Withania Somnifera Root Extract Research Articles

Viwithan, a Standardized Withania somnifera Root Extract Induces Apoptosis in Murine Melanoma Cells.

Background:Withania somnifera is an Indian medicinal herb known for the multipotential ability to cure various therapeutic ailments as described in the ayurvedic system of medicine.Objective:In the present study, we have evaluated the antiproliferative activity of a standardized W. somnifera root extract (Viwithan) against different human and murine cancer cell lines.Materials and Methods:The cytotoxicity of Viwithan was determined using thiazolyl blue tetrazolium blue assay and crystal violet staining. The apoptotic changes in B16F1 cells following treatment with Viwithan were observed by acridine orange/ethidium bromide (AO/EB) staining and DNA fragmentation assay. The binding affinity of withanolides in Viwithan with antiapoptotic proteins B-cell lymphoma 2, B-cell lymphoma-extra large, and myeloid cell leukemia 1 (MCL-1) were studied using in silico approach.Results:The half maximal inhibitory concentration (IC50) values of Viwithan against liver hepatocellular carcinoma, Henrietta Lacks cervical carcinoma cells, human colorectal carcinoma cell line, and Ehrlich ascites carcinoma cells were 1830, 968, 2715, and 633 μg/ml, respectively. Interestingly, Viwithan was highly effective against B16F1 cells with an IC50 value of 220 μg/ml after 24 h treatment. The morphological alterations of apoptotic cell death were clearly observed in the AO/EB-stained cells after treatment with Viwithan. Viwithan induced late apoptotic changes in treated B16F1 cells as evident by the ladder formation of fragmented DNA in a time-dependent manner. The findings of molecular docking showed that withanolides present in Viwithan have a more binding affinity with the antiapoptotic proteins, particularly MCL-1.Conclusion:We have reported for the first time that Viwithan with 5% withanolides has a potent cytotoxic effect, particularly against B16F1 murine melanoma cells among the different cancer cell lines tested.SUMMARY The present study reports for the first time that Viwithan, a standardized 5% Withania somnifera root extract, has potent cytotoxicity against B16F1 murine melanoma cellsWe have investigated the in vitro cytotoxicity of Viwithan in different human and murine cancer cells. Interestingly, we found that Viwithan was particularly very effective against B16F1 melanoma cells with a half maximal inhibitory concentration value of 220 μg/mlThe microscopic observations following acridine orange/ethidium bromide staining and DNA fragmentation assays clearly indicated that Viwithan might initiate late apoptosis in B16F1 cellsThe binding affinity of withanolides in Viwithan with antiapoptotic proteins of B-cell lymphoma 2 family was predicted using AutoDock tool. The results from in silico studies indicated a plausible synergistic effect of withanolides attributing to the Viwithan-induced apoptosis through suppression of intrinsic pathway for carcinogenesis. Abbreviations used: MTT: Thiazolyl blue tetrazolium blue; DMSO: Dimethyl sulfoxide; BSA: Bovine serum albumin; DMEM: Dulbecco's minimum essential medium; NCCS: National Centre for Cell Science; PBS: Phosphate-Buffered Saline; HepG2: Liver hepatocellular carcinoma; HeLa: Henrietta Lacks cervical carcinoma cells; HCT-116: Human colorectal carcinoma cell line; EAC: Ehrlich ascites carcinoma cells; IC50: Half maximal inhibitory concentration; AO/EB: Acridine orange/Ethidium bromide; BCL-2: B-cell lymphoma 2; BCL-XL: B-cell lymphoma-extra large; MCL-1: Myeloid cell leukemia 1; PDB: Protein Data Bank; ANOVA: Analysis of variance.

Effects of the Energy of Consciousness Healing Treatment on Physical, Spectroscopic, Thermal and Behavioral Properties of Ashwagandha Root Extract

