Caryophyllene Research Articles

Beta-Caryophyllene Exhibits Anti-Proliferative Effects through Apoptosis Induction and Cell Cycle Modulation in Multiple Myeloma Cells

Simple SummaryMultiple myeloma (MM) is a malignant B-cell neoplasm characterized by the uncontrolled proliferation of plasma cells. MM cells highly express cannabinoid type 2 receptors (CB2Rs), and previous studies have already demonstrated that the Cannabis plant and its derivatives may have anti-emetic as well as anti-neoplastic effects. In the present study, β-caryophyllene (BCP), a natural CB2R agonist, was evaluated for its anti-proliferative and anti-cancer effects. BCP was able to induce the apoptotic mechanism by activating the molecules involved in triggering apoptosis, such as Bax and caspase 3, and it reduced the anti-apoptotic protein Bcl-2; BCP also regulated cell proliferation through sophisticated crosstalk between Akt, β-catenin, and cyclin D1/CDK 4-6 in a concentration-dependent manner. These effects were counteracted by AM630, a CB2R antagonist, thus showing that BCP acts through CB2R. The data obtained so far demonstrate that BCP, thanks to its anti-proliferative effects, might represent an interesting additional therapeutic approach to improve anti-myeloma therapy. Cannabinoid receptors, which are widely distributed in the body, have been considered as possible pharmacological targets for the management of several tumors. Cannabinoid type 2 receptors (CB2Rs) belong to the G protein-coupled receptor family and are mainly expressed in hematopoietic and immune cells, such as B-cells, T-cells, and macrophages; thus, CB2R activation might be useful for treating cancers affecting plasma cells, such as multiple myeloma (MM). Previous studies have shown that CB2R stimulation may have anti-proliferative effects; therefore, the purpose of the present study was to explore the antitumor effect of beta-caryophyllene (BCP), a CB2R agonist, in an in vitro model of MM. Dexamethasone-resistant (MM.1R) and sensitive (MM.1S) human multiple myeloma cell lines were used in this study. Cells were treated with different concentrations of BCP for 24 h, and a group of cells was pre-incubated with AM630, a specific CB2R antagonist. BCP treatment reduced cell proliferation through CB2R stimulation; notably, BCP considerably increased the pro-apoptotic protein Bax and decreased the anti-apoptotic molecule Bcl-2. Furthermore, an increase in caspase 3 protein levels was detected following BCP incubation, thus demonstrating its anti-proliferative effect through apoptosis activation. In addition, BCP regulated AKT, Wnt1, and beta-catenin expression, showing that CB2R stimulation may decrease cancer cell proliferation by modulating Wnt/β-catenin signaling. These effects were counteracted by AM630 co-incubation, thus confirming that BCP’s mechanism of action is mainly related to CB2R modulation. A decrease in β-catenin regulated the impaired cell cycle and especially promoted cyclin D1 and CDK 4/6 reduction. Taken together, these data revealed that BCP might have significant and effective anti-cancer and anti-proliferative effects in MM cells by activating apoptosis, modulating different molecular pathways, and downregulating the cell cycle.

ANTIOXIDANT ACTIVITY, SKIN IRRITAION POTENTIAL AND CHEMICAL COMPOSITION OF CLOVE LEAF OIL FROM WEST JAVA INDONESIA

ANTIOXIDANT ACTIVITY, SKIN IRRITATON POTENTIAL, AND CHEMICAL COMPOSITION OF CLOVE LEAF OIL FROM WEST JAVA INDONESIA. Essential oils with specific chemical compositions have the potential as a source of antioxidants in cosmetics, however, in general, essential oils are not safe because of the potential for skin irritation. This study aims is to perform fractionation to obtain clove oil with a chemical composition that is safe on the skin and does not reduce its antioxidant activity. Fractionation was performed at a temperature of 100 – 285 oC. Chemical composition was determined by Mass Chromatography Spectrometry (GC-MS), antioxidant activity by DPPH method, and irritation potential by in-vivo patch test. The results showed chemical components of clove oil were eugenol, copaene, beta caryophyllene, iso-eugenol, alpha caryophyllene, cadinene, caryophyllene oxide, caryophylla and beta caryophylla. Fractions with chemical compositions of eugenol and beta caryophyllene as the main components and copaene, iso-eugenol, alpha caryophyllene, cadinene as secondary components have very light erythema and no potential irritation to the skin. The fraction produced from the fractionation of crude clove oil at a temperature of 235 – 260 oC has the highest antioxidant activity of 10.17 mg/L.

