Effect of Withania somnifera root extract on haloperidol-induced catalepsy in albino mice

The objective of the study was to evaluate the anticataleptic effect of Withania somnifera (WS) extract, on haloperidol-induced catalepsy in albino mice. Catalepsy was induced with haloperidol (1 mg/kg) i.p. in five groups of male albino mice (n = 6). Three groups received Withania somnifera extract (1.7, 4.25, 8.5 mg/kg) respectively, one group received scopolamine (1 mg/kg) and one group received the vehicle (1% gum acacia) orally, 30 min prior to haloperidol administration. Catalepsy was measured by using standard bar test at 30, 60, 90, 120 and 240 min. This constituted the acute study. For the chronic study, the drugs were administered for 6 more days. Catalepsy was again measured on day 7. Animals were then sacrificed by cervical dislocation and superoxide dismutase (SOD) activity was estimated in the brain. In this study, Withania somnifera extract treated groups showed a dose dependent reduction in cataleptic scores, both in the acute and chronic study. The SOD activity in brain was also found to be lowered in the WS (4.25 mg, 8.5 mg/kg) treated groups. In conclusion, Withania somnifera was found to be more efficacious than scopolamine in reversing haloperidol induced catalepsy. A clear correlation between the SOD levels and cataleptic scores was observed. We believe that the antioxidant properties of this drug could have contributed to the anticataleptic effect.     

Anti-stress activity of sitoindosides VII and VIII,new acylsterylglucosides from Withania somnifera

Two new acylsterylglucosides, sitoindoside VII and sitoindoside VIII, were isolated from the roots of Withania somnifera Dun., and were screened for putative anti-stress activity because the plant is widely regarded as the ‘Indian Ginseng’ by practitioners of the traditional Indian system of medicine. Since an acceptable paradigm of pharmacological tests for anti-stress screening has yet to be evolved, a battery of tests were employed to delineate the activity of the test compounds. The total MeOH-H₂O (1:1) extractives of the roots of W. somnifera (SG-1) and equimolecular combination of sitoindosides VII, VIII and withaferin-A, a common withanolide, (SG-2), exhibited significant anti-stress activity in all the test parameters used. The two sitoindosides also produced per se anti-stress activity, which was potentiated by withaferin-A. A preliminary acute toxicity study indicated that the compounds have a low order of acute toxicity. The anti-stress activity of SG-1 and SG-2 is consonant with the therapeutic use of W. somnifera in the Ayurveda, the Indian system of medicine.     

Tissue cultures of Withania somnifera: Morphogenesis and withanolide synthesis

Tissue cultures were established from axillary meristems of the plant Withania somnifera. Growth hormones influenced the morphogenetic responses of the cultures. The explants gave multiple shoots in Murashige and Skoog's (MS) medium supplemented with benzyladenine (BA), rooting in MS+coconut milk (CM) (10%) and callus formation in MS+2,4-dichlorophenoxyacetic acid (2,4-D) (2.0 ppm). Callus cultures failed to synthesize withanolides. Multiple shoot cultures synthesized significant levels of withanolides and their concentrations varied with different growth hormones.     

Protective effect of Withania somnifera root extract on electrographic activity in a lithium-pilocarpine model of status epilepticus

The present study investigated the anticonvulsant profile of Withania somnifera (W.s) in a lithium-pilocarpine model of status epilepticus (SE) in rats. Acute treatment with the root extract of W.s prolonged the latency to forelimb clonus but failed to protect against mortality. Acute pretreatment with W.s root extract enhanced the antiepileptic effect of diazepam and clonazepam. Rats chronically administered W.s (100, 200 mg/kg, p.o. × 7 d), when subjected to lithium-pilocarpine challenge showed a reduced mortality rate. Electrophysiological data further support the behavioural findings, as the root extract brought about a parallel change in seizure activity as paroxysmal spike activity appeared only from the 60 min record. Moreover, the seizure activity seemed to subside by 4 h in comparison with the control. The protective effect of the root extract appears to involve GABAergic mediation. © 1998 John Wiley & Sons, Ltd.     

Anticonvulsant Action of Withania somnifera (Aswaganda) Root Extract against Pentylenetetrazol-induced Kindling in Mice

Repeated administration of pentylenetetrazol (PTZ, 30 mg/kg, thrice a week for 9 weeks) produced chemical kindling when mice were challenged with a subconvulsive dose of the convulsant. These animals were also found to be susceptible to seizures after a subconvulsant dose of a beta-carboline, (FG 7142, 20 mg/kg). Withania somnifera root extract (100 mg/kg) offered a significant protection against PTZ-induced chemical kindling, and the effect was comparable to diazepam (1 mg/kg). The protective effect of W. womnifera appears to involve GABAergic mediation.     