Withania somnifera (Ashwagandha) root extract contains lot of biologically active metabolites, which have a broad range of pharmacological activities. The current study was designed to evaluate the effects of The Trivedi Effect® - Energy of Consciousness Healing Treatment (Biofield Energy Healing Treatment) on the physical, spectroscopic, thermal and behavioral properties of ashwagandha root extract using PXRD, PSD, FT-IR, UV-vis spectroscopy, TGA, and DSC analysis. Ashwagandha root extract was divided into two parts – one part was control without any Biofield Energy Treatment and another part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as The Trivedi Effect® treated sample. The PXRD analysis exhibited that both the treated and control samples were amorphous in nature. The particle size values at d10, d50, and d90 of the treated sample were decreased by 8.41%, 0.51%, and 7.88%, respectively compared with the control sample. The surface area analysis revealed that the surface area of the treated sample was significantly increased by 5% compared to the control sample. The FT-IR analysis indicated the alteration of the force constant for the functional groups of the treated sample in comparison to the control sample. The UV-vis analysis revealed that the wavelength for the maximum absorbance of the control and treated samples were at 205.3 and 205.0 nm, respectively. The TGA analysis revealed that the total weight loss was decreased by 0.65% in the treated sample compared with the control sample. The DSC analysis indicated that the onset, peak, and endset vaporization temperature of the treated sample were significantly increased by 41.97%, 23.51%, and 8.82%, respectively as compared to the control sample. The latent heat of vaporization (∆H) of the treated (239.96 J/g) sample was significantly increased by 39.84% compared with the control (171.60 J/g) sample. This indicated that the treated sample was thermally more stable as compared to the control sample. The current findings suggested that The Trivedi Effect® - Energy of Consciousness Healing Treatment (Biofield Energy Healing) might have the astounding capacity to enhance the solubility, absorption, dissolution, and bioavailability of ashwagandha root extract in various form of pharmaceutical and nutraceutical formulation by modifying its particle size and surface area. Thus, the treated ashwagandha root extract might provide better therapeutic response against against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections.

Chromatographic and Spectroscopic Characterization of the Consciousness Energy Healing Treated <i>Withania somnifera</i> (Ashwagandha) Root Extract

Withania somnifera (ashwagandha) root extract is a popular health supplement, with purported health benefits including prevention and treatment of various diseases, i.e. anxiety, stress, etc. The objective of this experiment was to evaluate the impact of Energy of Consciousness Healing Treatment (The Trivedi Effect®) on the characteristic properties of the phytoconstituents present in the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while another part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The liquid chromatographic data of the control and Biofield Energy Treated samples revealed that the retention time of the 25 chromatographic peaks remained same, whereas the peak area% i.e. the relative amount of the phytoconstituents was altered significantly. The peak area% of the treated ashwagandha root extract representing the phytoconstituents was significantly decreased in the range of 6.02% to 39.74% at Rt of 5.2, 5.3, 5.4, 5.6, 5.7, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, and 8.5 minutes compared to the control sample. On the contrary, the peak area% of the other phytoconstituents present in the treated sample was significantly increased in the range of 4.12% to 82.32% at Rt of 5.1, 6.4, 6.6, 6.8, 8.1, 8.2, 8.4, 8.6, 8.8, 8.9, 9.0, 9.1, and 9.2 minutes, respectively compared to the control sample. A total of 16 withanolides such as sitoindoside IX, viscosa lactone B, dihydrowithanolide D, withanolide A, withaferin A, ixocarpalactone A, withanolide sulfoxide, etc. were proposed from the molecular mass at m/z 605, 489, 473, 471, 505, and 992 at the retention times of 6.9, 7.1, 7.8, 8.2, 8.5, and 9.2 minutes with the help of LC-MS, GC-MS and NMR data of both the samples. Subsequently, the mass peak intensities of the treated sample were significantly changed in the range of -61.24% to 106.61% compared with the control sample at the same retention time. These findings suggest that Energy of Consciousness Healing Treatment could be advantageous for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of ashwagandha extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, arthritis, stress, cancer, diabetes, sexual disorders, aging, and other chronic infections.

Evaluation of the Physico-chemical, Thermal and Behavioral Properties of Ashwagandha Root Extract: Effects of Consciousness Energy Healing Treatment