GC-MS/FID analysis, antibacterial activity and modes of action of essential oils from three Aframomum species found in Cameroon against foodborne pathogenic bacteria

Background: The current study examined the chemical profile and in vitro antibacterial activity of essential oils (EOs) extracted from Aframomum danielli (leaves and seeds), Aframomum chlamydentum (leaves), and Aframomum melegueta (leaves) against foodborne pathogenic bacteria.
 Methods: The hydro-distillation technique using a Clevenger-type apparatus was used to extract EOs, whereas the Gas Chromatography-Mass Spectrometry (GC-MS) and GC coupled to Flame Ionization Detector (GC-FID) allowed the chemical characterization of oil constituents. The broth micro-dilution method was applied for the determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Besides, some modes of action were studied on the cell membrane integrity and biofilm formation of Salmonella typhi.
 Results: The major compounds identified from EOs of A. danielli seeds were eucalyptol (48.707%), limonene (11.368%), beta pinene (10.342-10.335%), and alpha terpineol (8.785-9.049%), whereas EOs from A. danielli leaves were dominated by sabinene (42.87%), beta pinene, (11.22%), caryophyllene (7.84%), terpinen-4-ol (5.68%), linalool (3.48%) and gamma terpinene (2.02%). Major volatile markers from EOs of A. chlamydentum leaves comprised beta pinene (49.72%), caryophyllene (10.62%), alpha pinene (6.21%) and linalool (2.96%), while those of EOs from A. melegueta included beta pinene (37.15%), caryophyllene (17.64%), caryophyllene oxide (8.72%) and alpha pinene (8.26%). This study is the first to report on the chemical constituents of EOs from A. chlamydentum. Test oils displayed significant antibacterial activity with the MIC ranging from 0.0625 to 0.5% (v/v). EOs from A. melegueta (leaves) appeared to be the most active, acting against all tested bacteria. All EOs identified displayed bactericidal effects against Citrobacter freundii, a bacterium known to cause a broad range of infections associated with a higher rate of in-hospital mortality. The EOs from A. melegueta may act through perturbation of cell membrane integrity and permeability as well as the inhibition of bacterial biofilm formation.
 Conclusion: Our findings suggest the possible application of essential oils in agricultural food products for the control of bacterial diseases.

β-Caryophyllene Induces Apoptosis and Inhibits Angiogenesis in Colorectal Cancer Models.

Beta-Caryophyllene (BCP), a naturally occurring sesquiterpene abundantly found in cloves, hops, and cannabis, is the active candidate of a relatively new group of vascular-inhibiting compounds that aim to block existing tumor blood vessels. Previously, we have reported the anti-cancer properties of BCP by utilizing a series of in-vitro anti-tumor-related assays using human colorectal carcinoma cells. The present study aimed to investigate the effects of BCP on in-vitro, ex-vivo, and in-vivo models of anti-angiogenic assays and evaluate its anti-cancer activity in xenograft tumor (both ectopic and orthotopic) mice models of human colorectal cancer. Computational structural analysis and an apoptosis antibody array were also performed to understand the molecular players underlying this effect. BCP exhibited strong anti-angiogenic activity by blocking the migration of endothelial cells, tube-like network formation, suppression of vascular endothelial growth factor (VEGF) secretion from human umbilical vein endothelial cells and sprouting of rat aorta microvessels. BCP has a probable binding at Site#0 on the surface of VEGFR2. Moreover, BCP significantly deformed the vascularization architecture compared to the negative control in a chick embryo chorioallantoic membrane assay. BCP showed a remarkable reduction in tumor size and fluorescence molecular tomography signal intensity in all the mice treated with BCP, in a dose-dependent relationship, in ectopic and orthotopic tumor xenograft models, respectively. The histological analysis of the tumor from BCP-treated mice revealed a clear reduction of the density of vascularization. In addition, BCP induced apoptosis through downregulation of HSP60, HTRA, survivin, and XIAP, along with the upregulation of p21 expressions. These results suggest that BCP acts at multiple stages of angiogenesis and could be used as a promising therapeutic candidate to halt the growth of colorectal tumor cells.

