyclic sesquiterpene that occurs in a wide range of plant species and displays a number of biological actions, including neuroprotective activity. In the present study, we tested the hypothesis that β-caryophyllene displays anticonvulsant effects. In addition, we investigated the effect of β-caryophyllene on behavioral parameters and on seizure-induced oxidative stress. Adult C57BL/6 mice received increasing doses of β-caryophyllene (0, 10, 30, or 100 mg/kg). After 60 min, we measured the latencies to myoclonic and generalized seizures induced by pentylenetetrazole (PTZ, 60 mg/kg). We found that β-caryophyllene increased the latency to myoclonic jerks induced by PTZ. This result was confirmed by electroencephalographic analysis. In a separate set of experiments, we found that mice treated with an anticonvulsant dose of β-caryophyllene (100 mg/kg) displayed an improved recognition index in the object recognition test. This effect was not accompanied by behavioral changes in the open-field, rotarod, or forced swim tests. Administration of an anticonvulsant dose of β-caryophyllene (100 mg/kg) did not prevent PTZ-induced oxidative stress (i.e., increase in the levels of thiobarbituric acid-reactive substances or the decrease in nonprotein thiols content). Altogether, the present data suggest that β-caryophyllene displays anticonvulsant activity against seizures induced by PTZ in mice. Since no adverse effects were observed in the same dose range of the anticonvulsant effect, β-caryophyllene should be further evaluated in future development of new anticonvulsant drugs.

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... Similarly to CBD, BCP is devoid of the psychoactive side effects associated with cannabinoid type-1 (CB 1 ) receptor activation, thus making BCP a novel candidate for a wide variety of therapeutic applications (Sharma et al., 2016;Gonçalves et al., 2020). This also includes seizures, as BCP has been shown to display antiepileptic effects in different animal models of epilepsy, as well as to attenuate inflammatory processes which are typically linked to hyperexcitability (Liu et al., 2015;de Oliveira et al., 2016;Tchekalarova et al., 2018). ...
... Previous studies have shown that BCP protects against seizures in preclinical models of epilepsy (Liu et al., 2015;de Oliveira et al., 2016;Tchekalarova et al., 2018). In our first approach (acute paradigm), we addressed the anticonvulsant effect of BCP against PTZ-induced seizures in both Syn1-Cre/Scn1a WT/A1783V and Scn1a WT/WT mice. ...
... The study was concentrated on two therapeutic benefits, on one hand, the capability of BCP to have antiseizuring activity in DS. This capability has been previously investigated in different animal models of epilepsy (e.g., models based on PTZ-or kainate-induced seizures), but not in DS, and has been related to its ability to attenuate inflammatory events which are typically linked to hyperexcitability (Liu et al., 2015;de Oliveira et al., 2016;Tchekalarova et al., 2018). Our data have corroborated the antiseizuring potential of BCP which attenuated the proconvulsant action of PTZ, with this potential being found in Syn1-Cre/Scn1a WT/A1783V mice but also in their controls (Scn1a WT/WT ) mice. ...
Article
Dravet Syndrome (DS) is caused by mutations in the Scn1a gene encoding the α1 subunit of the sodium channel Nav1.1, which results in febrile seizures that progress to severe tonic-clonic seizures and associated comorbidities. Treatment with cannabidiol has been approved for the management of seizures in DS patients, but it appears to be also active against associated comorbidities. In this new study, we have investigated β-caryophyllene (BCP), a cannabinoid with terpene structure that appears to also have a broad-spectrum profile, as a useful therapy against both seizuring activity and progression of associated comorbidities. This has been studied in heterozygous conditional knock-in mice carrying a missense mutation (A1783V) in Scn1a gene expressed exclusively in neurons of the Central Nervous System (Syn-Cre/Scn1aWT/A1783V), using two experimental approaches. In the first approach, an acute treatment with BCP was effective against seizuring activity induced by pentylenetetrazole (PTZ) in wildtype (Scn1aWT/WT) and also in Syn-Cre/Scn1aWT/A1783V mice, with these last animals having a greater susceptibility to PTZ. Such benefits were paralleled by a BCP-induced reduction in PTZ-induced reactive astrogliosis (labelled with GFAP) and microgliosis (labelled with Iba-1) in the prefrontal cortex and the hippocampal dentate gyrus, which were visible in both wildtype (Scn1aWT/WT) and Syn-Cre/Scn1aWT/A1783V mice. In the second approach, both genotypes were treated repeatedly with BCP to investigate its effects on several DS comorbidities. Thus, BCP corrected important behavioural abnormalities of Syn-Cre/Scn1aWT/A1783V mice (e.g. delayed appearance of hindlimb grasp reflex, induction of clasping response, motor hyperactivity, altered social interaction and memory impairment), attenuated weight loss, and slightly delayed premature mortality. Again, these benefits were paralleled by a BCP-induced reduction in reactive astrogliosis and microgliosis in the prefrontal cortex and the hippocampal dentate gyrus typical of Syn-Cre/Scn1aWT/A1783V mice. In conclusion, BCP was active in Syn-Cre/Scn1aWT/A1783V mice against seizuring activity (acute treatment) and against several comorbidities (repeated treatment), in both cases in association with its capability to reduce glial reactivity in areas related to these behavioural abnormalities. This situates BCP in a promising position for further preclinical evaluation towards a close translation to DS patients.
... Preclinical tests with BCP were previously conducted with excipient solutions consisting of olive oil [14], DMSO [15], aqueous solution with Tween 80 [16], among others. These formulations do not provide substantial protection against oxidation of the active. ...
... This demonstrates the recent increase in interest about this compound. Most of these articles address the new therapeutic functions and mechanisms of action proposed for BCP. Figure 2 shows the main therapeutic applications evidenced for BCP in preclinical studies [11,[14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31]. There is a consensus in the scientific literature that immuno-inflammatory changes and oxidative stress are common factors in the pathogenesis of various diseases [32][33][34][35]. ...
Article
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Abstract: Background: The β-caryophyllene (BCP), a phytocannabinoid present in various essential oils, demonstrated selective action on the CB2 endocannabinoid receptor and attracted considerable attention because of its several pharmacological activities. Despite this recognized potential, this hydrophobic compound is a volatile and acid-sensitive sesquiterpene that readily oxidizes when exposed to air, and has low bioavailability in oral formulations. Thus, the development of formulations that guarantee its stability and increase its bioavailability is a challenge for its use in the pharmaceutical field. Methods: The present review brings for the first time a comprehensive overview of the controlled and vectorized release formulations tested for BCP administration. Among these, we have addressed nanoemulsions, inclusion complexes with cyclodextrins, liposomes, wound dressings, nanocomposites and nanoparticles. A literature search was performed on Pubmed, Web of Science and Science direct, and patents documents were also searched on European Patent Office, World Intellectual Property Organization and Brazilian National Institute of Industrial Property. Results: The systems presented here may represent an interesting approach to overcome the limitations already mentioned for this terpene. These systems proved to be promising for improving solubility, stability and controlled release of this pharmacological relevant sesquiterpene. In the industrial field, some companies have filed patent applications for the commercial use of the BCP, however, the use of pharmaceutical formulations still appeared moderate. Conclusion: This prospective study evidenced the new perspectives related to BCP vectorization systems in the pharmaceutical and industrial marketing field and may serve as a basis for further research and pharmaceutical use of this powerful cannabinoid.
... The lines of the apparatus was divided the floor into sixteen 18 x 18 cm squares and the central square (18 x 18 cm) was drawn in the middle of cage (The middle square is used to test the mouse locomotors activity and their behaviour crossing the lines of the trailing chamber many times during a test session). The increase in the count of mice movement was regarded to central nervous stimulation while a decrease in the number of mice movement was regarded to central nervous depressant activity [19]. ...
... With regards to mortality rate, there is no statistically difference between PTZ and extract dose 1 (100 mg) groups, while extract dose 2 (200 mg) and Valproate groups hindered the mortality among mice. The open field routinely used to test gross animal behaviour, the nerve excitability, anxiety, mental activity, exploration, and locomotion [19]. However, the epileptic patient has shown to develop different psychotic impairments like anxiety and showed to be less mental activity comparing them with normal individuals. ...
... Kaempferol Anti-apoptotic, pro-wound healing, anti-cancer, cardioprotective, anti-oxidant, pro-apoptotic, anti-allergic, anti-parasitic, antidiabetic, anti-adipogenic, anti-thrombotic, anti-inflammatory, anti-metabolic syndrome, anti-bacterial, immunoregulatory, hepatoprotective, anti-atherosclerosis [28][29][30][31][32][33][34][35] Linalool Anti-parasitic, anti-convulsant, anti-cancer, anti-bacterial, neuroprotective, anti-oxidant, anti-inflammatory, anti-Alzheimer, anxiolytic, hepatoprotective, anti-hyperalgesic, neuroprotective [36][37][38][39][40][41][42][43] Citronellal Anti-fungal, insect repellant, hepatoprotective, anti-nociceptive, anti-inflammatory, anti-bacterial [44][45][46] Caryophyllene Anti-cancer, anti-mutagenic, anti-bacterial, oxygen deprivation protective, neuroprotective, hepatoprotective, anti-convulsant, anti-diabetic, anti-microbial, anti-Alzheimer, pro-longevity, analgesic, nephroprotective [47][48][49][50][51][52][53][54][55][56] Humulene Insecticidal, anti-cancer, anti-inflammatory [51,57,58] Neridiol Anti-parasitic, antioxidant, neuroprotective, pro-wound healing, anti-microbial [59][60][61][62] While it is clear that L. candidum possesses many valuable compounds with considerable therapeutic potential, to the best of our knowledge, no bioinformatical research has been carried out to assess their medicinal potential. In order to close this gap, we analyzed the potential human therapeutic targets (proteins and other biomolecules) of the above-mentioned compounds, using the STITCH database (http://stitch.embl.de/) ...
... Kaempferol Anti-apoptotic, pro-wound healing, anti-cancer, cardioprotective, anti-oxidant, pro-apoptotic, anti-allergic, anti-parasitic, anti-diabetic, anti-adipogenic, anti-thrombotic, anti-inflammatory, anti-metabolic syndrome, anti-bacterial, immunoregulatory, hepatoprotective, anti-atherosclerosis [28][29][30][31][32][33][34][35] Linalool Anti-parasitic, anti-convulsant, anti-cancer, anti-bacterial, neuroprotective, anti-oxidant, anti-inflammatory, anti-Alzheimer, anxiolytic, hepatoprotective, anti-hyperalgesic, neuroprotective [36][37][38][39][40][41][42][43] Citronellal Anti-fungal, insect repellant, hepatoprotective, anti-nociceptive, anti-inflammatory, anti-bacterial [44][45][46] Caryophyllene Anti-cancer, anti-mutagenic, anti-bacterial, oxygen deprivation protective, neuroprotective, hepatoprotective, anti-convulsant, anti-diabetic, anti-microbial, anti-Alzheimer, pro-longevity, analgesic, nephroprotective [47][48][49][50][51][52][53][54][55][56] Humulene Insecticidal, anti-cancer, anti-inflammatory [51,57,58] Neridiol Anti-parasitic, antioxidant, neuroprotective, pro-wound healing, anti-microbial [59][60][61][62] While it is clear that L. candidum possesses many valuable compounds with considerable therapeutic potential, to the best of our knowledge, no bioinformatical research has been carried out to assess their medicinal potential. In order to close this gap, we analyzed the potential human therapeutic targets (proteins and other biomolecules) of the above-mentioned compounds, using the STITCH database (http://stitch.embl.de/) ...
Article
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Lilium candidum L., known as Madonna, meadow, or white lily, is a bulbous plant from the Liliaceae family, originating in the Middle East. L. candidum has been abundantly used in folk medicine since ancient times to relieve a variety of ailments, including age-related diseases, burns, ulcers, and coughs. The aim of this article is to investigate the anti-inflammatory and anti-diabetic activities of L. candidum extracts and its active phytochemicals. Some active volatile phytochemicals were identified using gas chromatography–mass spectrometry (GC-MS) analysis. Significant (p < 0.001) anti-diabetic properties of the extracts kaempferol, linalool, citronellal, and humulene were demonstrated by an elevation in glucose uptake by adipocytes. The significant (p < 0.01) effect of the plant extracts kaempferol, citronellal, and humulene on the secretion of pro-inflammatory cytokines interleukin 6 (IL-6) and interleukin 8 (IL-8) was demonstrated using enzyme-linked immunosorbent assay. Altogether, L. candidum and its rich collection of phytochemicals hold promising medicinal potential, and further investigations of its therapeutic prospects are encouraged.
... This result agrees with Shaw et al. [31] who suggested that Long-Evans rats with epilepsy had increased immobility time in the forced swim test (FST). Our result was also in tandem with that of [32] who observed in their pentylenetetrazole model of epilepsy that epileptic rats showed sign of despair by an increased immobility time within the FST. Interestingly, in this study, we found that the history of PFS did not exacerbate immobility in the forced swim test; this may be due to the transient effect of PFS in the animals. ...
Article
Background Depression is the most significant cause of suicide among neuropsychiatric illnesses. Major depression further affects the quality of life in an individual with epilepsy. The treatment of depression in an epileptic patient could be very challenging because of drug selection or the fact that some antiepileptic drugs are known to cause depression. It has been shown that in addition to the known involvement of the serotonergic pathway in depression, the glutamatergic system is also involved in the evolution of the disease, but this knowledge is limited. This study assessed if induction of epilepsy in rats will cause depressive-like behavior, alters the concentrations of metabotropic receptor 5 (mGluR5), glutamate transport protein (GLAST), glutamate synthase (GS) and brain derived neurotrophic factor (BDNF). Materials and method Epilepsy was induced in rats by injecting Pentylenetetrazole at 35 mg/kg every other day. At kindle, rats were subjected to sucrose preference test (SPT) and forced swim test (FST) and decapitated 4 h later. Hippocampal tissue was collected and the BDNF concentration was measured with ELISA; mGluR5 and GS protein expression was measured using western blot while amygdala tissue was used for GLAST expression with flow cytometry. Results Our results showed that epilepsy leads to depressive-like behavior in rats and alters the glutamatergic system. Conclusion Therefore, we conclude that targeting the glutamate pathway may be a good strategy to alleviate depressive-like behavior associated with epilepsy.