The methanolic extract of Withania somnifera ACTS on GABAA receptors in gonadotropin releasing hormone (GnRH) neurons in mice

The effect of the methanol extract of Withania somnifera (mWS) on the gonadotropin releasing hormone (GnRH) neuron was examined in juvenile mice using the whole cell patch clamp technique. GnRH neurons are the fundamental regulators of the pulsatile release of GnRH needed for puberty and fertility. GnRH neurons were depolarized by bath application of the mWS (400 ng/μl) under the condition of a high Cl^? pipette solution in current clamp mode. In voltage clamp mode, mWS induced reproducible inward currents (31.7 ± 5.51 pA, n = 14). The mWS‐induced inward currents persisted in the presence of tetrodotoxin (TTX, 0.5 μM), but were suppressed by bicuculline methiodide (BMI, 20 μM), a GABA_A receptor antagonist. These results show that mWS affects the neuronal activities by mediating the GABA_A receptor, which suggests that WS contains an ingredient with possible GABAmimetic activity. Copyright © 2009 John Wiley & Sons, Ltd.     

Withania somnifera extract. Its effect on arterial blood pressure in anaesthetized dogs

The effect of Withania somnifera extract on arterial blood pressure in ‘normotensive’ pentobarbital anaesthetized dogs was studied. Also a possible effect of Withania somnifera on blood pressure in dogs administered either with adrenaline and acetylcholine was investigated. Thirty mongrel dogs of both sexes were distributed randomly in three series of ten animals each. Each animal was adminstered at intervals of 4 min with either a neurotransmitter, saline or the extract until a cycle of 32 minutes was completed. It is concluded that the Withania somnifera extract induced a significant decrease (p < 0.05) in the arterial and diastolic blood pressure in ‘normotensive’ pentobarbital anaesthetized dogs. Withania also significantly prevented the hypotensive effect of acetyltholine and increased the hypertensive effect of adrenaline.     

Effect of Withania somnifera root extract on reserpine-induced orofacial dyskinesia and cognitive dysfunction

Tardive dyskinesia is one of the major side effects of long-term neuroleptic treatment. The pathophysiology of this disabling and commonly irreversible movement disorder is still obscure. Vacuous chewing movements in rats are widely accepted as an animal model of tardive dyskinesia. Oxidative stress and products of lipid peroxidation are implicated in the pathophysiology of tardive dyskinesia. Repeated treatment with reserpine (1.0 mg/kg) on alternate days for a period of 5 days (days 1, 3 and 5) significantly induced vacuous chewing movements and tongue protrusions in rats. Chronic treatment with Withania somnifera root extract (Ws) for a period of 4 weeks to reserpine treated animals significantly and dose dependently (50 and 100 mg/kg) reduced the reserpine-induced vacuous chewing movements and tongue protrusions. Reserpine treated animals also showed poor retention of memory in the elevated plus maze task paradigm. Chronic Ws administration significantly reversed reserpine-induced retention deficits. Biochemical analysis revealed that chronic reserpine treatment significantly induced lipid peroxidation and decreased the glutathione (GSH) levels in the brains of rats. Chronic reserpine treated rats showed decreased levels of antioxidant defense enzymes, superoxide dismutase (SOD) and catalase. Chronic administration of Ws root extract dose dependently (50 and 100 mg/kg) and significantly reduced the lipid peroxidation and restored the decreased glutathione levels by chronic reserpine treatment. It also significantly reversed the reserpine-induced decrease in brain SOD and catalase levels in rats. The major findings of the present study indicate that oxidative stress might play an important role in the pathophysiology of reserpine-induced abnormal oral movements. In conclusion, Withania somnifera root extract could be a useful drug for the treatment of drug-induced dyskinesia.     

Transformation of Withania somnifera (L) Dunal by Agrobacterium rhizogenes: Infectivity and phytochemical studies

Three different strains of Agrobacterium rhizogenes, namely A₄, LBA 9402 and LBA 9360 were used for infection to induce hairy root formation in W. somnifera. Strain specificity of the bacterium and frequency of transformation events were analysed. Significant differences were observed between the transformation ability of the different strains of Agrobacterium. The best response in terms of transformation ability and growth of the hairy roots was obtained with A. rhizogenes strain A₄, followed by LBA 9402, whereas LBA 9360 strain failed to induce a transformation event. The production of withanolides with special reference to withaferin A was studied through HPLC in the A₄ induced hairy root lines and their respective media at different growth phases (i.e. 4, 10 and 24 weeks). The presence of withaferin A in the media as well as in the hairy roots of 10-week-old cultures highlights the importance of the present study for the commercial prospects of this plant.     