Therapeutic potential of beta-caryophyllene against aflatoxin B1-Induced liver toxicity: biochemical and molecular insights in rats

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.

β-Caryophyllene, A Natural Dietary CB2 Receptor Selective Cannabinoid can be a Candidate to Target the Trinity of Infection, Immunity, and Inflammation in COVID-19.

Coronavirus disease (COVID-19), caused by novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is an ongoing pandemic and presents a public health emergency. It has affected millions of people and continues to affect more, despite tremendous social preventive measures. Identifying candidate drugs for the prevention and treatment of COVID-19 is crucial. The pathogenesis and the complications with advanced infection mainly involve an immune-inflammatory cascade. Therefore, therapeutic strategy relies on suppressing infectivity and inflammation, along with immune modulation. One of the most promising therapeutic targets for the modulation of immune-inflammatory responses is the endocannabinoid system, particularly the activation of cannabinoid type 2 receptors (CB2R), a G-protein coupled receptor which mediates the anti-inflammatory properties by modulating numerous signaling pathways. To pharmacologically activate the CB2 receptors, a naturally occurring cannabinoid ligand, beta-caryophyllene (BCP), received attention due to its potent anti-inflammatory, antiviral, and immunomodulatory properties. BCP is recognized as a full selective functional agonist on CB2 receptors and produces therapeutic effects by activating CB2 and the nuclear receptors, peroxisome proliferator-activated receptors (PPARs). BCP is regarded as the first dietary cannabinoid with abundant presence across cannabis and non-cannabis plants, including spices and other edible plants. BCP showed tissue protective properties and favorably modulates numerous signaling pathways and inhibits inflammatory mediators, including cytokines, chemokines, adhesion molecules, prostanoids, and eicosanoids. Based on its pharmacological properties, molecular mechanisms, and the therapeutic potential of BCP as an immunomodulator, anti-inflammatory, organ-protective, and antiviral, we hypothesize that BCP could be a promising therapeutic and/or preventive candidate to target the triad of infection, immunity, and inflammation in COVID-19. In line with numerous studies that proposed the potential of cannabinoids in COVID-19, BCP may be a novel candidate compound for pharmaceutical and nutraceutical development due to its unique functional receptor selectivity, wide availability and accessibility, dietary bioavailability, nonpsychoactivity, and negligible toxicity along with druggable properties, including favorable pharmacokinetic and physicochemical properties. Based on reasonable pharmacological mechanisms and therapeutic properties, we speculate that BCP has potential to be investigated against COVID-19 and will inspire further preclinical and clinical studies.

In Vitro Evaluation of the Activity of Terpenes and Cannabidiol against Human Coronavirus E229.

The activity of a new, terpene-based formulation, code-named NT-VRL-1, against Human Coronavirus (HCoV) strain 229E was evaluated in human lung fibroblasts (MRC-5 cells), with and without the addition of cannabidiol (CBD). The main constituents in the terpene formulation used for the experiment were beta caryophyllene, eucalyptol, and citral. The tested formulation exhibited an antiviral effect when it was pre-incubated with the host cells prior to virus infection. The combination of NT-VRL-1 with CBD potentiated the antiviral effect better than the positive controls pyrazofurin and glycyrrhizin. There was a strong correlation between the quantitative results from a cell-viability assay and the cytopathic effect seen under the microscope after 72 h. To the best of our knowledge, this is the first report of activity of a combination of terpenes and CBD against a coronavirus.