... In addition, these variations could be attributed to endogenous factors such as individual genetic variability [32]. β-Caryophyllene is reported to possess anticancer, analgesic [34], anticonvulsant [35], anti-inflammatory [36], antioxidant, and antimicrobial activities [37], and its abundance, especially in the E, EQ, and ESEG varieties, makes them a possible source of this compounds. As a result, β-caryophyllene was described as a potential agent to treat several diseases due to its activity [38]. ...
Article
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Guava (Psidium guajava) leaves are commonly used in the treatment of diseases. They are considered a waste product resulting from guava cultivation. The leaves are very rich in essential oils (EOs) and volatiles. This work represents the detailed comparative chemical profiles of EOs derived from the leaves of six guava varieties cultivated in Egypt, including Red Malaysian (RM), El-Qanater (EQ), White Indian (WI), Early (E), El-Sabahya El-Gedida (ESEG), and Red Indian (RI), cultivated on the same farm in Egypt. The EOs from the leaves of guava varieties were extracted by hydro-distillation and analyzed with GC-MS. The EOs were categorized in a holistic manner using chemometric tools. The hydro-distillation of the samples yielded 0.11–0.48% of the EO (v/w).The GC-MS analysis of the extracted EOs showed the presence of 38 identified compounds from the six varieties. The sesquiterpene compounds were recorded as main compounds of E, EQ, ESEG, RI, and WI varieties, while the RM variety attained the highest content of monoterpenes (56.87%). The sesquiterpenes, β-caryophyllene (11.21–43.20%), and globulol (76.17–26.42%) were detected as the major compounds of all studied guava varieties, while trans-nerolidol (0.53–10.14) was reported as a plentiful compound in all of the varieties except for the RM variety. A high concentration of D-limonene was detected in the EOs of the RM (33.96%), WI (27.04%), and ESEG (9.10%) varieties. These major compounds were consistent with those reported for other genotypes from different countries. Overall, the EOs’ composition and the chemometric analysis revealed substantial variations among the studied varieties that might be ascribed to genetic variability, considering the stability of the cultivation and climate conditions. Therefore, this chemical polymorphism of the studied varieties supports that these varieties could be considered as genotypes of P. guajava. It is worth mentioning here that the EOs, derived from leaves considered to be agricultural waste, of the studied varieties showed that they are rich in biologically active compounds, particularly β-caryophyllene, transnerolidol, globulol, and D-limonene. These could be considered as added value for pharmacological and industrial applications. Further study is recommended to confirm the chemical variations of the studied varieties at a molecular level, as well as their possible medicinal and industrial uses.
... Five tests were performed in this study for the assessment of cognition impairment, recognition memory, anxiety and behavioral normalcy, motor coordination impairment, and spatial memory retention. Utilization of multiple behavioral screening tests in epilepsy studies for the assessment of various behavioral parameters has been reported previously [42][43][44][45][46][47][48]. The HMPs were then assessed for their effect on brain oxidative stress through the measurement of malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) levels. ...
Article
Epileptic seizures are characterized by imbalanced inhibition-excitation cycle that triggers biochemical alterations responsible for jeopardized neuronal integrity. Conventional antiepileptic drugs (AEDs) have been the mainstay option for treatment and control; however, symptomatic control and potential to exacerbate the seizure condition calls for viable alternative to these chemical agents. In this context, natural product-based therapies have accrued great interest in recent years due to competent disease management potential and lower associated adversities. Cicuta virosa (CV) is one such herbal remedy that is used in traditional system of medicine against myriad of disorders including epilepsy. Homeopathic medicinal preparations (HMPs) of CV were assessed for their efficacy in pentylenetetrazole (PTZ)-induced acute and kindling models of epilepsy. CV HMPs increased the latency and reduced the duration of tonic-clonic phase in acute model while lowering the kindling score in the kindling model that signified their role in modulating GABAergic neurotransmission and potassium conductance. Kindling-induced impairment of cognition, memory, and motor coordination was ameliorated by the CV HMPs that substantiated their efficacy in imparting sustained neuronal fortification. Furthermore, biochemical evaluation showed attenuated oxidative stress load through reduced lipid peroxidation and strengthened free radical scavenging mechanism. Taken together, CV HMPs exhibited promising results in acute and kindling models and must be further assessed through molecular and epigenomic studies.
... for 30 days) significantly decreased cauda epididymal sperm count, percentages of both motile, and with normal morphology sperm. Nevertheless, it neither affected the ultrastructure of the cauda epididymis or testes nor the histology of the treated rats (Al-Alami et al. 2015) as shown in Table 3. Due to its safe contraceptive dose for both male sexual and other organs and its selectivity towards antisperm action (post-testicular targeting), in addition to its reversibility (Correia et al. 2016;Tambe et al. 1996;Al-Alami et al. 2015), it may be considered as a promising natural male contraceptive (see Tables 3, 4, 5). Furthermore, it Phytochem Rev should be evaluated for 60 days instead of 30 days (complete spermatogenesis cycle of rats is from 48 to 60 days) by oral administration after enhancement of its poor oral bioavailability to accurately judge its safety and efficacy as shown in Table 5 (Fidyt et al. 2016;Di Sotto et al. 2018;Zeng et al. 2006). ...
Article
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The use of hazardous female contraceptives is still the first choice in family planning programs. Moreover, all the drugs examined to be used as infertility inducers in males or as spermicides into the cervix were not sufficiently efficient, in addition to not having complete safety. Therefore, many researchers were directed to assess complementary and alternative medicine to address these harmful effects. Accordingly, this review aimed to evaluate the efficacy of natural products alternatives in the induction of male infertility (male contraception) and/or proceeding of spermicidal action (vaginal contraception), although, that concept may be contradictory or prohibited in some developing and well-developed countries due to low birth rate problems, traditional, and religious beliefs. This study covers the literature from 1968 to 2019. In this review, we included previous controlled experiments with adult rats, mice, rabbits, monkeys and humans with clearly complete experimental criteria. The interventions were natural products derived from raw or refined plants, parts of plants or semi-synthesized compounds. The additional data such as toxicity and safety profiles, irreversibility, mechanism of actions, microbicidal activity, effective doses or concentrations, quantitative analysis in daily food and oral bioavailability, were included to identify the most promising natural products for contraception. In accordance with vaginal contraception, there were some herbal extracts and/or principles that may be used as prospective candidates after performing more pre-clinical and clinical trials. These products are acaciaside-B, Achyranthes aspera protein, xanthotoxin, oleanolic acid 3-β-d-glucuronide, ursolic acid, bivittoside D, subtilosin, Sapindus mukorossi saponins and Sapindus saponaria saponins fractions. Moreover, some products may be considered as effective and safe male contraceptives after clinical evaluation; which are NB-DNJ, NB-DGJ, β-caryophyllene, oleanolic acid, Tripterygium wilfordii glycosides, tripchlorolide, Carica papaya fatty acid esters and ZNF185-derived peptide. Generally, natural products that have high potential to be used as drug candidates concerning male and vaginal contraception are limited.
... BCP was found to display anticonvulsant activity against seizures induced by pentylenetetrazole in mice. Since no adverse effects were observed when BCP was administered at the concentration of 100 mg/kg b.w., and because of the lack of genotoxicity [178], this compound was considered a potential new anticonvulsant drugs [179]. Moreover, BCP was clinically useful as an adjunct treatment against seizure spread and status epilepticus and concomitant oxidative stress, neurotoxicity and cognitive impairments [180]. ...
Article
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E)-β-caryophyllene (BCP) is a bicyclic sesquiterpene widely distributed in the plant kingdom, where it contributes a unique aroma to essential oils and has a pivotal role in the survival and evolution of higher plants. Recent studies provided evidence for protective roles of BCP in animal cells, highlighting its possible use as a novel therapeutic tool. Experimental results show the ability of BCP to reduce pro-inflammatory mediators such as tumor necrosis factor-alfa (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), thus ameliorating chronic pathologies characterized by inflammation and oxidative stress, in particular metabolic and neurological diseases. Through the binding to CB2 cannabinoid receptors and the interaction with members of the family of peroxisome proliferator-activated receptors (PPARs), BCP shows beneficial effects on obesity, non-alcoholic fatty liver disease/nonalcoholic steatohepatitis (NAFLD/NASH) liver diseases, diabetes, cardiovascular diseases, pain and other nervous system disorders. This review describes the current knowledge on the biosynthesis and natural sources of BCP, and reviews its role and mechanisms of action in different inflammation-related metabolic and neurologic disorders.
... The study was performed in accordance with the guidelines for the care and use of laboratory animals approved by the Local Ethical Committee of Eskisehir Osmangazi University (25.06.2019/745). PTZ, diazepam, NaHS, CORM-2, LARG and L-NAME were dissolved in saline and PTZ injected 30 min after the last treatment with all agents according to de Oliveira et al. (18). A known anxiolytic drug, diazepam, was used as a positive control. ...
Article
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Objective: The present study aimed to investigate the anticonvulsant effects of bioactive gas mediator’s hydrogen sulfide (H2S), carbon monoxide (CO) and nitric oxide (NO) in pentylenetetrazole (PTZ)-induced convulsions in rats. Material and Methods: Eighty male Wistar-Albino rats randomly divided into ten groups. Convulsions were induced by administering 60 mg/kg PTZ intraperitonally (i.p). Diazepam (1 and 2 mg/kg), sodium hydrosulfide (NaHS, 5 and 10 mg/kg), CO releasing molecule-2 (CORM-2, 5 and 10 mg/kg), L-arginine (LARG, 30 and 100 mg/kg) and L-NG-Nitro arginine methyl ester (L-NAME, 30 mg/kg) were administered 30 minutes after PTZ. Seizure latency, seizure duration and seizure score were evaluated. Data were analyzed by the Kruskal Wallis test followed by posthoc analyses with Tukey. Any p value <0.05 was considered statistically significant. Results: NaHS (10 mg/kg) significantly reduced seizure duration and seizure in comparison with PTZ similar to diazepam (2 mg/kg), but it did not cause any significant change in seizure latency. Other agents did not significantly change seizure latency, seizure duration, and seizure score compared to PTZ. Conclusion: NaHS may be protective in PTZ-induced seizures. More research is needed to investigate the mechanisms underlying this effect.
... PTZ is a chemical substance traditionally used in rodent models to induce seizures [34,35], as well as to investigate the neurobiology mechanisms of epilepsy [36,37]. Both larval and adult zebrafish are pharmacologically susceptible to convulsing agents (i.e., kainic acid and PTZ) [17,[38][39][40][41]. ...
Article
Epilepsy is characterized by abnormal and recurrent hyperexcitability in brain cells. Various comorbidities are associated with epilepsy, including irritability and aggressive behavior. Aggression is a negative effect observed in epileptic patients that may be harmful to other individuals, impairing social relations. Thus, developing novel experimental models to assess behavioral phenotypes that may comorbid with neurological disorders are of great interest. Here, we investigate whether pentylenetetrazole (PTZ) increases aggression in zebrafish following a single PTZ exposure. Animals were exposed to 10 mM PTZ for 20 min and aggression-towards mirror was measured at different time intervals after recovering period (1 h, 3 h, 6 h, 24 h, 48 h, and 72 h). We observed that zebrafish showed exacerbated aggression, as well as an increased number of entries in the virtual conspecific area from 1 h to 48 h after PTZ. However, no behavioral differences were observed after 72 h. Overall, our novel findings show that PTZ evokes aggression after a single PTZ exposure in a time-dependent manner, reinforcing the use of zebrafish models to explore epilepsy-related comorbidities.
... β-Caryophyllene is a major volatile organic bicyclic sesquiterpene that has traditionally been used to provide a woody, spicy aroma and fragrance to cosmetics and perfumes. Studies have suggested that β-caryophyllene also possesses anti-inflammatory (Andrade-Silva et al., 2016), antioxidant (Basha & Sankaranarayanan, 2016), anticonvulsant (Oliveira et al., 2016), anti-alcoholism (Bahi et al., 2014), antinociceptive (Katsuyama et al., 2013), anxiolytic, antidepressant (Bahi et al., 2014), antiallodynic (Segat et al., 2017), cardioprotective, hepatoprotective, gastroprotective, nephroprotective (Machado et al., 2018) and anticarcinogenic (Dahham et al., 2015) activities. ...
Article
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The sesquiterpene β-caryophyllene is an ubiquitous component in many plants that has commercially been used as an aroma in cosmetics and perfumes. Recent studies have shown its potential use as a therapeutic agent and biofuel. Currently, β-caryophyllene is isolated from large amounts of plant material. Molecular farming based on the Nicotiana benthamiana transient expression system may be used for a more sustainable production of β-caryophyllene. In this study, a full-length cDNA of a new duplicated β-caryophyllene synthase from Artemisia annua (AaCPS1) was isolated and functionally characterized. In order to produce β-caryophyllene in vitro, the AaCPS1 was cloned into a plant viral-based vector pEAQ-HT. Subsequently, the plasmid was transferred into the Agrobacterium and agroinfiltrated into N. benthamiana leaves. The AaCPS1 expression was analyzed by quantitative PCR at different time points after agroinfiltration. The highest level of transcripts was observed at 9 days post infiltration (dpi). The AaCPS1 protein was extracted from the leaves at 9 dpi and purified by cobalt-nitrilotriacetate (Co-NTA) affinity chromatography using histidine tag with a yield of 89 mg kg −1 fresh weight of leaves. The protein expression of AaCPS1 was also confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analyses. AaCPS1 protein uses farnesyl diphosphate (FPP) as a substrate to produce β-caryophyllene. Product identification and determination of the activity of purified AaCPS1 were done by gas chromatography-mass spectrometry (GC-MS). GC-MS results revealed that the AaCPS1 produced maximum 26.5 ± 1 mg of β-caryophyllene per kilogram fresh weight of leaves after assaying with FPP for 6 h. Using AaCPS1 as a proof of concept, we demonstrate that N. benthamiana can be considered as an expression system for production of plant proteins that catalyze the formation of valuable chemicals for industrial applications.