Effects of glycowithanolides from Withania somnifera on morphine-induced inhibition of intestinal motility and tolerance to analgesia in mice

Effects of glycowithanolides (Ws, 10–150 mg/kg, i.p.), consisting of sitoindosides VII-X, in combination with withaferin-A, from Withania somnifera, in Swiss mice were evaluated on (i) morphine-induced inhibition of gastrointestinal tract (GIT) transit and (ii) development of tolerance to morphine-induced analgesia. Pretreatment with Ws significantly reversed the morphine (5 mg/kg, s.c.)-induced inhibition of GIT transit at all doses. Ws (100 mg/kg, i.p., o.d., for 10 days) significantly inhibited the development of tolerance to morphine-induced analgesia. Ws per se did not influence the intestinal motility nor did it produce any perceptible analgesic effect. The present and earlier findings with Ws suggest its potential in alleviating the adverse effects of morphine and the attendant immunodepression.     

Dual inhibition of COVID-19 spike glycoprotein and main protease 3CLpro by Withanone from Withania somnifera

Objective To identify the safe and effective natural inhibitors of spike glycoprotein and main protease 3CLpro using potential natural antiviral compounds which are studied under various animal models and viral cell lines. Methods First, compounds were retrieved from the PubChem database and predicted for their druggability using the MolSoft web server, and compounds having drug-like property were predicted for major adverse drug reactions like cardiotoxicity, hepatotoxicity, arrhythmia, myocardial infarction, and nephrotoxicity using ADVERpred. Docking of nontoxic antiviral compounds with spike glycoprotein and main protease 3CLpro was performed using AutoDock vina by PyRx 0.8 version. The stability of compound-protein interactions was checked by molecular dynamic (MD) simulation using Schrodinger Desmond software. Results Based on the druggable and nontoxic profile, nine compounds were selected. Among them, Withanone from Withania somnifera showed the highest binding affinity and best fit at active sites 1 of spike glycoprotein (glycosylation site) and main protease 3CLpro via interacting with active site amino acid residues before and after MD simulation at 50 ns. Withanone, which may reduce the glycosylation of SARS-CoV-2 via interacting with Asn343 and inhibit viral replication. Conclusion The current study reports Withanone as a non-toxic antiviral against SARS-CoV-2 and serve as a potential lead hit for further experimental validation.     

Effect of Withania somnifera on B16F-10 melanoma induced metastasis in mice

Withania somnifera, a plant with known immunopotentiating activity and its bioactive fraction-Withanolide D were studied for their anti-metastatic activity using B16F-10 melanoma cells in C57BL/6 mice. Simultaneous administration of Withania extract (122 +/- 10 tumour nodules) and Withanolide (126 +/- 9 lung tumour nodules) could significantly (p < 0.001) inhibit the metastatic colony formation of the melanoma in lungs. 72.58% by extract and 69.84% by Withanolide treated, as compared to the untreated control animals also increased the survival days. Lung collagen hydroxyproline content was highly elevated in the control animals (23.5 +/- 0.9 micro g/mg protein), which was reduced by the simultaneous administration of both the extract (16.3 +/- 2.0 micro g/mg protein) and Withanolide (15.3 +/- 1.8 micro g/mg protein). The level of lung hexosamines (4.85 +/- 0.20 mg/100 mg tissue) and uronic acids (330.1 +/- 23.7 micro g/100 mg tissue) content was also elevated in the control animals. The elevated level of hexosamine was significantly reduced by the treatment with extract (1.92 +/- 0.05) and Withanolide (1.85 +/- 0.05). Similarly, the uronic acid content was also been reduced by the simultaneous administration of both Withania extract (194.2 +/- 17.4) and Withanolide (183.2 +/- 8.8). The control animals had 35.3 +/- 3.8 U/L gamma-glutamyl transpeptidase (gamma-GT), which was reduced by 50% by the treatment of extract and Withanolide to 17.5 +/- 4.0 U/L and 16.3 +/- 4.4 U/L respectively. There was a significant reduction in the levels of sialic acid in the serum of Withania extract (60.7 +/- 7.7) and Withanolide (67.16 +/- 5.8) treated animals compared to the higher level (102.2 +/- 8.7) in the control animals. Histopathological analysis of the lung tissues also correlated with these findings. Prophylactic administrations of both extract as well as Withanolide were ineffective in inhibiting the metastasis of B16F-10 melanoma cells.     