Chemical Characterization of Clove (Syzygium aromaticum) Bud Essential Oil and Microbicidal Activity against Isolated Clinical Strains

Clove (Syzygium aromaticum) is one of the most valuable spices that had been used for centuries as food preservative and for many medicinal purposes. So far, the studies on utilization of clove oil as an essential oil are still limited. In this study Syzygium aromaticum (clove) essential oil was isolated from buds by hydro-distillation process and characterized by GC-MS. Micorbiocidal and antioxidant potential was evaluated by agar well diffusion assay, DPPH assay and ferrous chelation potential assay. Major components identified were Eugenol (82.414%), Eugenyl acetate (15.196%), Beta caryophyllene (1.919%) and Alpha-caryophyllene (0.472%). Antimicrobial activity was evaluated against reference strains and clove oil exhibited potential antifungal activity against Bipolaris specifera, Curvularia lunata and Alternaria alternata with IC50 value 0.014 mg/ml, 0.015 mg/ml and 0.014 mg/ml (w/v), respectively and antibacterial activity against Bacillus cereus (24mm), Bacillus subtilis (6mm), Micrococcus luteus (15mm), Alcalygens denitrificans (13mm), Pseudomonas alcaligens (9mm) and Enterococcus fecalis (12mm) but no activity was observed against Camphylobacter coli and Klebsiella pneumonia. Further, microbiocidal properties against clinically isolated bacterial strains were also analyzed and significant activity with MIC value of 0.125 to 0.625 mg/ml for Escherichia coli and 0.5±0.23 mg/ml (w/v) for Staphylococcus aureus was observed. Simultaneously, potential free radical scavenging 1.590±0.07 mg/ml (w/v) and ferrous chelation potential 11±1.2% was also noted. In conclusion, clove essential oil can be considered as a potential antimicrobial agent for food and pharmaceutical industries.

Mitochondrial Metabolism, Dysfunctions in Senescence Cell and the Possible Interventions through Herbal Medicines

The mitochondria are the cell`s powerhouse. They are considered ubiquitous organelles of all eukaryotic cells, being responsible for the cell’s life and death cycle. Through stimuli in the environment in which they live, mitochondria can modulate their own biogenesis as well as signal retrograde to the nucleus to modify the structure of their proteins. Since the mitochondrial genome contains only 37 genes, much of the encoding of its proteins depends on the nuclear genome. Thus, the communication between mitochondria and the nucleus seems to be a target of science in understanding the pathologies associated with this organelle. Some medicinal herbs have been shown to influence mitochondrial biogenesis, such as Gynostemma pentaphyllun (GP) and berberine, which increase the phosphorylation of proteins AMPactivated protein kinase (AMPK). Just as GP and berberine phosphorylate AMPK in signaling for mitochondrial biogenesis, the sesquiterpene beta-caryophyllene (BCP) demonstrated positive results in reorganizing mitochondrial transcription factors, being an agonist of the peroxisome proliferatoractivated alpha receptor (PPAR-α). Another plant derivative, the non-psychoactive cannabinoid known as cannabidiol (CBD), has been showing control in the metabolism of calcium in the mitochondrial matrix. In this review, we seek to get a closer look at the biochemical mechanisms of action of some of these plants, as well as their synergies in the results of different treatments. In the view of oriental medicines, the use of associated medicinal herbs has always been part of their treatment protocols. However, the effectiveness of these treatments in relation to plant synergy can be observed in future clinical trials for better understanding.

Composición de aceites esenciales de diferentes especies de “pimienta” de los géneros Piper, Pimenta, Lindera, Ruta, Schinus y Zanthoxylum

Se ha extraído mediante hidrodestilación el aceite esencial de diez especies usadas como pimienta: Piper borbonense, P. capense, P. retrofractum, P. nigrum, Zanthoxylum bungeanum y Z. armatum, Lindera neesiana, Ruta chalepensis, Schinus terebenthifolia, Pimenta dioica. Los análisis realizados mediante cromatografía de gases acoplada a espectrometría de masas encontraron que todas presentan β-felandreno y derivados de cariofileno y felandreno, siendo estos compuestos de propiedades pungentes los característicos de la especia pimienta. El rendimiento de esencia varía desde 0,43% para R. chalepensis hasta 7,61% para P. borbonense. Los compuestos mayoritarios fueron: P. borbonense (α-felandreno, 12,43%), P. capense (δ-cadineno, 25,59%,), P. retrofractum (γ-cadineno, 31,63%), P. nigrum ((E)-β-cariofileno, 22,88%), P. dioica (eugenol, 48,93%), L. neesiana (miristicina, 14,13%), R. chalepensis (2-undecanona, 64,93%), S. therebenthifolia (δ-3-careno, 29,21%), Z. armatum (linalool, 53,30%); Z. bungeanum (linalool, 64,09%). Todo esto muestra las diferencias en el metabolismo secundario de las pimientas y por tanto sus posibles aplicaciones en diferentes industrias.