... In a neurovascular unit model of oxygen-glucose deprivation and reoxygenation-induced injury, β-caryophyllene significantly decreased blood-brain barrier (BBB) permeability, reduced neuronal apoptosis, relieved oxidative stress damage, decreased secretion of inflammatory cytokines, downregulated metalloproteinase-9 expression/activity and Bcl-2-associated X protein (Bax) expression, and upregulated expression of claudin-5, occludin, zonula occludens-1 (ZO-1), growth-associated protein-43 (GAP-43) and B-cell lymphoma 2 (Bcl-2) [155]. Conversely, β-caryophyllene relieved seizures in mice induced by pentylenetetrazole, but anti-convulsant doses (0, 10, 30, and 100 mg/kg i.p.) showed no benefits over pentylenetetrazole related oxidative stress i.e., thiobarbituric acid-reactive substances and nonprotein thiol content [156]. Lou et al. (2016) found an attenuation of focal cerebral ischemia-reperfusion injury in rats by treatment with β-caryophyllene through enhanced expression of Nrf2 and HO-1, and restored activity and expression of antioxidant enzymes, i.e., superoxide dismutase (SOD) and catalase (CAT) [157]. ...
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Mitochondrial dysfunction results in a series of defective cellular events, including decreased adenosine triphosphate (ATP) production, enhanced reactive oxygen species (ROS) output, and altered proteastasis and cellular quality control. An enhanced output of ROS may damage mitochondrial components, such as mitochondrial DNA and elements of the electron transport chain, resulting in the loss of proper electrochemical gradient across the mitochondrial inner membrane and an ensuing shutdown of mitochondrial energy production. Neurons have an increased demand for ATP and oxygen, and thus are more prone to damage induced by mitochondrial dysfunction. Mitochondrial dysfunction, damaged electron transport chains, altered membrane permeability and Ca2+ homeostasis, and impaired mitochondrial defense systems induced by oxidative stress, are pathological changes involved in neurodegenerative disorders. A growing body of evidence suggests that the use of antioxidants could stabilize mitochondria and thus may be suitable for preventing neuronal loss. Numerous natural products exhibit the potential to counter oxidative stress and mitochondrial dysfunction; however, science is still looking for a breakthrough in the treatment of neurodegenerative disorders. β-caryophyllene is a bicyclic sesquiterpene, and an active principle of essential oils derived from a large number of spices and food plants. As a selective cannabinoid receptor 2 (CB2) agonist, several studies have reported it as possessing numerous pharmacological activities such as antibacterial (e.g., Helicobacter pylori), antioxidant, anti-inflammatory, analgesic (e.g., neuropathic pain), anti-neurodegenerative and anticancer properties. The present review mainly focuses on the potential of β-caryophyllene in reducing oxidative stress and mitochondrial dysfunction, and its possible links with neuroprotection.
... However, β-caryophyllene from essential oils has not been reported for the treatment of H. pylori infection. β-caryophyllene has been known to have anti-inflammatory effects and has showed efficacy with respect to the management of degenerative brain diseases such as depression, cerebral ischemic injury, Alzheimer's disease, and epilepsy [33][34][35][36][37]. In addition, β-caryophyllene is effective against lymphoma and neuroblastoma because of its anti-inflammatory effects [38]. ...
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New antibacterial treatments against Helicobacter pylori are needed as H. pylori is acquiring antibiotic resistance. β-caryophyllene is a natural bicyclic sesquiterpene, with anti-inflammatory and antimicrobial effects. This study investigates the effects of H-002119-00-001 from β-caryophyllene on the eradication of H. pylori in a mouse model, and its effects on the inflammation of the gastric mucosa. To evaluate the anti-H.pylori efficacy of β-caryophyllene, a total of 160 mice were divided into eight groups (n = 10 each) and were administered different treatments for 2 and 4 weeks. H. pylori eradication was assessed using a Campylobacter-like organism (CLO) test and H. pylori qPCR of the gastric mucosa. The levels of inflammation of gastric mucosa were assessed using histology and immunostaining. H-002119-00-001 decreased bacterial burden in vitro. When H-002119-00-001 was administered to mice once daily for 2 weeks, cure rates shown by the CLO test were 40.0%, 60.0%, and 70.0% in groups 6, 7, and 8, respectively. H. pylori levels in gastric mucosa decreased dose-dependently after H-002119-00-001 treatment. H-002119-00-001 also reduced levels of inflammation in gastric mucosa. H-002119-00-001 improved inflammation and decreased bacterial burden in H. pylori-infected mouse models. H-002119-00-001 is a promising and effective therapeutic agent for the treatment of H. pylori infection.
... In the pentilenotetrazole-induced seizure model, β-caryophyllene (100 mg/kg, i.p.) increased the latency to myoclonic seizures as compared to the control group as confirmed using electroencephalogram, revealing that such changes also occur at electrophysiological levels. It is worth noting that β-caryophyllene at the dose in which it presented anticonvulsant activity improved the recognition rate for the object in the recognition test, without motor alterations in the open field, Rota-rod and forced-swim tests [143]. However, different from what was observed by Liu et al. [136], β-caryophyllene did not prevent oxidative stress as induced by pentylenetetrazole [136]. ...
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Epilepsy is a most disabling neurological disorder affecting all age groups. Among the various mechanisms that may result in epilepsy, neuronal hyperexcitability and oxidative injury produced by an excessive formation of free radicals may play a role in the development of this pathology. Therefore, new treatment approaches are needed to address resistant conditions that do not respond fully to current antiepileptic drugs. This paper reviews studies on the anticonvulsant activities of essential oils and their chemical constituents. Data from studies published from January 2011 to December 2018 was selected from the PubMed database for examination. The bioactivity of 19 essential oils and 16 constituents is described. Apiaceae and Lamiaceae were the most promising botanical families due to the largest number of reports about plant species from these families that produce anticonvulsant essential oils. Among the evaluated compounds, β-caryophyllene, borneol, eugenol and nerolidol were the constituents that presented antioxidant properties related to anticonvulsant action. These data show the potential of these natural products as health promoting agents and use against various types of seizure disorders. Their properties on oxidative stress may contribute to the control of this neurological condition. However, further studies on the toxicological profile and mechanism of action of essential oils are needed.
... 16,17 In animal works, one of the widely used compounds to induce seizure is PTZ. 18 In a similar manner with previous studies, 6 in our study injection of PTZ was associated with the development of seizure attacks in mice. ...
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Background: Frequent seizure is followed by overproduction of free radicals and brain oxidative stress. Renin angiotensin system (RAS) has some effects on central nervous system. We designed this research to challenge the effect of captopril as an angiotensin converting enzyme (ACE) inhibitor against brain oxidative stress in pentylenetetrazole (PTZ) -induced seizures in mice. Methods: The groups were including (1) Control (saline); (2) PTZ (100 mg/kg, i.p.), (3-5) PTZ- captopril (Capto) that received three doses of Capto 10, 50 and 100 mg/kg 30 min before PTZ injection. Latency time in the onset minimal clonic seizures (MCS) and generalized tonic-clonic seizures (GTCS) were recorded. The level of malondialdehyde (MDA) and total thiol, as well as superoxide dismutase (SOD) and catalase (CAT) activity in the hippocampus and cortex were measured. Results: All doses of captopril postponed the onset of MCS and GTCS. Accumulation of MDA in the brain tissues of PTZ group was higher than control group, while total thiol content and CAT activity were lower. Pretreatment with captopril (100 mg/kg) diminished MDA concentration compared with PTZ group. Captopril (50 and 100 mg/kg) also increased the level of total thiol groups versus PTZ group. Captopril injection (50 and 100 mg/kg) elevated the activity of SOD and CAT in the brain tissues. In addition captopril administration diminished mortality rate caused by PTZ. Conclusion: Findings demonstrated that convulsions caused by PTZ were followed by oxidative stress status in the brain tissues. Pretreatment with captopril attenuated the effect of PTZ on brain tissue oxidative damage.<br /
... The PTZ group (n = 10): the rats received 0.05% CMC one week before an s.c. injection with PTZ (60 mg/kg b.w.), according to de Oliveira et al. [23]. PTZ at a dose of 60 mg/kg b.w. was preferred as higher doses (70 mg and more) can cause more rapid death of rats. ...
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Introduction: Neuronal cell death and glial cell activation are the main pathological findings induced by seizures secondary to oxidative stress. Previous studies have explained neuronal cell death on the basis of cell necrosis and apoptosis. Recent studies have attributed the neuronal loss to autophagy. The proved antioxidant and antifibrotic effect of nilotinib favours its use in the management of epileptic seizures. Aim of the study: was to analyse the neuroprotective and antiepileptic effect of nilotinib and explain its mechanism of action. Material and methods: Forty adult male rats were divided into four groups: control, pentylenetetrazol (PTZ) group (injected with PTZ 60 mg/kg, s.c.), pregabalin (Pregb)-PTZ group (pretreated with pregabalin daily 30 mg/kg; orally for 1 week) and nilotinib (NIL)-PTZ group (pretreated with nilotinib, 25 mg/kg daily for 1 week) prior to PTZ. Seizure latency was evaluated, the hippocampus tissue level of antioxidant enzymes was assessed. The histopathological changes in the hippocampus were studied using hematoxylin and eosin stain and immunohistochemical stain for brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), beclin-1, nuclear factor kappa-B (NF-κB) and Bcl-2-like protein 4 (BAX). Results: Nilotinib induced an increase in the latency of seizures, enhanced the antioxidant levels of the γ-aminobutyric acid and nuclear factor (erythroid-derived 2)-like 2 activities together with the improvement of the hippocampal histology. A reduction was reported for BDNF, GFAP, beclin-1, NF-κB and BAX expression in nerve cells. Conclusions: Nilotinib may have promising neuroprotective and antiepileptic effects against pentylenetetrazolinduced seizures through promoting the antioxidant, antifibrotic, anti-inflammatory, antiapoptotic pathways and inhibiting autophagy.
... The anti-inflammatory action of CRY is probably due to its ability to inhibit the interaction of cytokines with their target cells (Oliveira-Tintino et al. 2018). In addition, anticonvulsant (de Oliveira et al. 2016) and antimalarial activity of CRY have been reported (de Souza et al. 2017). The ability of CRYO to synergistically restore the sensitivity of tumor cells to doxorubicin in vitro has been shown (Di Giacomo et al. 2017). ...
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Several ATP-binding cassette (ABC) proteins reduce intracellular concentrations of antitumor drugs and hence weaken the response of cancer cells to chemotherapy. Accordingly, the inhibition of these export pumps constitutes a promising strategy to chemosensitize highly chemoresistant tumors, such as hepatocellular carcinoma (HCC). Here, we have investigated the ability of β-caryophyllene oxide (CRYO), a naturally occurring sesquiterpene component of many essential oils, to inhibit, at non-toxic doses, ABC pumps and improve the response of HCC cells to sorafenib. First, we have obtained a clonal subline (Alexander/R) derived from human hepatoma cells with enhanced multidrug resistance (MDR) associated to up-regulation (mRNA and protein) of MRP1 and MRP2. Analysis of fluorescent substrates export (flow cytometry) revealed that CRYO did not affect the efflux of fluorescein (MRP3, MRP4 and MRP5) but inhibited that of rhodamine 123 (MDR1) and calcein (MRP1 and MRP2). This ability was higher for CRYO than for other sesquiterpenes assayed. CRYO also inhibited sorafenib efflux, increased its intracellular accumulation (HPLC–MS/MS) and enhanced its cytotoxic response (MTT). For comparison, the effect of known ABC pumps inhibitors was also determined. They induced strong (diclofenac on MRPs), modest (verapamil on MDR1) or null (fumitremorgin C on BCRP) effect on sorafenib efflux and cytotoxicity. In the mouse xenograft model, the response to sorafenib treatment of subcutaneous tumors generated by mouse hepatoma Hepa 1–6/R cells, with marked MDR phenotype, was significantly enhanced by CRYO co-administration. In conclusion, at non-toxic dose, CRYO is able to chemosensitizating liver cancer cells to sorafenib by favoring its intracellular accumulation.
... BCP, orally administered at the dosages of 10, 30, and 100 mg/kg in C57BL/6 mice, exerted a dose-dependent anticonvulsant effect [9,43]. Furher research on mice has revealed that the sesquiterpene inhibits tonic-clonic seizures in MES (maximal electro shock seizure) test and reduces kainate-induced neurotoxicity, suggesting a possible use of the sesquiterpene for the treatment of epilepsy [44]. ...
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β-Caryophyllene (BCP), a natural bicyclic sesquiterpene, is a selective phytocannabinoid agonist of type 2 receptors (CB2-R). It isn't psychogenic due to the absence of an affinity to cannabinoid receptor type 1 (CB1). Among the various biological activities, BCP exerts anti-inflammatory action via inhibiting the main inflammatory mediators, such as inducible nitric oxide synthase (iNOS), Interleukin 1 beta (IL-1β), Interleukin-6 (IL-6), tumor necrosis factor-alfa (TNF-α), nuclear factor kapp a-light-chain-enhancer of activated B cells (NF-κB), cyclooxygenase 1 (COX-1), cyclooxygenase 2 (COX-2). Peroxisome proliferator-activated receptors alpha (PPAR-α) effects are also mediated by the activation of PPAR-α and PPAR-γ receptors. In detail, many studies, in vitro and in vivo, suggest that the treatment with -caryophyllene improves the phenotype of animals used to model various inflammatory pathologies, such as nervous system diseases (Parkinson's disease, Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, stroke), atherosclerosis, and tumours (colon, breast, pancreas, lymphoma, melanoma and glioma cancer). Furthermore, pre-clinical data have highlighted that BCP is potentially useful in Streptococcus infections, osteoporosis, steatohepatitis, and exerts anticonvulsant, analgesic, myorelaxing, sedative, and antidepressive effects. BCP is non-toxic in rodents, with a Lethal dose, 50% (LD50) greater than 5000 mg/kg. Nevertheless, it inhibits various cytochrome P450 isoforms (above all, CYP3A4), which metabolise xenobiotics, leading to adverse effects, due to drug levels over therapeutic window. All the reported data have highlighted that both pharmacological and toxicological aspects need to be further investigated with clinical trials.