Withania somnifera (Ashwagandha) root extract has a broad range of pharmacological activities due to the presence of many biologically active metabolites. The current study was designed to evaluate the influence of The Trivedi Effect® - Consciousness Energy Healing Treatment (Biofield Energy Healing) on the physical, spectroscopic, thermal and behavioral properties of ashwagandha root extract using PXRD, PSD, FT-IR, UV-vis spectroscopy, TGA, and DSC analysis. Ashwagandha root extract was divided into two parts – one part was control without any Biofield Energy Treatment and another part was treated with the Consciousness Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as Biofield Energy Treated sample. The PXRD analysis exhibited that both the control and treated samples were amorphous in nature. The particle size values at d10, d50, and d90 of the treated sample were increased significantly by 8.69%, 7.32%, and 6.50%, respectively compared with the control sample. Similarly, the surface area of the treated sample was significantly decreased by 12.69% compared to the control sample. The FT-IR analysis indicated no significant alteration of the force constant for the functional groups in the treated sample compared to the control sample. The UV-vis analysis revealed that the wavelength for the maximum absorbance of the control and treated samples were at 207.5 and 205.2 nm, respectively. The TGA revealed the two major thermal degradation steps and the total weight loss was decreased by 0.86% in the treated sample compared with the control sample. Consequently, the maximum thermal degradation temperature at 273.53°C and 389.39°C for two broad peaks in treated sample was increased by 0.78% and 0.31%, respectively compared to the control the sample (271.40°C and 388.48°C). The DSC analysis indicated that the evaporation temperature was significantly increased by 10.95% in the Biofield Energy Treated sample with a 5.34% reduced the latent heat of vaporization compared with the control sample. The overall thermal analysis indicated that the treated sample was thermally more stable as compared to the control sample. The Trivedi Effect® - Energy of Consciousness Healing Treatment might lead to have better powder flowability and long-term storage stability compared with the control sample. Thus, the treated ashwagandha root extract might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, ageing, and other chronic infections.

A Comprehensive Physical, Spectroscopic, and Thermal Characterization of <i>Withania somnifera</i> (Ashwagandha) Root Extract Treated with the Energy of Consciousness (The Trivedi Effect<sup>®</sup>)

Withania somnifera (Ashwagandha) root extract possesses a broad range of pharmacological activities. The aim of current study was to explore the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment (Biofield Energy Healing Treatment) on the physical, spectroscopic, and thermal properties of ashwagandha root extract using PXRD, PSD, FT-IR, UV-vis spectroscopy, TGA, and DSC analysis. Ashwagandha root extract was divided into two parts. One part was denoted as the control, while the other part was defined as The Trivedi Effect® - Biofield Energy Treated sample, which received The Trivedi Effect® Treatment remotely from eighteen renowned Biofield Energy Healers. The PXRD analysis exhibited that both the control and The Trivedi Effect® - Biofield Energy Treated samples were amorphous in nature. The particle size values at d10 and d50 of the treated sample were decreased by 4.66% and 6.02% and at d90 slightly increased by 0.99% compared with the control sample. The average surface area of the treated (0.130 m2/g) sample was increased by 4.84% compared with the control (0.124 m2/g) sample. The FT-IR results suggested that the force constant of O-H (stretching), and C-O (stretching) functional groups of the treated sample were reduced compared with the control sample. The UV-vis analysis revealed that the wavelength for the maximum absorbance of the control and treated samples were at 205.4 nm and 205.6 nm, respectively. The TGA thermograms exhibited three steps of the degradation process, and the total weight loss of the treated (86.45%) sample was increased by 1.44% compared with the control (85.22%) sample. The DSC analysis revealed that melting point of the treated (201.67°C) sample was reduced by 0.87% compared with the control (203.43°C) sample. The latent heat of fusion of the treated (6.77 J/g) sample was significantly decreased by 21.82% compared to the control (8.67 J/g) sample. Hence, the treated sample would be more advantageous for the better therapeutic responses compared to the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment might have the astounding capacity to enhance the solubility, dissolution, absorption, and bioavailability of ashwagandha root extract in various forms of pharmaceutical/nutraceutical formulations by modifying its particle size and surface area. Thus, The Trivedi Effect® - Energy of Consciousness Healing Treated ashwagandha root extract might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections.

Analgesic activity of a Withania somnifera extract in stressed mice

Withania somnifera is an important Ayurvedic Rasayana herb, the roots of which are often used in traditionally known systems of medicine as tonic or for rejuvenation purposes. Effects of a single and ten daily oral doses (10, 20 and 40 mg/kg) of an analytically well standardized Withania somnifera root extract against foot shock stress triggered transient hyperthermia and hotplate test for analgesics in male mice were quantified. Body weights and basal rectal temperatures of animals were recorded on all observational days and on the 11th and 12th day of the experiment, all animals were subjected to tail suspension and pentobarbital hypnosis tests respectively. Daily dose dependent efficacy of the tested extract in stress induced hyperthermia test and in hot plate test increased with increasing number of treatment days, and its dose dependent inhibitory effect on immobility time in tail suspension were observed after its 11 daily oral doses. Daily handling and intermittent foot shock stress triggered body weight losses and elevations in basal core temperatures were almost completely prevented even by its lowest daily dose (10 mg/kg/day) tested. Repeated daily low oral doses of the Withania somnifera extract is effective in suppressing diverse stress responses, and its centrally acting analgesics and anxiolytics or antidepressants like efficacies increase with increasing numbers of treatment days. These observations reaffirm that Withania somnifera is an adaptogenic herb, and suggest that its effective therapeutic doses has to be adjusted according to the pre-existing allostatic load of patients.