Beta-caryophyllene inhibits cocaine addiction-related behavior by activation of PPARα and PPARγ: repurposing a FDA-approved food additive for cocaine use disorder.

Cocaine abuse continues to be a serious health problem worldwide. Despite intense research, there is still no FDA-approved medication to treat cocaine use disorder (CUD). In this report, we explored the potential utility of beta-caryophyllene (BCP), an FDA-approved food additive for the treatment of CUD. We found that BCP, when administered intraperitoneally or intragastrically, dose-dependently attenuated cocaine self-administration, cocaine-conditioned place preference, and cocaine-primed reinstatement of drug seeking in rats. In contrast, BCP failed to alter food self-administration or cocaine-induced hyperactivity. It also failed to maintain self-administration in a drug substitution test, suggesting that BCP has no abuse potential. BCP was previously reported to be a selective CB2 receptor agonist. Unexpectedly, pharmacological blockade or genetic deletion of CB1, CB2, or GPR55 receptors in gene-knockout mice failed to alter BCP's action against cocaine self-administration, suggesting the involvement of non-CB1, non-CB2, and non-GPR55 receptor mechanisms. Furthermore, pharmacological blockade of μ opioid receptor or Toll-like receptors complex failed to alter, while blockade of peroxisome proliferator-activated receptors (PPARα, PPARγ) reversed BCP-induced reduction in cocaine self-administration, suggesting the involvement of PPARα and PPARγ in BCP's action. Finally, we used electrical and optogenetic intracranial self-stimulation (eICSS, oICSS) paradigms to study the underlying neural substrate mechanisms. We found that BCP is more effective in attenuation of cocaine-enhanced oICSS than eICSS, the former driven by optical activation of midbrain dopamine neurons in DAT-cre mice. These findings indicate that BCP may be useful for the treatment of CUD, likely by stimulation of PPARα and PPARγ in the mesolimbic system.

Polygonum minus essential oil modulates cisplatin-induced hepatotoxicity through inflammatory and apoptotic pathways.

Oxidative stress, inflammation and apoptosis are thought as primary mediators of cisplatin-induced hepatotoxicity. The objective of this study was to determine the protective effect of Polygonum minus essential oil in cisplatin-induced hepatotoxicity. A total of forty-two male rats were randomly divided into seven groups: control, cisplatin, β-caryophyllene 150 mg/kg (BCP), PmEO 100 mg/kg + cisplatin (PmEO100CP), PmEO 200 mg/kg + cisplatin (PmEO200CP), PmEO 400 mg/kg + cisplatin (PmEO400CP) and PmEO 400 mg/kg (PmEO400). Rats in the BCP, PmEO100CP, PmEO200CP, PmEO400CP and PmEO400 group received respective treatment orally for 14 consecutive days prior to cisplatin injection. All animals except for those in the control group and PmEO400 were administered with a single dose of cisplatin (10 mg/kg) intraperitoneally on day 15 and all animals were sacrificed on day 18. PmEO100CP pretreatment protected against cisplatin-induced hepatotoxicity by decreasing CYP2E1 and indicators of oxidative stress including malondialdehyde, 8-OHdG and protein carbonyl which was accompanied by increased antioxidant status (glutathione, glutathione peroxidase, superoxide dismutase and catalase) as compared to cisplatin group. PmEO100CP pretreatment also modulated changes in liver inflammatory markers (TNF-α, IL-1α, IL-1β, IL-6 and IL-10). PmEO100CP administration also notably reduced cisplatin-induced apoptosis significantly as compared to cisplatin group. In conclusion, our results suggested that P. minus essential oil at a dose of 100 mg/kg may protect against cisplatin-induced hepatotoxicity possibly via inhibition of oxidative stress, inflammation and apoptosis.