... Another possible explanation is that these chemovars differ in concentrations of other phytocannabinoids as well (such as cannabidivarin), some of which have demonstrable antiepileptic effects in other models (Perucca, 2017;Rosenberg et al., 2017). Indeed, review of the supplier's material suggests that these chemovars differ in their terpene profiles, some of which may have therapeutic effects (de Oliveira et al., 2016;Russo, 2011;Nuutinen, 2018; see website links in Section 2). Clearly, any effects or proposed mechanisms must leave room for the possibility of yet unknown interactions or components of cannabis chemovars. ...
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There is significant interest in the use of cannabinoids for treatment of many epilepsies including absence epilepsy (AE). Genetic Absence Epilepsy Rats from Strasbourg (GAERS) model many aspects of AE including the presence of spike‐and‐wave discharges (SWDs) on electroencephalogram (EEG) and behavioural comorbidities, such as elevated anxiety. However, the effects of cannabis plant‐based phytocannabinoids have not been tested in GAERS. Therefore, we investigated how SWDs in GAERS are altered by the 2 most common phytocannabinoids, Δ⁹‐tetrahydrocannabinol (THC) and cannabidiol (CBD), and exposure to smoke from 2 different chemovars of cannabis. Animals were implanted with bipolar electrodes in somatosensory cortex and EEGs recorded for 2 hours. Injected THC (1‐10 mg/kg, i.p.) dose‐dependently increased SWDs to over 200% of baseline. In contrast, CBD (30‐100 mg/kg, i.p.) produced a ~50% reduction in SWDs. Exposure to smoke from a commercially available chemovar of high‐THC cannabis (Mohawk, Aphria Inc.) increased SWDs whereas a low‐THC/high‐CBD chemovar of cannabis (Treasure Island, Aphria Inc.) did not significantly affect SWDs in GAERS. Pre‐treatment with a CB1R antagonist (SR141716A) did not prevent the high‐THC cannabis smoke from increasing SWDs suggesting that the THC‐mediated increase may not be CB1R‐dependent. Plasma concentrations of THC and CBD were similar to previously reported values following injection and smoke exposure. Compared to injected CBD, it appears Treasure Island did not increase plasma levels sufficiently to observe an anti‐epileptic effect. Together these experiments provide initial evidence that acute phytocannabinoid administration exerts biphasic modulation of SWDs and may differentially impact patients with AE.
... Several studies revealed that stress in the prenatal period increases TBARS, which is an end-product of lipid peroxidation [28][29][30]. Besides that, seizure activity also increases lipid peroxidation [31,32]. Similarly, this study demonstrated that PS increases TBARS and MPO levels in PTZ group significantly. ...
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Objectives: This study aimed to investigate the effects of prenatal stress (PS) on hippocampus of early acute pentylenetetrazole (PTZ)-kindled offspring in adolescence. Recognition memory, morphological changes and synaptophysin levels in hippocampus were evaluated. Methods: Restraint stress was induced to a group of pregnant dams and non-stressed (NA) group remained undisturbed. Next, male and female offspring were divided as 1. PS-PTZ, 2. PS-control, 3. NA-PTZ and 4. NA-control (n = 12 in each group). The object recognition test was performed following PTZ injection (45 mg/kg) on postnatal day 10 (P10). Brains were collected on postnatal day 35 (P35) to determine neuronal density and synaptophysin expression by immuno/-histological studies. Further, oxidative stress products in hippocampus were analyzed with different biochemical assays. Results: PS impaired recognition memory in PTZ group significantly (p = 0.03); however, the impairment of PS was reversible in control group compared to PTZ (p = 0.04). Furthermore, PS caused neuronal loss in CA1 (p = 0.01) and decreased synaptophysin expression in the CA3 area of hippocampus in PTZ group (p = 0.03). PS also increased the oxidative stress markers in PTZ group significantly (p < 0.05). Conclusions: These results suggest that PS causes neurodevelopmental deficits in adolescent hippocampus and recognition memory after early-life seizures prominently. However, the damage of only PS in adolescence can be reversible. Therefore, the effects of PS in the adult hippocampus and other regions of brain need to be further studied
... BCP is approved by the FDA as a food additive, and is considered as a "generally recognized as safe" food or cosmetic addition [2]. BCP, a bicyclic sesquiterpene, displayed numerous pharmacological actions, including antioxidant [3], antiinflammatory [4], anticancer, analgesic [5], anticonvulsant [6], lipid lowering [7], and antispasmodic [8] effects. Furthermore, BCP demonstrated antioxidant, anti-inflammatory, and re-epithelialization activities in a rat skin wound excision model [9], an antidepressant-like effect in restraint plus stress-induced depression [10], improved the systemic inflammation and oxidative status of arthritic rats [11], and protected rat from carbon tetrachlorideinduced hepatic fibrosis [3]. ...
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Background: β-caryophyllene (BCP), a natural sesquiterpene, is extensively present in the essential oils of several plants. Cyclophosphamide (CYC) is an anticancer drug. However, its clinical usage is inadequate due to its cardiotoxicity. The aim of this study was to study the effects of BCP on cardiac injury induced by CYC exposure, and to identify the underlying mechanism of action. Methods: Five groups of Wistar rats were allocated. Group I (Normal), II (BCP), and III (CYC) acted as controls. Group IV, V (CYC + BCP) received BCP in two doses (100 and 200 mg/kg, orally, respectively) for 14 days after CYC challenge. CYC groups received 200 mg/kg, i.p. of the drug once on the first day of experiments. Results: CYC group displayed numerous ECG and histological irregularities and cardiac markers elevation. CYC induced lipid peroxidation and oxidative stress intensification, as well as inflammatory and apoptotic markers escalation. Treatment with BCP resulted in modified ECG traces and histological sections. BCP mitigated cardiac markers and lipid peroxidation whereas intensified antioxidant capacity. BCP activated Nrf2, with subsequent HO1 and NQO1 amplification. BCP diminished TLR4/NFκB pathway, which consequently lessened the inflammatory and apoptosis responses. Conclusion: BCP administration was associated with activated Nrf2/HO1/NQO1 and inhibited TLR4/NFκB pathways with subsequent enhanced anti-oxidative capacity and diminished inflammatory and apoptosis responses.
... However, anxyolitic, anti-inflammatory, analgesic and other pharmacological properties were showed in animal studies [27]. Beta-caryophyllene is an agonist of cannabinoid receptor 2 (CBD2) and shows anti-inflammatory and anticonvulsant properties in experimental models [28][29][30]. In our study the higher concentration of beta-caryophyllene in hemp cannabis flowers highlight its potential in epilepsy management and pain control. ...
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Cannabis cultivation for medical purposes in Brazil has been increased in the last years. While cannabis crops are prohibited, hundreds patients have been granted with judicial authorizations and there is little information about the cultivation conditions, yields and chemical profiles of the plants. Cannabis plants contain hundreds of compounds, with cannabinoids and terpenes the main drivers of their toxicological and pharmacological properties. Besides the cannabinoids, terpene contents are useful for the chemotaxonomic classification of different varieties, and their role in forensic analyses should be further delineated. The present study monitored cannabis crops of fifteen participants who were granted special licenses by the Brazilian Courts in Rio de Janeiro and São Paulo. The cultivation conditions were monitored and five cannabinoids (tetrahydrocannabinol acid-THCA, tetrahydrocannabinol-THC, cannabidiolic acid-CBDA, cannabidiol-CBD and cannabinol-CBN) and nineteen terpenes were quantified in cannabis flowers. The total grow cycle of thirty-five cannabis plants ranged from10 to 24 weeks. The dry flower yields ranged 22 to 90 g per plant. Most cannabis specimens were CBD-rich varieties (CBD levels from 1.6 to 16.7%, and THC levels from 0.0 to 2.6%, n=22) used to treat epileptic patients. The THC-rich varieties contained CBD levels ranging from 0.03 to 0.8%, and THC levels from 0.7 to 20.1%, n=11. Fewer of the samples contained THC:CBD ratios of approximately 1:1 (CBD levels of 3.3 to 3.8% and THC levels of 2.2 to 3.7%, n=2). The most abundant terpenes in the cannabis flowers were beta-caryophyllene, alpha-humulene, guaiol and alpha-bisabolol. CBD-rich varieties showed significant higher levels of beta-caryophyllene and alpha-humulene in comparison with THC-rich varieties. Overall, the study herein provides data concerning medical cannabis crops grown in a region of Brazil that not only guide individual medical cannabis cultivation methods but also aid forensic analyses.
... (E)-caryophyllene, on the other hand, has been reported to exhibit antiprotozoal activity against the parasite Leishmania amazonensis, which causes leishmaniasis [31]. Additionally, sesquiterpenes have anticonvulsant [32], antifungal [33], and anti-inflammatory properties [34]. ...
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Chemical Composition and Antioxidant Activity of Essential Oils from Eugenia patrisii Essential oils (EOs) were extracted from Eugenia patrisii, E. punicifolia, and Myrcia tomentosa, specimens A and B, using hydrodistillation. Gas chromatography coupled with mass spectrometry (GC/MS) was used to identify the volatile constituents present, and the antioxidant capacity of EOs was determined using diphenylpicryl-hydrazyl (DPPH) and trolox equivalent antioxidant capacity (TEAC) assays. For E. patrisii, germacrene D (20.03%), bicyclogermacrene (11.82%), and (E)-caryophyllene (11.04%) were identified as the major constituents of the EOs extracted from specimen A, whereas specimen B primarily comprised γ-elemene (25.89%), germacrene B (8.11%), and (E)-caryophyllene (10.76%). The EOs of E. punicifolia specimen A contained β-Elemene (25.12%), (E)-caryophyllene (13.11%), and bicyclogermacrene (9.88%), while specimen B was composed of (E)-caryophyllene (11.47%), bicyclogermacrene (5.86%), β-pinene (5.86%), and γ-muurolene (5.55%). The specimen A of M. tomentosa was characterized by γ-elemene (12.52%), germacrene D (11.45%), and (E)-caryophyllene (10.22%), while specimen B contained spathulenol (40.70%), α-zingiberene (9.58%), and γ-elemene (6.89%). Additionally, the chemical composition of the EOs was qualitatively and quantitatively affected by the collection period. Furthermore, the EOs of the studied specimens, especially specimen A of E. punicifolia, showed a greater antioxidant activity in DPPH rather than TEAC, as represented by a significantly high inhibition percentage (408.0%).
... It was further shown that the CBdependent attenuation of M biosynthesis and AANAT activity is mediated by an intracellular interaction between CBs and the activated AANAT enzyme (Koch et al., 2006). Development of epileptic seizures and brain tissue lipid peroxidation are ameliorated by antioxidants like vitamin C (Ayyildiz et al., 2007), resveratrol (Mishra et al., 2015), β-caryophyllene (de Oliveira et al., 2016), and quercetin (Sefil et al., 2014). Cognition impairment in high-fat diet (HFD)-fed aged rats was alleviated by through antagonism of brain insulin resistance (Xu et al., 2019). ...
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Background Lipid profile and redox status play a role in brain (dys)functions. Cannabinoid and melatonergic systems operate in the brain and contribute to brain (patho)physiology, but their roles in the modulation of brain lipid and redox status are not well-known. We studied the effect of ethanol extract of Cannabis sativa (CS) and/or melatonin (M) on the lipid profile and anti-oxidant system of the rat brain. Methods We randomly divided twenty-four (24) female Wistar rats into 4 groups ( n = 6 rats each). Group 1 (control) received distilled water mixed with DMSO. Groups II–IV received CS (2 mg/kg), M (4 mg/kg), and co-administration of CS and M (CS + M) respectively via oral gavage between 8:00 am and 10:00 am once daily for 14 days. Animals underwent 12-h fasting after the last day of treatment and sacrificed under ketamine anesthesia (20 mg/kg; i.m). The brain tissues were excised and homogenized for assay of the concentrations of the total cholesterol (TC), triacylglycerol (TG), high-density lipoprotein cholesterol (HDL-C), nitric oxide (NO), malondialdehyde (MDA), and the activities of glucose-6-phosphate dehydrogenase (G6PD), glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), and acetylcholinesterase (AChE). One-way analysis of variance (ANOVA) was used to compare means across groups, followed by the least significant difference (LSD) post-hoc test. Results CS and/or M did not affect the lipid profile parameters. However, CS increased the G6PD (from 15.58 ± 1.09 to 21.02 ± 1.45 U/L; p = 0.047), GPx (from 10.47 ± 0.86 to 17.71 ± 1.04 U/L; p = 0.019), and SOD (from 0.81 ± 0.02 to 0.90 ± 0.01 μM; p = 0.007), but decreased NO (from 9.40 ± 0.51 to 6.75 ± 0.21 μM; p = 0.010) and had no effect on MDA ( p = 0.905), CAT ( p = 0.831), GR ( p = 0.639), and AChE ( p = 0.571) in comparison with the control group. M augmented the increase in G6PD (from 21.02 ± 1.45 U/L to 27.18 ± 1.81 U/L; p = 0.032) and decrease in NO (from 6.75 ± 0.21 to 4.86 ± 0.13 μM; p = 0.034) but abolished the increase in GPx (from 17.71 ± 1.04 to 8.59 ± 2.06 U/L; p = 0.006) and SOD (from 0.90 ± 0.01 to 0.70 ± 0.00 μM; p = 0.000) elicited by CS in the rat brain in comparison with the CS group. Conclusions CS and M do not alter brain lipid profile. Our data support the contention that CS elicits an anti-oxidative effect on the brain tissue and that CS + M elicits a pro-oxidant effect in rat brain.
... Extract or its vehicle was provided 60 minutes prior to the start of the test session. [10] Forced swim test was conducted based on the method previously described with some modifications [11] . Rats were individually placed in open cylindrical container (40 cm in diameter × 80 cm in height) containing water at 45 cm height. ...
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Introduction: To investigate the preventive effect of Phaseolus vulgaris seed coats on pentylenetetrazole-induced kindling and behavioral comorbidities. Material and Methods: Thirty Wistar albino rats were categorized into five groups. The first group received regular saline (0.9 % w/v NaCl) p.o.; the second group received PTZ (35 mg/kg b.w.) i.p.; the third group received valproic acid (200 mg/kg b.w.) p.o.; the fourth group received P. vulgaris extract (100 mg/kg b.w.) p.o.; the fifth group received P. vulgaris extract (200 mg/kg b.w.) p.o. on an alternate day for 21 days. PTZ improved lipid peroxidase levels, decreased Glutathione level, decreased superoxide dismutase activity, increased Nitric Oxide level. Result: This study revealed that P. vulgaris (Hydroalcoholic extract) increased the anti-oxidant level of both 100 mg/kg and 200 mg/kg compared to the PTZ category. Histopathological findings revealed that the hippocampal section of the brain of rats receiving P. vulgaris extract had improved relative to the receiving PTZ group. Conclusion: Based on the result, it is proposed that Phaseolus vulgaris has anti-oxidant properties. This is useful for the treatment of epilepsy.