Withaferin A induces Nrf2-dependent protection against liver injury: Role of Keap1-independent mechanisms

Small molecules of plant origin offer presumptively safe opportunities to prevent carcinogenesis, mutagenesis and other forms of toxicity in humans. However, the mechanisms of action of such plant-based agents remain largely unknown. In recent years the stress responsive transcription factor Nrf2 has been validated as a target for disease chemoprevention. Withania somnifera (WS) is a herb used in Ayurveda (an ancient form of medicine in South Asia). In the recent past, withanolides isolated from WS, such as Withaferin A (WA) have been demonstrated to be preventive and therapeutic against multiple diseases in experimental models. The goals of this study are to evaluate withanolides such as WA as well as Withania somnifera root extract as inducers of Nrf2 signaling, to probe the underlying signaling mechanism of WA and to determine whether prevention of acetaminophen (APAP)-induced hepatic toxicity in mice by WA occurs in an Nrf2-dependent manner. We observed that WA profoundly protects wild-type mice but not Nrf2-disrupted mice against APAP hepatotoxicity. WA is a potent inducer of Nrf2-dependent cytoprotective enzyme expression both in vivo and in vitro. Unexpectedly, WA induces Nrf2 signaling at least in part, in a Keap1-independent, Pten/Pi3k/Akt-dependent manner in comparison to prototypical Nrf2 inducers, sulforaphane and CDDO-Im. The identification of WA as an Nrf2 inducer that can signal through a non-canonical, Keap1-independent pathway provides an opportunity to evaluate the role of other regulatory partners of Nrf2 in the dietary and pharmacological induction of Nrf2-mediated cytoprotection.

Gastro-protective and Anti-stress Efficacies of Monomethyl Fumarate and a Fumaria indica Extract in Chronically Stressed Rats.

Results of the very first experiments conducted to evaluate therapeutic potentials of a fumarate containing Fumaria indica extract and of fairly low daily oral doses of monomethyl fumarate for prevention of chronic unavoidable foot-shock stress-induced gastric ulcers, and possible involvement of diverse neuro-hormonal and oxidative process in their stress response desensitizing effects are reported and discussed in this article. Preventive effects of 21 daily oral 60, 120, and 240mg/kg doses of a standardized 50% methanolic F. indica extract (MFI) and 1.25, 2.50, and 5.00mg/kg/day of pure monomethyl fumarate (MMF) were compared in rats subjected to one hour daily unavoidable foot-shocks. A pharmaceutically well-standardized Withania somnifera (WS) root extract was used as a reference herbal anti-stress agent in all experiments. Effects of the treatments on stress-induced alterations in body weight, adrenal and spleen weights, gastric ulcer and ulcer index, weight of glandular stomach, protective mucosal glycoprotein content, cellular proliferation, oxidative stress on stomach fundus, and brain tissues of male rats were quantified. Other parameters quantified were plasma corticosterone levels, brain monoamine levels, and expressions of the cytokines TNF-α, IL-10, and IL-1β in blood and brain of stressed and treated rats. Most but not every observed stress-induced anomalies were suppressed or completely prevented by both MFI and pure MMF treatments in dose-dependent manner. Qualitatively, the observed activity profiles of both of them were similar to those of WS dose tested. These results reveal that both MFI and MMF are potent gastro-protective agents against chronic unavoidable stress-induced ulcers and strongly suggest that they act as regulators or modulators of monoamine, corticosterone, and cytokine homeostasis.

Withania somnifera extract reduces the invasiveness of MDA-MB-231 breast cancer and inhibits cytokines associated with metastasis.

The aim was to examine the anti-proliferative effect of a Withania somnifera (WS) root extract in cell cultures and nude mouse xenografts of breast cancer cell line MDA-MB-231. WS root extract was used to treat tumor cells at concentrations up to 100 µg and for nude mouse experiments, the mice received daily WS at 300 mg/kg by oral gavage for 8 weeks. The WS extract reduced viability of MDA-MB-231 cells by 75% and 88% after exposure of the cells to 50 and 100 µg/mL, respectively, compared to vehicle-treated controls. WS extract caused a dose-dependent increase in the percentage of cells in the sub-G1 phase compared to untreated controls by 6% and 10% after exposure to 25 and 50 µg/mL WS extract, respectively. WS extract also inhibited proliferation of xenografted MDA-MB-231 cells. The WS extract caused reductions in xenograft size by 60% compared to the untreated control after 8 weeks of treatment. Six of ten mice in the control group showed tumor metastasis to the lung, whereas there was none in the mice treated with the WS extract. At the gene level, WS caused a 75% reduction in chemokine CCL2 expression (P < 0.05) in the xenografted tumors of the treated mice. WS root extract inhibited proliferation of breast cancer cells in vitro and in vivo and significantly reduced expression of the cytokine, CCL2. These results warrant further studies to assess the underlying molecular mechanism of the anti-tumor activity of the WS extract in breast cancer.