In vitro and in vivo experimental trials to assess the modulatory influence of β-caryophyllene on NDV replication and immunopathogenesis

A trial was conducted to evaluate the antiviral activity and immunomodulatory effect of B-Caryophyllene (BCP) using NDV as a viral model. First, an in ovo experiment was conducted to estimate the antiviral mechanism of BCP. Next, an in vivo experiment was designed to confirm its antiviral efficacy as well as its immunomodulatory and growth promoting ability. According to the in ovo experiment, BCP possesses antiviral influence up to 61.7% when treated before or during NDV infection. Oral supplementation of chickens with two doses of BCP (200 and 400 μg/bird) resulted in a significant increase in the NDV HI-Ab responses and a significant increase in interferon-α signaling cytokines. These obvious immunomodulatory effects improved the bird clinical protection against virulent NDV challenge. To conclude, we introduced a new compound for the poultry industry sector that has antiviral and immunostimulant properties when supplemented orally before or during NDV infection.

Compositional Analysis of Volatile Compounds of Paspangiri - A Mixture of Leaves of Five Rutaceae Plants Used as an Effective Remedy for Respiratory System Microbial Infections in Traditional Systems of Medicine

Steaming with aromatic poly herbal formulations has been practiced as an effective treatment for curing or controlling infectious diseases, deterring of pest species, destroying of unwanted microbial contaminations by several cultures. Steaming of Pasppangiri, a mixture of leaves of five species (Citrus aurantium Linn, Citrus aurantifolia (Christm. & Panzer) Swingle, Citrus sinensis Linn, Atlantia ceylanica (Am.) Oliver, Citrus reticulata Blanco) has been used for an effective treatment for curing of microbial infected, respiratory system related diseases. However, information on volatile compounds, compositional analysis of Paspangiri is lacking or scattered. Leaves of five citrus species were collected from previously authenticated plants grown in same soil and climatic conditions. Essential oil was separated by hydro distillation method. The chemical composition of the essential oil of Paspangiri was analysed using gas chromatography-mass spectrometry (GC-MS). Results revealed that the number of compounds observed in each species were varied as Citrus sinensis (27)>aurantium (25) >Citrus reticulata (20)>Citrus aurantifolia (19)>Atlantia ceylanica (5) respectively. The major bioactive molecules identified in essential oils of Paspangiri such as linalool, limonene, caryophyllene, eugenol, z-citral, ∞- pinene, sabinene, and myrcene, either singularly or as a mixture have been exhibited antimicrobial properties. Therefore, the present study, partially validate the traditional claims of inhaling of Paspangiri steam for prevention or curing of infectious disease.

β-Caryophyllene Inhibits Cell Proliferation through a Direct Modulation of CB2 Receptors in Glioblastoma Cells.

Glioblastomas are aggressive cancers characterized by uncontrolled proliferation and inflammation. b-caryophyllene (BCP) is a cannabinoid receptor 2 (CB2) agonist that showed an important anti-inflammatory effect through the interaction of CB2 and peroxisome proliferator-activated receptor gamma (PPARg) receptors. BCP effects were investigated in an in vitro model of glioblastoma. U-373 and U87, derived from a human glioblastoma, and human glioma stem-like cells (GSCs) were treated with BCP at different doses and time-points. AM360, a specific CB2 antagonist, was added 2 h before BCP treatment. BCP showed a significant anti-proliferative effect, reducing cell viability, inhibiting cell cycle, and increasing apoptosis, as demonstrated by Tunel assay, caspase-3 and caspase -9 activation. In addition, the pro-apoptotic BAX expression was increased, whereas the anti-apoptotic Bcl-2 expression was reduced. Treatment with BCP decreased Beclin-1, LC3 and p62/SQSTM1 expression, indicating a possible switch of autophagy to apoptosis. BCP’s anti-inflammatory effect was demonstrated by NF-κB reduction, PPARg activation and TNF-a decrease; BCP significantly reduced Jun N-Terminal Kinase (JNK) expression as a consequence of TNF-α inhibition. AM360 abrogated BCP effects, thus demonstrating the BCP mechanism of action through the CB2 receptor. These findings let us hypothesize that BCP may act as a tumor suppressor in glioblastoma, acting on CB2 receptor and modulating JNK.