... for 30 days) significantly decreased cauda epididymal sperm count, percentages of both motile, and with normal morphology sperm. Nevertheless, it neither affected the ultrastructure of the cauda epididymis or testes nor the histology of the treated rats (Al-Alami et al. 2015) as shown in Table 3. Due to its safe contraceptive dose for both male sexual and other organs and its selectivity towards antisperm action (post-testicular targeting), in addition to its reversibility (Correia et al. 2016;Tambe et al. 1996;Al-Alami et al. 2015), it may be considered as a promising natural male contraceptive (see Tables 3, 4, 5). Furthermore, it Phytochem Rev should be evaluated for 60 days instead of 30 days (complete spermatogenesis cycle of rats is from 48 to 60 days) by oral administration after enhancement of its poor oral bioavailability to accurately judge its safety and efficacy as shown in Table 5 (Fidyt et al. 2016;Di Sotto et al. 2018;Zeng et al. 2006). ...
... (E)-caryophyllene, on the other hand, has been reported to exhibit antiprotozoal activity against the parasite Leishmania amazonensis, which causes leishmaniasis [31]. Additionally, sesquiterpenes have anticonvulsant [32], antifungal [33], and anti-inflammatory properties [34]. ...
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Essential oils (EOs) were extracted from Eugenia patrisii, E. punicifolia, and Myrcia tomentosa, specimens A and B, using hydrodistillation. Gas chromatography coupled with mass spectrometry (GC/MS) was used to identify the volatile constituents present, and the antioxidant capacity of EOs was determined using diphenylpicryl-hydrazyl (DPPH) and trolox equivalent antioxidant capacity (TEAC) assays. For E. patrisii, germacrene D (20.03%), bicyclogermacrene (11.82%), and (E)-caryophyllene (11.04%) were identified as the major constituents of the EOs extracted from specimen A, whereas specimen B primarily comprised γ-elemene (25.89%), germacrene B (8.11%), and (E)-caryophyllene (10.76%). The EOs of E. punicifolia specimen A contained β-Elemene (25.12%), (E)-caryophyllene (13.11%), and bicyclogermacrene (9.88%), while specimen B was composed of (E)-caryophyllene (11.47%), bicyclogermacrene (5.86%), β-pinene (5.86%), and γ-muurolene (5.55%). The specimen A of M. tomentosa was characterized by γ-elemene (12.52%), germacrene D (11.45%), and (E)-caryophyllene (10.22%), while specimen B contained spathulenol (40.70%), α-zingiberene (9.58%), and γ-elemene (6.89%). Additionally, the chemical composition of the EOs was qualitatively and quantitatively affected by the collection period. Furthermore, the EOs of the studied specimens, especially specimen A of E. punicifolia, showed a greater antioxidant activity in DPPH rather than TEAC, as represented by a significantly high inhibition percentage (408.0%).
... (E)-caryophyllene, on the other hand, has been reported to exhibit antiprotozoal activity against the parasite Leishmania amazonensis, which causes leishmaniasis [31]. Additionally, sesquiterpenes have anticonvulsant [32], antifungal [33], and anti-inflammatory properties [34]. ...
... (E)-caryophyllene, on the other hand, has been reported to exhibit antiprotozoal activity against the parasite Leishmania amazonensis, which causes leishmaniasis [31]. Additionally, sesquiterpenes have anticonvulsant [32], antifungal [33], and anti-inflammatory properties [34]. ...
Article
Full-text available
Essential oils (EOs) were extracted from Eugenia patrisii, E. punicifolia, and Myrcia tomentosa, specimens A and B, using hydrodistillation. Gas chromatography coupled with mass spectrometry (GC/MS) was used to identify the volatile constituents present, and the antioxidant capacity of EOs was determined using diphenylpicryl-hydrazyl (DPPH) and trolox equivalent antioxidant capacity (TEAC) assays. For E. patrisii, germacrene D (20.03%), bicyclogermacrene (11.82%), and (E)-caryophyllene (11.04%) were identified as the major constituents of the EOs extracted from specimen A, whereas specimen B primarily comprised γ-elemene (25.89%), germacrene B (8.11%), and (E)-caryophyllene (10.76%). The EOs of E. punicifolia specimen A contained β-Elemene (25.12%), (E)-caryophyllene (13.11%), and bicyclogermacrene (9.88%), while specimen B was composed of (E)-caryophyllene (11.47%), bicyclogermacrene (5.86%), β-pinene (5.86%), and γ-muurolene (5.55%). The specimen A of M. tomentosa was characterized by γ-elemene (12.52%), germacrene D (11.45%), and (E)-caryophyllene (10.22%), while specimen B contained spathulenol (40.70%), α-zingiberene (9.58%), and γ-elemene (6.89%). Additionally, the chemical composition of the EOs was qualitatively and quantitatively affected by the collection period. Furthermore, the EOs of the studied specimens, especially specimen A of E. punicifolia, showed a greater antioxidant activity in DPPH rather than TEAC, as represented by a significantly high inhibition percentage (408.0%).
... In addition, it has been seen that the monoterpenes β-caryophyllen (De Oliveira et al., 2016), β-pinene and α-pinene (Felipe et al., 2019) present in EOPT may also be involved in the observed effect with involving of GABAergic system. According to Guzmán-Gutiérrez et al. (2012), when these compounds were administered in mice, there is a reduction of CN in the open field test. ...
Article
This study aims to investigate the toxicity and possible effects and mechanism of action of the essential oil obtained from the Piper tuberculatum Jacq.‘s fruit (EOPT) on the central nervous system (CNS) of mice. In the Hippocratic test, it was observed ambulation reduction, sedation, piloerection and mortality of 25% at the dose of 1000 mg/kg and 50% at the dose of 5000 mg/kg. The results found demonstrate that the EOPT is endowed with a possible depressant effect, and is characterized by a likely sedative (open field test), anxiolytic-like (elevated plus maze test), anticonvulsant (pentylenetetrazole-induced seizure test) and hypnotic action (pentobarbital-induced sleeping time). These effects may possibly occur due to the interaction between the monoterpenes, present in its essential oil, and the GABAergic pathway confirmed by reversion of the effects by flumazenil. Then, these results demonstrate the therapeutic potential and the validation of the ethnopharmacological use of this species in the treatment of disorders affecting the central nervous system.
... Its biological activities are widely reported in the literature and include antimicrobial [14], antiproliferative, and antiprotozoal [38]. There are also reports of its anticonvulsant [39], analgesic, and anti-inflammatory properties [40]. ...
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The essential oil (EO) of Calycolpus goetheanus (Myrtaceae) specimens (A, B, and C) were obtained through hydrodistillation. The analysis of the chemical composition of the EOs was by gas chromatography coupled with mass spectrometry CG-MS, and gas chromatography coupled with a flame ionization detector CG-FID. The phytotoxic activity of those EOs was evaluated against two weed species from common pasture areas in the Amazon region: Mimosa pudica L. and Senna obtusifolia (L.) The antioxidant capacity of the EOs was determined by (DPPH•) and (ABTS•+). Using molecular docking, we evaluated the interaction mode of the major EO compounds with the molecular binding protein 4-hydroxyphenylpyruvate dioxygenase (HPPD). The EO of specimen A was characterized by β-eudesmol (22.83%), (E)-caryophyllene (14.61%), and γ-eudesmol (13.87%), while compounds 1,8-cineole (8.64%), (E)-caryophyllene (5.86%), δ-cadinene (5.78%), and palustrol (4.97%) characterize the chemical profile of specimen B’s EOs, and specimen C had α-cadinol (9.03%), δ-cadinene (8.01%), and (E)-caryophyllene (6.74%) as the majority. The phytotoxic potential of the EOs was observed in the receptor species M. pudica with percentages of inhibition of 30%, and 33.33% for specimens B and C, respectively. The EOs’ antioxidant in DPPH• was 0.79 ± 0.08 and 0.83 ± 0.02 mM for specimens A and B, respectively. In the TEAC, was 0.07 ± 0.02 mM for specimen A and 0.12 ± 0.06 mM for specimen B. In the results of the in silico study, we observed that the van der Waals and hydrophobic interactions of the alkyl and pi-alkyl types were the main interactions responsible for the formation of the receptor–ligand complex.
... A similar entourage effect between cannabinoids and terpenes was previously described for mood disorder, as well as for pain [12,13]. Furthermore, some representative terpenes of K2, such as β-caryophyllene (a CB 2 receptors agonist), β-myrcene and limonene have displayed anticonvulsant activity against seizures induced by pentylenetetrazol [31,32]. Moreover, linalool has been able to act as an anticonvulsant in quinolinic acid-induced seizures in vivo [33] and to alleviate maximal electroshock-induced seizures in mice [34]. ...
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Compounds present in Cannabis sativa L. preparations have recently attracted much attention in the treatment of drug-resistant epilepsy. Here, we screened two olive oil extracts from a non-psychoactive C. sativa variety, fully characterized by high-performance liquid chromatography and gas chromatography. Particularly, hemp oils with different concentrations of terpenes were administered at the same dose of cannabidiol (25 mg/kg/day orally), 1 h before the 6-Hz corneal stimulation test (44 mA). Mice were stimulated once a day for 5 days and evaluated by videoelectrocorticographic recordings and behavioral analysis. Neuronal activation was assessed by FosB/deltaFosB immunoreactivity. Both oils significantly reduced the percentage of mice experiencing convulsive seizures in comparison to olive oil-treated mice (p < 0.050; Fisher’s exact test), but only the oil enriched with terpenes (K2) significantly accelerated full recovery from the seizure. These effects occurred in the presence of reduced power of delta rhythm, and, instead, increased power of theta rhythm, along with a lower FosB/DFosB expression in the subiculum (p < 0.050; Duncan’s method). The overall findings suggest that both cannabinoids and terpenes in oil extracts should be considered as potential therapeutic agents against epileptic seizures and epilepsy.
Article
This review focuses on the possible roles of phytocannabinoids, synthetic cannabinoids, endocannabinoids, and “transient receptor potential cation channel, subfamily V, member 1” (TRPV1) channel blockers in epilepsy treatment. The phytocannabinoids are compounds produced by the herb Cannabis sativa, from which Δ⁹-tetrahydrocannabinol (Δ⁹-THC) is the main active compound. The therapeutic applications of Δ⁹-THC are limited, whereas cannabidiol (CBD), another phytocannabinoid, induces antiepileptic effects in experimental animals and in patients with refractory epilepsies. Synthetic CB1 agonists induce mixed effects, which hamper their therapeutic applications. A more promising strategy focuses on compounds that increase the brain levels of anandamide, an endocannabinoid produced on-demand to counteract hyperexcitability. Thus, anandamide hydrolysis inhibitors might represent a future class of antiepileptic drugs. Finally, compounds that block the TRPV1 (“vanilloid”) channel, a possible anandamide target in the brain, have also been investigated. In conclusion, the therapeutic use of phytocannabinoids (CBD) is already in practice, although its mechanisms of action remain unclear. Endocannabinoid and TRPV1 mechanisms warrant further basic studies to support their potential clinical applications. This article is part of the Special Issue “NEWroscience 2018".
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Introduction Phlomoides species have long been used in ethnomedicine for various purposes, such as stomach disorders and wound healing. The aim of this study was to investigate some phytochemical and biological features of one species, Phlomoides labiosa. Methods The aerial part, rhizome, and the seed of P. labiosa were extracted using various solvents with increasing polarity. Essential oil and fatty acid contents were analyzed via GC/MS method. Antioxidant, antibacterial, and cytotoxic properties were evaluated in all samples. After that, extract nanoliposomes were created to improve the biological effects by increasing the solubility. Results Obtained results indicated that oleic acid is assumed as the major component of seed oil with a total fatty acid content of 37.62 (w/w %). Ethyl acetate and methanol extracts indicated the antioxidant activity in line with the phenol contents. The most potent antibacterial effects were associated with the ethyl acetate extracts of aerial part and seed. Besides, the dichloromethane extract of the aerial part showed the highest cytotoxicity among all tested samples (IC50=40.88±16.05 µg/mL). Despite the hypothesis of the better effect of encapsulated extracts, nanoparticles indicated lower biological activities than free forms. Conclusions Because of the high amount of oleic acid and overall yield, the seed oil may be suitable for nutritional reasons. The antibacterial activity of ethyl acetate extract and the considerable cytotoxicity of dichloromethane extract make them potential candidates for future assessments. Limited extract release from the membranes of nanoliposomes may be the main reason for biological activity reduction.
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Neuroprotective agents are able to defend the central nervous system against acute or chronic neuronal injuries. Even with the progress made over the last decades, most of the medications prescribed for the management of neurodegenerative diseases can only reduce their symptoms and slow down their progression. Based on natural product research, there are potential effective medicinal plants and phytochemicals for modulating neuronal functions and protecting against neurodegeneration. Plants in the genus Pistacia are also among valuable natural resources for neuroprotection research based on experiences in traditional medicine. Studies have supported the value of bioactive compounds of the genus Pistacia for central nervous system disorders such as Alzheimerʼs, Parkinsonʼs, multiple sclerosis, cerebral ischemia, depression, and anxiety. Related literature has also revealed that most of the evidence on neuroprotection in the genus Pistacia is in the form of preliminary studies, mainly including models of behavior, motor function, and memory impairments in animals, neural toxicity, cerebral ischemia and seizure models, evaluation of their effects on antioxidant and inflammatory biomarkers, amyloid β aggregation, and acetylcholinesterase as well as investigations into some cellular pathways. Along with the phytonutrients in kernels such as pistachios, various phytochemicals, mostly terpenes, and phenolic compounds have also been identified in different plant parts, in particular their oleoresins, of species in the genus Pistacia. In this review, the pharmacology of neurological effects and related molecular mechanisms of the plants belonging to the genus Pistacia and its active constituents, as well as pharmacokinetics aspects, are discussed.