Withanolide A prevents neurodegeneration by modulating hippocampal glutathione biosynthesis during hypoxia.

Withania somnifera root extract has been used traditionally in ayurvedic system of medicine as a memory enhancer. Present study explores the ameliorative effect of withanolide A, a major component of withania root extract and its molecular mechanism against hypoxia induced memory impairment. Withanolide A was administered to male Sprague Dawley rats before a period of 21 days pre-exposure and during 07 days of exposure to a simulated altitude of 25,000 ft. Glutathione level and glutathione dependent free radicals scavenging enzyme system, ATP, NADPH level, γ-glutamylcysteinyl ligase (GCLC) activity and oxidative stress markers were assessed in the hippocampus. Expression of apoptotic marker caspase 3 in hippocampus was investigated by immunohistochemistry. Transcriptional alteration and expression of GCLC and Nuclear factor (erythroid-derived 2)–related factor 2 (Nrf2) were investigated by real time PCR and immunoblotting respectively. Exposure to hypobaric hypoxia decreased reduced glutathione (GSH) level and impaired reduced gluatathione dependent free radical scavenging system in hippocampus resulting in elevated oxidative stress. Supplementation of withanolide A during hypoxic exposure increased GSH level, augmented GSH dependent free radicals scavenging system and decreased the number of caspase and hoescht positive cells in hippocampus. While withanolide A reversed hypoxia mediated neurodegeneration, administration of buthionine sulfoximine along with withanolide A blunted its neuroprotective effects. Exogenous administration of corticosterone suppressed Nrf2 and GCLC expression whereas inhibition of corticosterone synthesis upregulated Nrf2 as well as GCLC. Thus present study infers that withanolide A reduces neurodegeneration by restoring hypoxia induced glutathione depletion in hippocampus. Further, Withanolide A increases glutathione biosynthesis in neuronal cells by upregulating GCLC level through Nrf2 pathway in a corticosterone dependenet manner.

Abstract 246: Mammary cancer preventive activity of an extract of withania somnifera roots

Abstract Mammary cancer preventive activity of an alcoholic extract of Withania somnifera (WS) roots was examined in female ErbB2+/- (MMTV/Neu) mice. The WS root extract was mixed with feed at a concentration of 750 mg/kg of diet. Mice treated with the root extract developed fewer mammary tumors relative to mice in the untreated group; after 310 days, mice in the treated group had an average of 1.62 tumors, and mice in the control group had 2.48, a reduction of 33% (p=0.1). Representative samples of the mammary tumors were examined histologically after staining with hematoxylin and eosin (H&amp;E) and for proliferation (Ki67 and PCNA). For mammary tumors of control mice, sections stained with H&amp;E showed high mitotic activity, whereas in tumors from treated mice, there was low mitotic activity. Labeling indices for Ki67 and PCNA markers in cancers of treated mice were 35% and 30% lower, respectively, than those of the control group. Expression of the cytokine CCL2 was 50% lower in cancers of the treated group. These results indicate that the root extract of WS reduces mammary tumor formation and tumor cell proliferation. Supported by grant U54 CA118948. Citation Format: kamel F. Khazal, Donald Hill, Clinton J. Grubbs. Mammary cancer preventive activity of an extract of withania somnifera roots. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 246. doi:10.1158/1538-7445.AM2014-246

Protective effects of Andrographis paniculata extract and pure andrographolide against chronic stress-triggered pathologies in rats.