Beta‐caryophyllene, a Volatile Phytocannabinoid, Attenuates Cocaine Self‐administration and Relapse in Rats

Cocaine abuse continues to be a serious health problem worldwide. Despite intense research, there is still no FDA‐approved medication to treat cocaine use disorder. The recent search for effective therapeutics has been focused on the endocannabinoid system because of its critical role in the neurobiology of drug addiction. In the last decade, there has been a growing interest in beta‐caryophyllene (BCP), a volatile phytocannabinoid present in high proportions in cannabis and large number of spice and food plants including black pepper, thyme and cloves. This dietary additive that has a CB2 receptor (CB2R) agonist profile, has shown to produce promising therapeutic effects for multiple neuropsychiatric disorders. Surprisingly, the therapeutic potential of BCP in the treatment of drug abuse and addiction has not been explored. Here, using gold standard animal models of drug abuse, we systematically evaluated the potential utility of BCP against cocaine‐related behaviors in rodents and its mechanisms of action. In a series of experiments we found that BCP attenuated cocaine‐enhanced electrical brain‐stimulation reward and optogenetic intracranial self‐stimulation driven by activation of dopamine (DA) neurons in DAT‐cre mice, both outcomes indicating reduced reward. Intriguingly, when administered systemically or orally, BCP attenuated cocaine self‐administration in rats, suggesting an ability to reduce drug abuse. BCP also reduced drug‐primed reinstatement of cocaine seeking and cocaine conditioned place preference, indicative of its preventative effects against relapse. When BCP was substituted for cocaine, rats ceased responding, suggesting BCP has low liability abuse. In regards to the receptor mechanisms, our pharmacological approach (along with our transgenic approach utilizing with CB1, CB2, GPR55 knockout mice) indicates that BCP produces attenuating effects on cocaine self‐administration that are mediated by non‐cannabinoid receptors. We are now exploring the involvement of peroxisome proliferator‐activated receptor (PPAR) and transient receptor potential vanilloid 1 cation channels (TRPV1) in BCP action against cocaine. Our findings suggest that BCP shows exceptional promise as a therapeutic candidate in the treatment of cocaine use disorder. Importantly, given its good oral bioavailability and the advantage of being an already FDA‐approved non‐toxic dietary additive, BCP is a valuable candidate for drug repurposing programs in translational medicine.

Protective Effect of Beta-Caryophyllene on Doxorubicin Induced Multiple Organ Toxicity in Rats

The major limiting factor in doxorubicin’s long-term administration is the development of cumulative dosedependent cardiomyopathy and congestive heart failure. Also, doxorubicin causes deterioration in hepato-renal function. It significantly increases the levels of blood urea nitrogen, creatinine, alanine transaminase, and aspartate transaminase distortion in normal renal and hepatic histology. The present study was undertaken to find out the protective role of beta-caryophyllene (BCP), an anti-oxidant against doxorubicin-induced multiple organ toxicities in experimental animals. In this study, male Wistar rats were divided into four groups. The first group, control group, was administered with vehicle (2.5% tween 20); the second group received doxorubicin (15 mg/kg intraperitoneally at a single dose), third and fourth groups (treatment groups) received BCP plus doxorubicin (15 mg/kg) at doses of 100 mg/kg and 200 mg/kg respectively. BCP was given orally for 15 days and doxorubicin was given on 13th day of treatment. Cardiac function was assessed by measuring electrocardiogram changes and cardiac biomarkers—doxorubicin-induced significant lengthening of QT-interval and ST-elevation, which was completely prevented by BCP treatment. Doxorubicin caused oxidative stress as indicated by a significant decrease in reduced superoxide dismutase, glutathione level, and catalase activity with an increase in malondialdehyde compared to control. Doxorubicin and BCP significantly reversed these values compared to doxorubicin in heart, kidney, and liver. The histopathological examination has also shown signs of toxicity in doxorubicin treated groups, and healing effect was noticed in treatment groups.

Insulinotropic and antidiabetic effects of β-caryophyllene with l-arginine in type 2 diabetic rats.