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Background/aims: Helicobacter pylori (H. pylori) infections, which cause a variety of gastrointestinal symptoms, are common in South Korea. Recent reports have shown a decline in the H. pylori eradication rates. β-caryophyllene is a natural bicyclic sesquiterpene that occurs in a wide range of plant species, such as cloves, basil, and cinnamon. β-caryophyllene has been reported to have anti-inflammatory and anti-bacterial effects. This study investigated the inhibitory effects of β-caryophyllene on H. pylori and its potential role as an alternative gastrointestinal drug. Methods: This 8-week, randomized double-blind, placebo-controlled trial categorized subjects into a β-caryophyllene group (33 patients who received 126 mg/day of β-caryophyllene) and a placebo group (33 patients who received a placebo preparation). The inflammation level of H. pylori infiltration and the eradication rates were evaluated endoscopically and with the urea breath test (UBT) in both groups before and after administering the medication. The serum cytokine levels (tumor necrosis factor-α, and interleukin [IL]-1β and IL-6) were compared in both groups before and after administering the medication. Results: Complete eradication was not observed in either group. Moreover, there was no significant change in the UBT and updated Sydney score. On the other hand, the β-caryophyllene group showed significant improvement in nausea (p=0.025) and epigastric pain (p=0.018), as well as a decrease in the serum IL-1β levels (p=0.038). Conclusions: β-caryophyllene improves dyspepsia symptoms and can be considered a useful supplementary treatment for gastrointestinal disease.
Article
Objective: The endocannabinoid system (ECS) is comprised of cannabinoid receptors 1 and 2 (CB1R and CB2R), endogenous ligands, and regulatory enzymes, and serves to regulate several important physiological functions throughout the brain and body. Recent evidence suggests that the ECS may be a promising target for the treatment of epilepsy, including epilepsy subtypes that arise from mutations in the voltage-gated sodium channel SCN1A. The objective of this study was to explore the effects of modulating CB2R activity on seizure susceptibility. Methods: We examined susceptibility to induced seizures using a number of paradigms in CB2R knockout mice (Cnr2-/- ), and determined the effects of the CB2R agonist, JWH-133, and the CB2R antagonist, SR144528, on seizure susceptibility in wild-type mice. We also examined seizure susceptibility in Cnr2 mutants harboring the human SCN1A R1648H (RH) epilepsy mutation and performed Electroencephalography (EEG) analysis to determine whether the loss of CB2Rs would increase spontaneous seizure frequency in Scn1a RH mutant mice. Results: Both heterozygous (Cnr2+/- ) and homozygous (Cnr2-/- ) knockout mice exhibited increased susceptibility to pentylenetetrazole (PTZ)-induced seizures. The CB2R agonist JWH-133 did not significantly alter seizure susceptibility in wild-type mice; however, administration of the CB2R antagonist SR144528 resulted in increased susceptibility to PTZ-induced seizures. In offspring from a cross between the Cnr2 × RH lines, both Cnr2 and RH mutants were susceptible to PTZ-induced seizures; however, seizure susceptibility was not significantly increased in mutants expressing both mutations. No spontaneous seizures were observed in either RH or Cnr2/RH mutants during 336-504 hours of continuous EEG recordings. Significance: Our results demonstrate that reduced CB2R activity is associated with increased seizure susceptibility. CB2Rs might therefore provide a therapeutic target for the treatment of some forms of epilepsy.
Article
Background Central nervous system disorders such as anxiety, depression and epilepsy are characterized by sharing several molecular mechanisms in common and the involvement of the L-arginine/NO pathway in neurobehavioral studies with β-caryophyllene is still little discussed. Objectives One of the objectives of the present study was to demonstrate the anxiolytic behavioral effect of β-caryophyllene (β-CBP) in female Swiss mice, as well as to investigate the molecular mechanisms underlying the results obtained. Methods This study evaluated the neurobehavioral effects of β-CBP using the open field test, rota-rod test, elevated plus maze test, novelty suppressed feeding test, tail suspension test and forced swim test, as well as pilocarpine, pentylenetetrazole and isoniazid-induced epileptic seizure models. Results: The results demonstrated that the neuropharmacological activities of β-CBP may involve benzodiazepine/GABAergic receptors, since the pre-treatment of β-CBP (200 mg/kg) associated with flumazenil (5 mg/kg, benzodiazepine receptor antagonist) and bicuculline (1 mg/kg, selective GABAA receptor antagonist) reestablished the anxiety parameters in the elevated plus-maze test, as well as the results of reduced latency to consume food in the novelty suppressed feeding test. In addition to benzodiazepine/GABAergic receptors, the neuropharmacological properties of β-CBP may be related to inhibition of nitric oxide synthesis, since pre-treatment with L-arginine (500- 750 mg/kg) reversed significantly the anxiolytic, antidepressant and anticonvulsant activities of β-CBP. Conclusion The results obtained provide additional support in understanding the neuromolecular mechanisms underlying the anxiolytic, antidepressant and anticonvulsive properties of β-CBP in female Swiss mice.
Article
Background Status epilepticus (SE) is a neurological life-threatening condition, resulting from the failure of the mechanisms responsible for seizure termination. SE is often pharmacoresistant and associated with significant morbidity and mortality. Hence, ceasing or attenuating SE and its consequences is of fundamental importance. Beta-caryophyllene is a functional CB2 receptor agonist and exhibit a good safety profile. Besides, it displays beneficial effects in several experimental conditions, including neuroprotective activity. In the present study we aimed to investigate the effects of beta-caryophyllene on pilocarpine-induced SE. Methods Wistar rats were submitted to pilocarpine-induced SE and monitored for 24 hours by video and EEG for short-term recurrence of seizure activity (i.e. seizures occurring within 24 hours after termination of SE). Rats received beta-caryophyllene (100 mg/kg, ip) at 1, 8- and 16-hours after SE. Twenty-four hours after SE we evaluated sensorimotor response, neuronal damage (fluoro jade C staining) and serum albumin infiltration into brain parenchyma. Results Beta-caryophyllene-treated animals presented fewer short-term recurrent seizures than vehicle-treated counterparts, suggesting an anticonvulsant effect after SE. Behavioral recovery from SE and the number of fluoro jade C positive cells in the hippocampus and thalamus were not modified by beta-caryophyllene. Treatment with beta-caryophyllene attenuated the SE-induced increase of albumin immunoreactivity in the hippocampus, indicating a protective effect against blood-brain-barrier breakdown. Conclusions Given the inherent difficulties in the treatment of SE and its consequences, present results suggest that beta-caryophyllene deserve further investigation as an adjuvant therapeutic strategy for SE.
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Epilepsy is a common chronic neurological disease. The hallmark of epilepsy is recurrent, unprovoked seizures. Unfortunately, drug resistance is frequent in patients with epilepsy, and therefore improved therapeutic strategies are needed. In the present study, we tested the effect of pregabalin in association with beta-caryophyllene, an FDA-approved food additive and naturally occurring agonist of cannabinoid receptor subtype 2 against pentylenetetrazol (PTZ)-induced seizures in rats. In addition, selected neurochemical parameters were evaluated in the cerebral cortex. Adult male Wistar rats received beta-caryophyllene (100 mg/kg), pregabalin (40 mg/kg) or their combination before PTZ (60 mg/kg). Appropriated vehicle-treated control groups were included for each treatment. Animals were monitored by video-EEG and the latency to myoclonic seizures, latency to tonic-clonic seizures, tonic-clonic seizure duration and overall seizure score were measured. Glial fibrillary acidic protein (GFAP) release, erythroid-related factor 2 (Nrf2), c-fos and 3-nytrotyrosine (3-NT) levels were evaluated in the frontal cortex. We found that beta-caryophyllene plus pregabalin increased the latency to PTZ-induced myoclonic and tonic-clonic seizures and decreased the tonic-clonic seizure duration and overall seizure score. Interestingly, lower levels of GFAP, c-Fos and 3-NT were observed in animals receiving beta-caryophyllene and pregabalin treatments. Our results suggest a possible synergic effect of beta-caryophyllene plus pregabalin against PTZ induced-seizures.
Article
β-Caryophyllene (BCP) is a bicyclic sesquiterpene compound that has anti-diabetic activity. However, the effect of BCP on diabetic nephropathy (DN) remains unclear. Here, we aimed to evaluate the potential role of BCP in high glucose (HG)-induced glomerular mesangial cells (MCs). MCs were maintained under HG condition to simulate DN in vitro. Our results showed that BCP inhibited HG-induced cell proliferation, ROS production and NADPH oxidase (NOX) 2/4 expression. BCP exhibited anti-inflammatory activity with decreased levels of TNF-α, IL-1β, IL-6 in HG-induced MCs. Moreover, BCP treatment suppressed the HG-induced secretion of fibronectin (FN) and collagen IV (Col IV) in MCs. Furthermore, BCP suppressed the NF-κB activation and enhanced the Nrf2 activation in HG-induced MCs. However, inhibition of Nrf2 attenuated the protective effects of BCP on HG-induced MCs, while inhibition of NF-κB enhanced the nephro-protective effects of BCP on MCs. In conclusion, these findings demonstrated that BCP executed protective effects on HG-induced MCs via regulating NF-κB and Nrf2 signaling pathways.
Article
Aflatoxin B1 (AFB1) is a mycotoxin highly toxic and carcinogenic to humans due to its potential to induce oxidative stress. The Beta-caryophyllene (BCP) have been highlighted for its broad spectrum of pharmacological effects. The present study aimed to investigate the beneficial effects of BCP against the susceptibility of hepatic and renal tissues to AFB1 toxicity, in biochemical parameters to assess organ function, tissue oxidation, and the immunocontent of oxidative and inflammatory proteins. Male Wistar rats was exposed to AFB1 (250 μg/kg, i.g.) and/or BCP (100 mg/kg, i.p.) for 14 successive days. It was found that exposure to AFB1 did not change the measured renal toxicity parameters. Also, AFB1 increased liver injury biomarkers (gamma glutamyl transferase and alkaline phosphatase) and reduced levels of non-enzymatic antioxidant defenses (ascorbic acid and non-protein thiol), however did not cause changes in the lipid peroxidation levels. Moreover, AFB1 interfered in oxidative pathway regulated by Kelch-like ECH-associated protein (Keap1)/nuclear factor (erythroid-derived 2)-like 2 (Nrf2), overacting Glutathione-S-Transferase (GST) activity. Lastly, a main effect of AFB1 on the total interleukin 1 beta (IL-1β) was observed. Remarkably, the associated treatment of AFB1 + BCP improved altered liver parameters. In addition, BCP and AFB1 + BCP groups showed an increase in the levels of inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ). Thus, these results indicated that BCP has potential protective effect against AFB1 induced hepatotoxicity.
Article
Ethnopharmacological relevance Epilepsy is one of the most commonly occurring non-communicable neurological disorder that affects people of all age groups. Around 50 million people globally are epileptic, with 80% cases in developing countries due to lack of access to treatments determined by high cost and poor availability or it can be defined by the fraction of active epileptic patients who are not appropriately being treated. The availability of antiepileptic drugs and their adjuvant therapy in such countries is less than 50% and these are highly susceptible to drug interactions and severe adverse effects. As a result, the use of herbal medicine is increasingly becoming popular. Aim of the study To provide pharmacological information on the active constituents evaluated in the preclinical study to treat epilepsy with potential to be used as an alternative therapeutic option in future. It also provides affirmation for the development of novel antiepileptic drugs derived from medicinal plants. Materials and methods Relevant information on the antiepileptic potential of phytoconstituents in the preclinical study (in-vitro, in-vivo) is provided based on their effect on screening parameters. Besides, relevant information on pharmacology of phytoconstituents, the traditional use of their medicinal plants related to epilepsy and status of phytoconstituents in the clinical study were derived from online databases, including PubMed, Clinicaltrial. gov, The Plant List (TPL, www.theplantlist.org), Science Direct. Articles identified using preset searching syntax and inclusion criteria are presented. Results More than 70% of the phytoconstituents reviewed in this paper justified the traditional use of their medicinal plant related to epilepsy by primarily acting on the GABAergic system. Amongst the phytoconstituents, only cannabidiol and tetrahydrocannabinol have been explored for clinical application in epilepsy. Conclusion The preclinical and clinical data of the phytoconstituents to treat epilepsy and its associated comorbidities provides evidence for the discovery and development of novel antiepileptic drugs from medicinal plants. In terms of efficacy and safety, further randomized and controlled clinical studies are required to understand the complete pharmacodynamic and pharmacokinetic picture of phytoconstituents. Also, specific botanical source evaluation is needed.
Article
Background Both rosin and turpentine are extremely abundant and renewable resources in nature. p-Cymene and (−)-β-caryophyllene are components of turpentine and dehydroabietic acid is a component of rosin. The determination of their vapor-liquid equilibrium data can provide more complete basic data for the separation and purification of rosin and turpentine systems. Methods The isothermal vapor-liquid equilibrium data of the systems of p-cymene (1) + (−)-β-caryophyllene (2) and p-cymene (1) + (−)-β-caryophyllene (2) + dehydroabietic acid (3) were determined by headspace gas chromatography at resin tapping temperature of 313.15, 323.15, and 333.15 K. Significant findings The experimental data showed that the addition of dehydroabietic acid significantly reduced the relative volatility of p-cymene to (−)-β-caryophyllene, and inhibited the separation of (−)-β-caryophyllene and p-cymene. The thermodynamic consistency of binary system was verified by Redlich Kister area test and Van Ness point test, while the thermodynamic consistency of ternary system was verified by Van Ness point test. The experimental data were compared with the calculated value by the NRTL, Wilson, and UNIQUAC models. The average absolute deviation of the vapor phase molar fraction was not more than 0.3%. In addition, the NRTL model was the most appropriate model.