This study was designed to experimentally verify the possibility that Andrographis paniculata could be another medicinal herb potentially useful for prevention of diverse spectrums of pathologies commonly associated with chronic unavoidable environmental stress, and whether andrographolide could as well be its quantitatively major bioactive secondary metabolite. Preventive effects of 21 daily oral 50, 100 and 200mg/kg doses of a therapeutically used extract of the plant (AP) and 30 and 60mg/kg/day of pure andrographolide were compared in rats subjected to 1-h daily unavoidable foot-shocks. A pharmaceutically well-standardized Withania somnifera (WS) root extract was used as a reference herbal anti-stress agent in all experiments. Effects of the treatments on stress-induced alterations in body weight, gastric ulcer, adrenal and spleen weights, and depressive state and sexual behavior in male rats were quantified. Other parameters quantified were plasma cortisol levels, and expressions of the cytokines TNF-α, IL-10 and IL-1β in blood and brain. All observed stress-induced pathological changes were less pronounced or completely prevented by both AP and pure andrographolide. Even the lowest tested doses of AP (50mg/kg/day) or of andrographolide (30mg/kg/day) suppressed almost maximally the blood IL-1β and IL-10 as well as brain TNF-α and IL-10 expressions induced by chronic stress. Qualitatively, the observed activity profiles of both of them were similar to those of WS dose tested. These results reveal that both AP and andrographolide are pharmacologically polyvalent anti-stress agents, and that biological processes regulating corticosterone and cytokine homeostasis are involved in their modes of actions.

Prenatal developmental toxicity evaluation of Withania somnifera root extract in Wistar rats

Context: Withania somnifera (L) Dunal (Solanaceae) is an important traditional herbal medicine used for thousands of years and is considered as the Indian ginseng. Reports on the effect of Withania somnifera root (WSR) extract on the developing foetus of pregnant rats including mortality, structural abnormalities, changes in growth and effects on dams are not available. Objective: The present study was performed to evaluate the prenatal developmental toxicity potential of WSR extract in rats. Materials and methods: WSR extract was given orally to pregnant rats during the period of major organogenesis and histogenesis (days 5 to 19 of gestation) at the dose levels of 500, 1000 and 2000 mg/kg/day. Clinical observations including mortality, moribundity, behavioural changes, signs of overt toxicity, body weight, gross pathological changes of dams and foetal analyses including external malformations, skeletal and soft tissue malformations were evaluated. Results: No evidence of maternal or foetal toxicity was observed. WSR extract caused no changes (p < 0.05) in body weight of parental females, number of corpora lutea, implantations, viable foetuses, external, skeletal and visceral malformations. Discussion and conclusion: Under the conditions of the study, the no-observed-effect level (NOEL) of WSR extract for maternal and developmental toxicity was concluded to be at least 2000 mg/kg/day.

Ashwagandha (Withania somnifera) reverses β-amyloid1-42 induced toxicity in human neuronal cells: implications in HIV-associated neurocognitive disorders (HAND).

Alzheimer’s disease (AD) is characterized by progressive dysfunction of memory and higher cognitive functions with abnormal accumulation of extracellular amyloid plaques and intracellular neurofibrillary tangles throughout cortical and limbic brain regions. At present no curative treatment is available, and research focuses on drugs for slowing disease progression or providing prophylaxis. Withania somnifera (WS) also known as ‘ashwagandha’ is used widely in Ayurvedic medicine as a nerve tonic and memory enhancer. However, there is a paucity of data on the potential neuroprotective effects of W.somnifera against β-Amyloid (1–42)-induced neuropathogenesis. In the present study, we have tested the neuroprotective effects of methanol:Chloroform (3:1) extract of ashwagandha against β-amyloid induced toxicity and HIV-1Ba-L (clade B) infection using a human neuronal SK-N-MC cell line. Our results showed that β-amyloid induced cytotoxic effects in SK-N-MC cells as shown by decreased cell growth when tested individually. Also, confocal microscopic analysis showed decreased spine density, loss of spines and decreased dendrite diameter, total dendrite and spine area in clade B infected SK-N-MC cells compared to uninfected cells. However, when ashwagandha was added to β-amyloid treated and HIV-1 infected samples, the toxic effects were neutralized. Further, the MTT cell viability assays and the peroxisome proliferator-activated receptor-γ (PPARγ) levels supported these observations indicating the neuroprotective effect of WS root extract against β-amyloid and HIV-1Ba-L (clade B) induced neuro-pathogenesis.

Myeloid-derived suppressor cell function is reduced by Withaferin A, a potent and abundant component of Withania somnifera root extract.