Beta-caryophyllene (BCP) is a flavoring agent, whereas l-arginine (LA) is used as a food supplement. They possess insulinotropic and β cell regeneration activities, respectively. We assessed the antidiabetic potential of BCP, LA, and its combination in RIN-5F cell lines and diabetic rats. Ex vivo studies were carried out for glucose uptake and absorption of the combination of BCP with LA. The results indicated that the combination of BCP with LA showed a significant decrease in glucose absorption and an increase in its uptake in tissues and also an increase in insulin secretion in RIN-5F cells. The combination treatment of BCP with LA showed a significant reduction in glucose, lipid levels, and oxidative stress in pancreatic tissue when compared with the diabetic group. Furthermore, the combination of BCP with LA normalized glucose tolerance and pancreatic cell damage in diabetic rats. In conclusion, the combinational treatment showed significant potentials in the treatment of type 2 diabetes mellitus. PRACTICAL APPLICATIONS: Type 2 diabetes mellitus is the most prevalent chronic metabolic disorder affecting a large population. Beta-caryophyllene is a CB2 receptor agonist shown to have insulinotropic activity. l-Arginine is a food supplement that possesses beta-cell regeneration property. The combination of BCP with LA could work as a potential therapeutic intervention, considering the individual pharmacological activities of each. We evaluated the antidiabetic activity of the combination of BCP with LA in diabetic rats using ex vivo and in vitro experimentations. Results from the study revealed that the combination of BCP with LA showed a significant (p<.001) reduction in glucose and lipid levels as compared to individual treatment. In vitro study also supports the diabetic potential of the combination of BCP with LA in the glucose-induced insulin secretion in RIN-5F cell lines. The study indicates a therapeutic approach to treat T2DM by BCP and LA combination as food and dietary supplement.

Experimental Cannabinoid 2 Receptor Activation by Phyto-Derived and Synthetic Cannabinoid Ligands in LPS-Induced Interstitial Cystitis in Mice.

Interstitial cystitis (IC) is a chronic bladder disorder with unclear etiology. The endocannabinoid system has been identified as a key regulator of immune function, with experimental evidence for the involvement of cannabinoid receptors in bladder inflammation. This study used intravital microscopy (IVM) and behavioral testing in lipopolysaccharide-induced IC, to investigate the anti-inflammatory analgesic effects of a natural dietary sesquiterpenoid, beta-caryophyllene (BCP), which is present in cannabis among other plants, and has reported agonist actions at the cannabinoid 2 receptor (CB2R). BCP’s anti-inflammatory actions were compared to the synthetic CB2R-selective cannabinoid, HU308, and to an FDA-approved clinical treatment (dimethyl sulfoxide: DMSO). IVM data revealed that intravesical instillation of BCP and/or HU308 significantly reduces the number of adhering leukocytes in submucosal bladder venules and improves bladder capillary perfusion. The effects of BCP were found to be comparable to that of the selective CB2R synthetic cannabinoid, HU308, and superior to intravesical DMSO treatment. Oral treatment with BCP was also able to reduce bladder inflammation and significantly reduced mechanical allodynia in experimental IC. Based on our findings, we believe that CB2R activation may represent a viable therapeutic target for IC, and that drugs that activate CB2R, such as the generally regarded as safe (GRAS) dietary sesquiterpenoid, BCP, may serve as an adjunct and/or alternative treatment option for alleviating symptoms of inflammation and pain in the management of IC.

β-Caryophyllene promotes oxidative stress and apoptosis in KB cells through activation of mitochondrial-mediated pathway – An in-vitro and in-silico study

Beta-caryophyllene (BCP), are natural bicyclic sesquiterpenes which are present in numerous plants worldwide. BCP has antioxidant, antimicrobial, and antifungal properties. Here, we studied its anticancer, anti-inflammatory, and cytotoxic effects. Cells treated with BCP, in a dose-dependent manner, exhibited morphological changes, showed lower cell growth, underwent apoptosis and lost the ability to metastasis through the suppression of NF-ҡ B via PI3K/AKT signalling pathway. These results elucidate that the inhibition of NF-ҡ B and PI3K/AKT is one of the most important mechanism by which BCP suppresses cancer cell proliferation and enhances apoptosis.