Article
Mounting evidence suggests that modulation of cannabinoid 2 receptors (CB2Rs) is therapeutic in mouse models of neurological disorders, including neuropathic pain, neurodegenerative disease, and stroke. We previously showed that reducing CB2R activity increases seizure susceptibility in mice. In the present study, we evaluated the therapeutic potential of the CB2R positive allosteric modulator, Ec21a, against induced seizures in mice. The pharmacokinetic profile of Ec21 demonstrated a similar distribution in brain and plasma, with detection up to 12 hours following injection. Ec21a increased resistance to induced seizures in CF1 wild-type mice and mice harboring the SCN1A R1648H human epilepsy mutation. A rotarod test provided evidence that Ec21a does not cause neurotoxicity-induced motor deficits at its therapeutic dose, and seizure protection was maintained with repeated drug administration. The selectivity of Ec21a for CB2R was supported by the ability of the CB2R antagonist AM630, but not the CB1R antagonist AM251, to block Ec21a-conferred seizure protection in mice, and a lack of significant binding of Ec21a to 34 brain-expressed receptors and transporters in vitro. These results identify allosteric modulation of CB2Rs as a promising therapeutic approach for the treatment of epilepsy.
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Artemisia amygdalina, a critically endangered aromatic plant species, which is endemic to subalpine habitats of North Western Himalayas, serves as a rich repository of pharmacologically active constituents. Despite being medicinally imperative, this Artemisia species has acquired diminutive attention and is presently limited to miniature populations in Kashmir Himalayan region. Therapeutically active phytoconstituents of this plant species encompass alkaloids, cardiac glycosides, phenolics, steroids, tannins, terpenes, and other essential oils, with 1,8-cineole, 3-carene, artemisinin, artesunate, camphene, ludartin, p-cymene, santonin, α-cadinol, α-pinene, and β-pinene being the characteristic compounds isolated so far. Several therapeutic activities ascribed to these essential bioactive compounds extracted from this species include analgesic, antibacterial, anticancerous, anticonvulsant, antidepressant, antidiabetic, antifungal, antihelminthic, antiinflammatory, antimalarial, antioxidant, antispasmodic, antiulcer, antiviral, anxiolytic, sedative, and immunomodulatory properties. Elaborate bioassay of pharmacologically active phytoconstituents of this species is yet amenable for further exploration.
Article
This study aimed to investigate, through in vivo and in vitro methodologies, the effect of acute trans,trans-farnesol (12.5, 25, 50 or 100 mg/kg, p.o.) administration on behavioral and neurochemical parameters associated with pilocarpine-induced epileptic seizure (300 mg/kg, i.p.) in mice. The initial results showed that the compound in question presents no anxiolytic-like or myorelaxant effects, despite reducing locomotor activity in the animals at all doses tested. In addition, the lowest dose increased the latency to onset of the first epileptic seizure, and the time to death. In addition to decreasing the mortality percentage in mice submitted to the pilocarpine model. In this same model, pretreatment with the lowest dose of the compound decreased the hippocampal concentrations of thiobarbituric acid and nitrite, and partially restored striatal concentrations of noradrenaline, dopamine, and serotonin. Taken together, the results suggest that trans,trans-farnesol presents a central depressant effect which contributes to its antiepileptic action which, in turn, seems to be mediated by the antagonism of muscarinic cholinergic receptors, reduction of oxidative stress. and modulation of noradrenaline, dopamine and serotonin concentrations in the central nervous system.
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Despite the introduction of over 15 third-generation anti-epileptic drugs, current medications fail to control seizures in 20-30% of patients. However, our understanding of the mechanisms mediating the development of epilepsy and the causes of drug resistance has grown substantially over the past decade, providing opportunities for the discovery and development of more efficacious anti-epileptic and anti-epileptogenic drugs. In this Review we discuss how previous preclinical models and clinical trial designs may have hampered the discovery of better treatments. We propose that future anti-epileptic drug development may be improved through a new joint endeavour between academia and the industry, through the identification and application of tools for new target-driven approaches, and through comparative preclinical proof-of-concept studies and innovative clinical trials designs.
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Abstract β-Caryophyllene (trans-4,11,11-trimethyl-8-methylenebicyclo[7,2,0]undec-4-ene), found in various plants, is a natural bicyclic sesquiterpene with a low toxicity. Here, we show that a single intraperitoneal injection of β-caryophyllene (10 mg/kg) significantly reduced the cortical infarct volume by 67% when given immediately before middle cerebral artery occlusion (MCAO). Neurological deficits caused by MCAO were also significantly decreased by β-caryophyllene. β-Caryophyllene treatment of cortical cells exposed to oxygen-glucose deprivation revealed a significant protection in a dose-dependent manner. However, β-caryophyllene neither suppressed N-methyl-d-aspartate excitotoxicity in cultured cortical cells nor markedly decreased the oxidative stress measured in the cellular or acellular systems. By contrast, treatments with β-caryophyllene dose-dependently inhibited mRNA expression of inducible nitric oxide synthetase, interleukin (IL)-1β, IL-6, and cyclooxygenase 2 in C6 microglial cells, and decreased the level of nitric oxide and prostaglandin E2 at a 100 μM concentration. All of these findings suggest that β-caryophyllene has a potent neuroprotective activity, and its neuroprotection may be partly related to the modulation of inflammatory mediators.
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The present review discusses clinically important evidence for unstable remission/relapse patterns and for causes of premature mortality in patients with epilepsy. Although two of three patients become seizure-free with antiepileptic drugs (AEDs), as many as 20-40% of newly diagnosed patients with epilepsy become seizure-free without or with minimal AED treatment. Sadly, the proportion of those not becoming seizure-free despite treatment does not seem to have improved substantially over the past 50 years. There is emerging evidence that natural history of epilepsies is more complex than previously thought. Although two-thirds of patients follow stable patterns, that is, become seizure-free early and remain seizure-free (48%), or have refractory epilepsy all their life (19%), one in every three patients has an unstable course of epilepsy. Patients with an unstable course will either enter remission only after many years of having seizures (19%) or will relapse despite continued treatment (14%). Remote symptomatic epilepsy carries the highest risk of premature mortality. It remains unclear, however, why sudden unexplained death occurs more often in adults than in children and controversy exists whether entering remission prevents premature mortality. Studies of the natural history of newly diagnosed epilepsy show unexplained fluctuations of remission and relapse in as many as one in three patients and, sadly, no substantial improvement of epilepsy treatment in the past 50 years. The premature mortality of epilepsy is high and more work is needed to prevent it.
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The psychoactive cannabinoids from Cannabis sativa L. and the arachidonic acid-derived endocannabinoids are nonselective natural ligands for cannabinoid receptor type 1 (CB1) and CB2 receptors. Although the CB1 receptor is responsible for the psychomodulatory effects, activation of the CB2 receptor is a potential therapeutic strategy for the treatment of inflammation, pain, atherosclerosis, and osteoporosis. Here, we report that the widespread plant volatile (E)-β-caryophyllene [(E)-BCP] selectively binds to the CB2 receptor (K i = 155 ± 4 nM) and that it is a functional CB2 agonist. Intriguingly, (E)-BCP is a common constituent of the essential oils of numerous spice and food plants and a major component in Cannabis. Molecular docking simulations have identified a putative binding site of (E)-BCP in the CB2 receptor, showing ligand π–π stacking interactions with residues F117 and W258. Upon binding to the CB2 receptor, (E)-BCP inhibits adenylate cylcase, leads to intracellular calcium transients and weakly activates the mitogen-activated kinases Erk1/2 and p38 in primary human monocytes. (E)-BCP (500 nM) inhibits lipopolysaccharide (LPS)-induced proinflammatory cytokine expression in peripheral blood and attenuates LPS-stimulated Erk1/2 and JNK1/2 phosphorylation in monocytes. Furthermore, peroral (E)-BCP at 5 mg/kg strongly reduces the carrageenan-induced inflammatory response in wild-type mice but not in mice lacking CB2 receptors, providing evidence that this natural product exerts cannabimimetic effects in vivo. These results identify (E)-BCP as a functional nonpsychoactive CB2 receptor ligand in foodstuff and as a macrocyclic antiinflammatory cannabinoid in Cannabis. • Cannabis • CB2 cannabinoid receptor • foodstuff • inflammation • natural product
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CB 2 receptors Conditioned place preference LORR Two-bottle choice Several recent studies have suggested that brain CB 2 cannabinoid receptors play a major role in alcohol reward. In fact, the implication of cannabinoid neurotransmission in the reinforcing effects of ethanol (EtOH) is becoming increasingly evident. The CB 2 receptor agonist, β-caryophyllene (BCP) was used to investigate the role of the CB 2 receptors in mediating alcohol intake and ethanol-induced conditioned place preference (EtOH-CPP) and sensitivity in mice. The effect of BCP on alcohol intake was evaluated using the standard two-bottle choice drinking method. The mice were presented with increasing EtOH concentrations and its consumption was measured daily. Consumption of saccharin and quinine solutions was measured following the EtOH preference tests. Finally, the effect of BCP on alcohol reward and sensitivity was tested using an unbiased EtOH-CPP and loss of righting-reflex (LORR) procedures, respectively. BCP dose-dependently decreased alcohol consumption and preference. Additionally, BCP-injected mice did not show any difference from vehicle mice in total fluid intake in a 24-hour paradigm nor in their intake of graded concentrations of saccharin or quinine, suggesting that the CB 2 receptor activation did not alter taste function. More importantly, BCP inhibited EtOH-CPP acquisition and exacerbated LORR duration. Interestingly, these effects were abrogated when mice were pre-injected with a selective CB 2 receptor antagonist, AM630. Overall, the CB 2 receptor system appears to be involved in alcohol dependence and sensitivity and may represent a potential pharmacological target for the treatment of alcoholism.
Article
Trans-caryophyllene (TC), a component of essential oil found in many flowering plants, has shown its neuroprotective effects in various neurological disorders. However, the effects of TC on epilepsy haven't been reported before. In this study, we investigated the effect of TC on kainic acid-induced seizure activity caused by oxidative stress and pro-inflammation. We found that TC pretreatment significantly decreased seizure activity score compared to kainic acid treated group. Importantly, TC pretreatment leads to lowering the mortality in kainic acid treated mice. In addition, TC was found to significantly inhibit KA-induced generation of malondialdehyde. TC pretreatment also preserved the activity of GPx, SOD, and CAT. Notably, our data shows that an important property of TC is its capacity to exert cerebral anti-inflammatory effects by mitigating the expression of proinflammatory cytokines, such as TNF-α and IL-1β. These data suggest that TC has a potential protective effect on chemical induced seizure and brain damage.
Article
β-caryophyllene (BCP), a natural bicyclic sesquiterpene present in several essential oils, displays analgesic and anti-inflammatory properties in vitro and in vivo. Astrocytes are a major class of glial cells that regulate extracellular ion balance, repair and scarring processes in the central nervous system (CNS) following neuroinflammatory conditions and traumatic injuries. This study sought to determine the protective effect of BCP against glutamate-induced cytotoxicity in the C6 glioma cell line on neurochemical parameters as well as their biochemical mechanism. Glutamate increases intracellular ROS production and induces mitochondrial dysfunction as well as decreasing antioxidant defenses such as GSH and GPx activity. BCP prevented C6 cells from glutamate-induced cytotoxicity by modulating the cellular antioxidant response, mainly by inhibiting ROS production and reestablishing the mitochondrial membrane potential (Δψm). Moreover, BCP per se induced the nuclear translocation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) which was reflected by improvement in the cellular GSH antioxidant system. Taken together, our results suggest that cytoprotective effects of BCP were mediated by the amelioration of cellular antioxidant responses via Nrf-2 activation, which is, in part, dependent of CB2R activation. This functional nonpsychoactive CB2R ligand, could represent an important molecule for protection of glial cells against oxidative stress induced by glutamate.
Article
Background/aims: The activation of cannabinoid receptor 2 (CB2) has the beneficial effect of reducing neuroinflammatory response in the treatment of Alzheimer's disease (AD) and is suggested to trigger the peroxisome proliferator-activated receptor-γ (PPARγ) pathway; agonists of both receptors improve AD. Recently, the plant metabolite β-caryophyllene was shown to selectively bind to CB2 receptor and act as a full agonist. Methods: In this study, we examined the anti-inflammatory effect of β-caryophyllene in a transgenic APP/PS1 AD model and analyzed whether this effect was mediated by CB2 and PPARγ. Results: β-Caryophyllene, given orally, prevented cognitive impairment in APP/PS1 mice, and this positive cognitive effect was associated with reduced β-amyloid burden in both the hippocampus and the cerebral cortex. Moreover, β-caryophyllene reduced astrogliosis and microglial activation as well as the levels of COX-2 protein and the mRNA levels of the proinflammatory cytokines tumor necrosis factor-α and interleukin-1β in the cerebral cortex. The use of the CB2 antagonist AM630 or the PPARγ antagonist GW9662 significantly reversed the protective effects of β-caryophyllene on APP/PS1 mice. Conclusion: These results demonstrate that the anti-inflammatory effect of the sesquiterpene β-caryophyllene involves CB2 receptor activation and the PPARγ pathway and suggest β-caryophyllene as an attractive molecule for the development of new drugs with therapeutic potential for the treatment of AD.
Article
Several recent studies have suggested that brain CB2 cannabinoid receptors play a major role in alcohol reward. In fact, the implication of cannabinoid neurotransmission in the reinforcing effects of ethanol (EtOH) is becoming increasingly evident. The CB2 receptor agonist, β-Caryophyllene (BCP) was used to investigate the role of the CB2 receptors in mediating alcohol intake and ethanol-induced conditioned place preference (EtOH-CPP) and sensitivity in mice. The effect of BCP on alcohol intake was evaluated using the standard two-bottle choice drinking method. The mice were presented with increasing EtOH concentrations and its consumption was measured daily. Consumption of saccharin and quinine solutions was measured following the EtOH preference tests. Finally, the effect of BCP on alcohol reward and sensitivity was tested using an unbiased EtOH-CPP and loss of righting-reflex (LORR) procedures, respectively. BCP dose dependently decreased alcohol consumption and preference. Additionally, BCP-injected mice did not show any difference from vehicle mice in total fluid intake in a 24-hour paradigm nor in their intake of graded concentrations of saccharin or quinine, suggesting that the CB2 receptor activation did not alter taste function. More importantly, BCP inhibited EtOH-CPP acquisition and exacerbated LORR duration. Interestingly, these effects were abrogated when mice were pre-injected with a selective CB2 receptor antagonist, AM630. Overall, the CB2 receptor system appears to be involved in alcohol dependence and sensitivity and may represent a potential pharmacological target for the treatment of alcoholism.