Myeloid cells play a crucial role in tumor progression. The most common tumor-infiltrating myeloid cells are myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAMs). These cells promote tumor growth by their inherent immune suppressive activity which is enhanced by their cross-talk. The root extract of the plant Withania somnifera (Ashwagandha) (WRE) has been reported to reduce tumor growth. HPLC analysis identified Withaferin A (WA) as the most abundant constituent of WRE and led us to determine whether the anti-tumor effects of WRE and WA involve modulating MDSC and TAM activity. A prominent effect of MDSC is their production of IL-10 which increases upon cross-talk with macrophages, thus polarizing immunity to a pro-tumor type 2 phenotype. In vitro treatment with WA decreased MDSC production of IL-10 and prevented additional MDSC production of IL-10 generated by MDSC-macrophage cross-talk. Macrophage secretion of IL-6 and TNFα, cytokines that increase MDSC accumulation and function, was also reduced by in vitro treatment with WA. Much of the T-cell suppressive activity of MDSC is due to MDSC production of reactive oxygen species (ROS), and WA significantly reduced MDSC production of ROS through a STAT3-dependent mechanism. In vivo treatment of tumor-bearing mice with WA decreased tumor weight, reduced the quantity of granulocytic MDSC, and reduced the ability of MDSC to suppress antigen-driven activation of CD4+ and CD8+ T cells. Thus, adjunctive treatment with WA reduced myeloid cell-mediated immune suppression, polarized immunity toward a tumor-rejecting type 1 phenotype, and may facilitate the development of anti-tumor immunity.

Withaferin A, a potent and abundant component of Withania somnifera root extract, reduces myeloid-derived suppressor cell function (P2103)

Abstract Myeloid cells play a crucial role in growth and metastasis of malignant tumors. Tumor infiltrating myeloid cells include myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages. These cells infiltrate into tumor and suppress tumor immunity by their inherent immune suppressive activity which is enhanced by interactions with each other (cross-talk). The root extract of the plant Withania somnifera (WRE) has been reported to reduce tumor cell proliferation and angiogenesis. We hypothesize that WRE or its constituents impact tumor infiltrating myeloid cells and thereby boost anti-tumor immunity. HPLC and MS analysis revealed that Withaferin A (WA) is the most abundant constituent of WRE. A prominent effect of MDSC is their production of IL-10 which increases upon cross-talk with macrophages, promoting type-2 immunity. WA reduces constitutive and cross-talk induced IL-10 secretion from MDSC. Macrophage secretion of IL-6 and TNFa cytokines that are characteristic of M1-type macrophages and that also increase MDSC accumulation and function, are also reduced by WA. Much of the T cell suppressive activity of MDSC is due to MDSC production of reactive oxygen species (ROS), and WA significantly reduces MDSC production of ROS. Thus adjunctive treatment with WA has the potential to concomitantly reduce myeloid cell mediated immune suppression, to polarize immunity towards a tumor-rejecting type I phenotype, and to facilitate the development of anti-tumor immunity.

Abstract 4718: Withaferin A, a potent and abundant component of Withania somnifera root extract, reduces myeloid-derived suppressor cell function.

Abstract Myeloid cells play a crucial role in growth and metastasis of malignant tumors. Tumor infiltrating myeloid cells includes myeloid-derived suppressor cells (MDSC), tumor-associated macrophages (TAMS) and dendritic cells (DC). These cells infiltrate into tumor and suppress tumor immunity by their inherent immune suppressive activity which is enhanced by interactions with each other (cross-talk). Therapeutic strategies targeting manipulation of these cells are being developed. The root extract of the plant Withania somnifera (Ashwagandha) (WRE) has been reported to reduce tumor cell proliferation and angiogenesis. Whether the anti-tumor effects of WRE are due to direct effects on malignant cells, or whether WRE also impacts immune suppressive myeloid cells is unknown. We hypothesize that WRE or its constituents impact tumor infiltrating myeloid cells and thereby boost anti-tumor immunity. HPLC and mass spectrometry analysis revealed that Withaferin A (WA) is the most abundant constituent of Withania somnifera root powder (ChromaDex, Inc. Irvine, CA). In comparison with the other constituents, WA has the most potent effect on reducing suppressive functions of myeloid cells. A prominent effect of MDSC is their production of IL-10 which increases upon cross-talk with macrophages, thus polarizing immunity to a tumor promoting type-2 phenotype. WA reduces inherent and cross-talk induced IL-10 secretion from MDSC in a dose dependent manner. Macrophage secretion of IL-6 and TNFα cytokines that are characteristic of M1-type macrophages and that also increase MDSC accumulation and function, are also reduced by WA. Much of the T cell suppressive activity of MDSC is due to MDSC production of reactive oxygen species (ROS), and WA significantly reduces MDSC production of ROS. Thus adjunctive treatment with WA has the potential to concomitantly reduce myeloid cell mediated immune suppression, to polarize immunity towards a tumor-rejecting type I phenotype, and to facilitate the development of anti-tumor immunity. Supported by NIH RO1CA115880, RO1CA84232, and American Cancer Society IRG-97-153-07 Citation Format: Pratima Sinha, Suzanne Ostrand-Rosenberg. Withaferin A, a potent and abundant component of Withania somnifera root extract, reduces myeloid-derived suppressor cell function. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4718. doi:10.1158/1538-7445.AM2013-4718