Article
Epilepsy was defined conceptually in 2005 as a disorder of the brain characterized by an enduring predisposition to generate epileptic seizures. This definition is usually practically applied as having two unprovoked seizures >24 h apart. The International League Against Epilepsy (ILAE) accepted recommendations of a task force altering the practical definition for special circumstances that do not meet the two unprovoked seizures criteria. The task force proposed that epilepsy be considered to be a disease of the brain defined by any of the following conditions: (1) At least two unprovoked (or reflex) seizures occurring >24 h apart; (2) one unprovoked (or reflex) seizure and a probability of further seizures similar to the general recurrence risk (at least 60%) after two unprovoked seizures, occurring over the next 10 years; (3) diagnosis of an epilepsy syndrome. Epilepsy is considered to be resolved for individuals who either had an age-dependent epilepsy syndrome but are now past the applicable age or who have remained seizure-free for the last 10 years and off antiseizure medicines for at least the last 5 years. “Resolved” is not necessarily identical to the conventional view of “remission or “cure.” Different practical definitions may be formed and used for various specific purposes. This revised definition of epilepsy brings the term in concordance with common use.A PowerPoint slide summarizing this article is available for download in the Supporting Information section here.
Adverse effects of antiepileptic drugs (AEDs) are common, can have a considerable impact on quality of life and contribute to treatment failure in up to 40% of patients. The adverse effect profiles of AEDs differ greatly and are often a determining factor in drug selection because of the similar efficacy rates shown by most AEDs. The most common adverse effects are dose dependent and reversible. Cognitive impairment is of particular concern, especially for patients who work or study. Idiosyncratic effects, such as skin rashes, and chronic effects, such as weight gain, can lead to high rates of treatment discontinuation and complicate clinical management. Nearly all conventional AEDs increase the risk of congenital malformations when taken during pregnancy, with valproate posing a potentially greater risk, whereas the potential teratogenicity of new generation AEDs is largely unknown. Most conventional AEDs have a poor record when it comes to drug interactions, largely because of their tendency to interfere with hepatic drug metabolism. Some newer AEDs have no effect on hepatic drug metabolizing enzymes and are renally excreted, resulting in a lower potential for drug interactions. However, further research is needed to confirm the apparent improvement in tolerability offered by some of the newer AEDs.
Article
The type 2 cannabinoid receptor (CB2R) was recently shown to mediate neuroprotection in ischemic injury. However, the role of CB2Rs in the central nervous system, especially neuronal and glial CB2Rs in the cortex, remains unclear. We, therefore, investigated anti-ischemic mechanisms of cortical CB2R activation in various ischemic models. In rat cortical neurons/glia mixed cultures, a CB2R agonist, trans-caryophyllene (TC), decreased neuronal injury and mitochondrial depolarization caused by oxygen-glucose deprivation/re-oxygenation (OGD/R); these effects were reversed by the selective CB2R antagonist, AM630, but not by a type 1 cannabinoid receptor antagonist, AM251. Although it lacked free radical scavenging and antioxidant enzyme induction activities, TC reduced OGD/R-evoked mitochondrial dysfunction and intracellular oxidative stress. Western blot analysis demonstrated that TC enhanced phosphorylation of AMP-activated protein kinase (AMPK) and cAMP responsive element-binding protein (CREB), and increased expression of the CREB target gene product, brain-derived neurotrophic factor. However, TC failed to alter the activity of either Akt or extracellular signal-regulated kinase, two major CB2R signaling pathways. Selective AMPK and CREB inhibitors abolished the neuroprotective effects of TC. In rats, post-ischemic treatment with TC decreased cerebral infarct size and edema, and increased phosphorylated CREB and brain-derived neurotrophic factor expression in neurons. All protective effects of TC were reversed by co-administration with AM630. Collectively, these data demonstrate that cortical CB2R activation by TC ameliorates ischemic injury, potentially through modulation of AMPK/CREB signaling, and suggest that cortical CB2Rs might serve as a putative therapeutic target for cerebral ischemia.
Article
Background: β-caryophyllene (BCP) is a common constitute of the essential oils of numerous spice, food plants and major component in Cannabis. The present study investigated the contribution of peripheral cannabinoid (CB) and opioid systems in the antinociception produced by intraplantar (i.pl.) injection of BCP. The interaction between peripheral BCP and morphine was also examined. Methods: The antinociceptive effect of i.pl. BCP was assayed by the capsaicin tests in mice. Antagonists for CB and opioid receptors, and antisera against β-endorphin were injected peripherally prior to i.pl. injection of BCP. Morphine in combination with BCP was injected subcutaneously or intrathecally. Results: The i.pl. injection of BCP dose-dependently attenuated capsaicin-induced nociceptive response. The antinociceptive effect produced by BCP was prevented by pretreatment with AM630, a selective CB2 receptor antagonist, but not by AM251, a selective CB1 receptor antagonist. Pretreatment with naloxone, an opioid receptor antagonist, and β-funaltrexamine, a selective μ-opioid receptor antagonist, reversed the antinociceptive effect of BCP. Pretreatment with naloxone methiodide, a peripherally acting antagonist for opioid receptors and antisera against β-endorphin, resulted in a significant antagonizing effect on BCP-induced antinociception. Morphine-induced antinociception was increased by a low dose of BCP. The increased effect of morphine in combination with BCP was antagonized significantly by pretreatment with naloxone. Conclusions: The present results demonstrate that antinociception produced by i.pl. BCP is mediated by activation of CB2 receptors, which stimulates the local release from keratinocytes of the endogenous opioid β-endorphin. The combined injection of morphine and BCP may be an alternative in treating chemogenic pain.
Article
Zearalenone (ZEA) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium, commonly found in the soil in temperate and warm countries and is a frequent contaminant of cereal crops worldwide. Accordingly, it has been implicated in several mycotoxicosis in farm animals and in humans, but the underlying mechanisms remain largely unknown. Therefore, the current study was aimed to investigate the effect of an acute dose of ZEA (40 mg/kg, p.o.) on reproductive and hematological parameters, as well as on markers of oxidative stress in liver, kidney and testes in mice. Adult Swiss albino male mice were exposed to a single oral administration of ZEA, and 48 h thereafter behavioral and biochemical tests were performed. No differences in locomotor or exploratory activity were observed in the open-field test. On the other hand, ZEA increased the number of leukocytes, segmented neutrophils, sticks, eosinophils, monocytes and decreased platelets and lymphocytes number. Moreover, ZEA drastically reduced the number and motility of live spermatozoa. Additionally, while levels of thiobarbituric acid reactive substances (TBARS), non-protein thiols (NPSH) and ascorbic acid in liver, kidney and testes were not altered by ZEA administration, superoxide dismutase activity increased in all tissues evaluated, catalase activity increased in the kidney, and glutathione-S-transferase activity decreased in kidney and testes. In summary, we showed that ZEA have acute toxic effects mainly in reproductive system of adult male Swiss albino mice and its effect probably is related to a reduced activity of GST and increased in SOD activity in testes.
Article
Animal models of memory have been considered as the subject of many scientific publications at least since the beginning of the twentieth century. In humans, memory is often accessed through spoken or written language, while in animals, cognitive functions must be accessed through different kind of behaviors in many specific, experimental models of memory and learning. Among them, the novel object recognition test can be evaluated by the differences in the exploration time of novel and familiar objects. Its application is not limited to a field of research and enables that various issues can be studied, such as the memory and learning, the preference for novelty, the influence of different brain regions in the process of recognition, and even the study of different drugs and their effects. This paper describes the novel object recognition paradigms in animals, as a valuable measure of cognition. The purpose of this work was to review the neurobiology and methodological modifications of the test commonly used in behavioral pharmacology.
Article
Animal models for seizures and epilepsy have played a fundamental role in advancing our understanding of basic mechanisms underlying ictogenesis and epileptogenesis and have been instrumental in the discovery and preclinical development of novel antiepileptic drugs (AEDs). However, there is growing concern that the efficacy of drug treatment of epilepsy has not substantially improved with the introduction of new AEDs, which, at least in part, may be due to the fact that the same simple screening models, i.e., the maximal electroshock seizure (MES) and s.c. pentylenetetrazole (PTZ) seizure tests, have been used as gatekeepers in AED discovery for >6 decades. It has been argued that these old models may identify only drugs that share characteristics with existing drugs, and are unlikely to have an effect on refractory epilepsies. Indeed, accumulating evidence with several novel AEDs, including levetiracetan, has shown that the MES and PTZ models do not identify all potential AEDs but instead may fail to discover compounds that have great potential efficacy but work through mechanisms not tested by these models. Awareness of the limitations of acute seizure models comes at a critical crossroad. Clearly, preclinical strategies of AED discovery and development need a conceptual shift that is moving away from using models that identify therapies for the symptomatic treatment of epilepsy to those that may be useful for identifying therapies that are more effective in the refractory population and that may ultimately lead to an effective cure in susceptible individuals by interfering with the processes underlying epilepsy. To realize this goal, the molecular mechanisms of the next generation of therapies must necessarily evolve to include targets that contribute to epileptogenesis and pharmacoresistance in relevant epilepsy models.
Article
Beta-caryophyllene (BC) is a chemical found in the essential oil of numerous plants which have shown various biomedical properties. In the present investigation we examined for the first time its genotoxic capacity in vivo by evaluating its potential to induce micronuclei in mouse. In a first assay we administered three doses of the compound once, and in a second assay, we administered three doses of BC in three consecutive days. In each assay, the results were compared with those obtained in a group of mice administered corn oil and in another treated with adryamicin. Our results revealed no genotoxic effect by BC in the two assays. Moreover, no bone marrow cyotoxicity was observed when the proportion of polychromatic erythrocytes was determined with respect to the number of normochromatic erythrocytes. Therefore, our study showed the relevance of extending research on BC regarding its beneficial properties.
Article
The International League Against Epilepsy (ILAE) and the International Bureau for Epilepsy (IBE) have come to consensus definitions for the terms epileptic seizure and epilepsy. An epileptic seizure is a transient occurrence of signs and/or symptoms due to abnormal excessive or synchronous neuronal activity in the brain. Epilepsy is a disorder of the brain characterized by an enduring predisposition to generate epileptic seizures and by the neurobiologic, cognitive, psychological, and social consequences of this condition. The definition of epilepsy requires the occurrence of at least one epileptic seizure.
Article
Synaptic activity and ischemia/injury promote lipid messenger formation through phospholipase-mediated cleavage of specific phospholipids from membrane reservoirs. Lipid messengers modulate signaling cascades, contributing to development, differentiation, function (e.g., memory), protection, regeneration, and repair of neurons and overall regulation of neuronal, glial, and endothelial cell functional integrity. Oxidative stress disrupts lipid signaling and promotes lipid peroxidation and neurodegeneration. Lipid signaling at the neurovascular unit (neurons, astrocytes, oligodendrocytes, microglia, and cells of the microvasculature) is altered in early cerebrovascular and neurodegenerative disease. We discuss how lipid signaling regulates critical events in neuronal survival. Aberrant synaptic plasticity (e.g., epileptogenesis) is highlighted to show how gene expression may drive synaptic circuitry formation in the "wrong" direction. Docosahexaenoic acid has been implicated in memory, photoreceptor cell biogenesis and function, and neuroprotection. Free docosahexaenoic acid released in the brain during experimental stroke leads to the synthesis of stereospecific messengers through oxygenation pathways. One messenger, 10,17S-docosatriene (neuroprotectin D1; NPD1), counteracts leukocyte infiltration and proinflammatory gene expression in brain ischemia-reperfusion. In retina, photoreceptor survival depends on retinal pigment epithelial (RPE) cell integrity. NPD1 is synthesized in RPE cells undergoing oxidative stress, potently counteracts oxidative stress-triggered apoptotic DNA damage in RPE, upregulates antiapoptotic proteins Bcl-2 and Bcl-x(L), and decreases proapoptotic Bax and Bad expression. These findings expand our understanding of how the nervous system counteracts redox disturbances, mitochondrial dysfunction, and proinflammatory conditions. The specificity and potency of NPD1 indicate a potential target for therapeutic intervention for stroke, age-related macular degeneration, spinal cord injury, and other neuroinflammatory or neurodegenerative diseases.
Article
Oxidative stress may contribute to epileptogenicity in genetic models of epilepsy. To address this, we examined the enzymatic activity of cytosolic Cu/Zn superoxide dismutase (SOD-1), mitochondrial Mn superoxide dismutase (SOD-2), and glutathione peroxidase (GPx) in the developing hippocampus of genetically epilepsy-prone rats (GEPR-9s). We also measured changes in the GSH/GSSG ratio, lipid peroxidation, and protein oxidation at post-natal days (PD) 7, 30, and 90, respectively. Compared with control Sprague-Dawley (SD) rats, GEPR-9s showed similar SOD-1 and SOD-2 activity but lower GPx activity. Epilepsy-prone rats also showed lower GSH/GSSG ratios than controls, and more lipid peroxidation (as measured by malondialdehyde levels) and protein oxidation (as measured by carbonyl levels). Treatment with kainic acid (KA) resulted in more pronounced seizures, less GPx activity, and lower GSH/GSSG ratios in GEPR-9s than in controls, but KA did not significantly affect SOD-1 or SOD-2 activity, suggesting that GEPR-9s do not compensate for reduced GPx activity by increasing SOD. Moreover, KA treatment resulted in significantly a lower GSH/GSSG ratio and GPx-like immunoreactivity and higher malondialdehyde and carbonyl levels in GEPR-9s than in controls. These findings were more evident in GEPR-9s at PD 90 than at PD 30, indicating that oxidative stress is age-dependent. Double-labeling immunocytochemical analysis demonstrated co-localization of GPx-immunoreactive glia-like cells and reactive astrocytes, as labeled by glial fibrillary acidic protein (GFAP). This suggests that mobilization of astroglial cells for synthesis of GPx protein is a response to KA insult, intended to decrease the neurotoxicity induced by peroxides. These responses were more pronounced in control SD rats than in GEPR-9s. Our results suggest that impairment of the GPx (including glutathione)-mediated antioxidant system contributed to epileptogenesis in GEPR-9s.