Withania somnifera ameliorates lead-induced augmentation of adrenergic response in rat portal vein

Objectives:Present study was undertaken to elucidate the ameliorating potential of Withania somnifera root extract (WRE) against lead-induced augmentation of adrenergic response in rat portal vein.Materials and Methods:In-vitro studies were conducted on effect of lead alone and lead+WRE on rat-isolated portal vein while in-vivo studies were done in three groups of 12 rats each; Group-II and III received 0.5% lead acetate and 1.0% WRE + 0.5% lead acetate, respectively, in drinking water for 12 weeks whereas group-I served as control. Adrenaline and noradrenaline levels in brain and blood were determined by HPLC assay while vascular reactivity of portal vein to lead and WRE was determined by measuring the isometric tension.Results:Following in-vitro exposure, lead did not alter the contractile effect of phenylephrine. In-vivo studies revealed that contractile effect of lead on portal vein was significantly potentiated and it was antagonized by prazosin (10-7 M) and WRE (1%). WRE treatment significantly reduced elevated blood noradrenaline (37.80%) and restored noradrenaline level in brain (39.39%) in lead-exposed animals. These values were almost comparable to the control group. But it failed to significantly affect the blood and brain adrenaline levels.Conclusions:Results suggest that following pre-exposure of rats to WRE, lead-induced augmentation of alpha1-adrenoceptors mediated response was reversed possibly by regulating catecholamine release from nerve endings. Thus, WRE may be useful in therapeutic management of lead-induced hypertension.

Antiviral Activity of Ashwagandha Extract on Infectious Bursal Disease Virus Replication

In the present study Ashwagandha plant was screened for antiviral activity against Infectious Bursal Disease Virus Replication. The hydro- alcoholic extract of Withania somnifera roots showed the inhibition of virus at maximum 99.9 % in its highest nontoxic concentration, 25μg/ml in Cytopathic Effect Reduction Assay. Results indicated that the Withania somnifera root extract has antiviral property against Infectious Bursal Disease virus.

Rectal gel application of Withania somnifera root extract expounds anti-inflammatory and muco-restorative activity in TNBS-induced Inflammatory Bowel Disease

BackgroundInflammatory Bowel Disease (IBD) is marked with chronic inflammation of intestinal epithelium driven by oxidative stress. Traditional treatments with plant extracts gained renewed interest due to their ability to ameliorate the multi factorial conditions like inflammation. We investigated the beneficial effects of Withania somnifera in Trinitro Benzyl Sulfonic Acid (TNBS) induced experimental IBD through a rectally applicable formulation.MethodsThe study included (i) preparation of gel formulation from aqueous Withania somnifera root extract (WSRE), (ii) biochemical assays to determine its performance potential, (iii) testing of formulation efficacy in TNBS-induced IBD rat model, and (iv) histo-patholgical studies to assess its healing and muco-regenerative effect in IBD-induced rats. For this purpose, concentration dependant antioxidant activity of the extracts were evaluated using biochemical assays like (a) inhibition of lipid peroxidation, (b) NO scavenging, (c) H2O2 scavenging, and (d) ferric reducing power assay.ResultsThe extract, at 500 μg/ml, the highest concentration tested, showed 95.6% inhibition of lipid peroxidation, 14.8% NO scavenging, 81.79% H2O2 scavenging and a reducing capacity of 0.80. The results were comparable with standard antioxidants, ascorbic acid and curcumin. WSRE treatment positively scored on histopathological parameters like necrosis, edema, neutrophil infiltration. The post treatment intestinal features showed restoration at par with the healthy intestine. In view of these results, gel formulation containing an aqueous extract of W. somnifera, prepared for rectal application was tested for its anti-inflammatory activity in TNBS-induced rat models for IBD. Commercially available anti-inflammatory drug Mesalamine was used as the standard in this assay.ConclusionsDose of the rectal gel applied at 1000 mg of WSRE per kg rat weight showed significant muco-restorative efficacy in the IBD-induced rats, validated by histo-pathological studies.