The relationship between chondroprotective and antiinflammatory effects of Withania somnifera root and glucosamine sulphate on human osteoarthritic cartilage in vitro

Using a validated explant model of in vitro cartilage damage, the effects of aqueous extracts of Withania somnifera (Ashwagandha) root and glucosamine sulphate (GlcS) were tested on the levels of nitric oxide (NO) and glycosaminoglycans (GAGs) secreted by knee cartilage from chronic osteoarthritis (OA) patients. W. somnifera extracts significantly decreased NO release by explants from one subset of patients (antiinflammatory response) and significantly increased levels of NO and GAGs released by explants from the second subset ('non-responders'). This is the first study showing direct, statistically significant, antiinflammatory effects of W. somnifera on human OA cartilage. It also confirmed that glucosamine sulphate exhibited statistically significant, antiinflammatory and chondroprotective activities in human OA cartilage. However, these beneficial effects of GlcS were observed in cartilage explants from 50% of patients tested ('responders'). In contrast, glucosamine significantly increased secretion of NO but not GAGs in explants from the second subset of OA patients ('non-responders'). Cartilage explants from the 11 OA patients gave differential responses to both drugs. Patient samples which responded to the antiinflammatory effects of W. somnifera did not always give a similar response to glucosamine, and vice versa. Thus, this in vitro model of human cartilage damage provides qualitative and statistically significant, quantitative pre-clinical data on antiinflammatory and chondroprotective activities of antiarthritic drugs.     

Acute and Sub‐Acute Oral Toxicity Assessment of the Hydroalcoholic Extract of Withania somnifera Roots in Wistar Rats

Withania somnifera is a widely used medicinal plant for several disorders. Toxicity studies on Withania somnifera are not available. Acute and sub‐acute oral toxicities of Withania somnifera root extract in Wistar rats were evaluated in the present study. In the acute toxicity study, WSR extract was administered to five rats at 2000 mg/kg, once orally and were observed for 14 days. No toxic signs/mortality were observed. In the sub‐acute study, WSR extract was administered once daily for 28 days to rats at 500, 1000 and 2000 mg/kg, orally. No toxic signs/mortality were observed. There were no significant changes (P < 0.05) in the body weights, organ weights and haemato‐biochemical parameters in any of the dose levels. No treatment related gross/histopathological lesions were observed. The present investigation demonstrated that the no observed adverse effect level was 2000 mg/kg body weight per day of hydroalcoholic extract of W. somnifera in rats and hence may be considered as non‐toxic. Copyright © 2012 John Wiley & Sons, Ltd.     

Immunomodulatory and CNS effects of sitoindosides IX and X,two new glycowithanolides from Withania somnifera

Two new glycowithanolides, sitoindoside IX (1) and sitoindoside X (2), isolated from Withania somnifera Dun., were evaluated for their immunomodulatory and CNS effects (anti-stress, memory and learning) in laboratory animals, because the plant extract is used by practitioners of the Indian systems of medicine for similar purposes. The two compounds, in doses of 100–400 μg/mouse, produced statistically significant mobilization and activation of peritoneal macrophages, phagocytosis and increased activity of the lysosomal enzymes secreted by the activated macrophages. Both these compounds (50–200 mg/kg p.o.) also produced significant anti-stress activity in albino mice and rats and augmented learning acquisition and memory retention in both young and old rats. These findings are consistent with the use of W. somnifera, in Ayurveda, to attenuate cerebral function deficits in the geriatric population and to provide non-specific host defence.     

Neuroprotective effects of Withania somnifera Dunn. in hippocampal sub-regions of female albino rat

The neuroprotective effects of W. somnifera were studied on stressed adult female Swiss albino rats. Experimental rats were subjected to immobilization stress for 14 h and were treated with a root powder extract of W. somnifera available as Stresscom capsules (Dabur India Ltd). Control rats were maintained in completely, non stressed conditions. Thionin stained serial coronal sections (7 microm) of brain passing through the hippocampal region of stressed rats (E(1) group) demonstrated 85% degenerating cells (dark cells and pyknotic cells) in the CA(2) and CA(3) sub-areas. Treatment with W. somnifera root powder extract significantly reduced (80%) the number of degenerating cells in both the areas. The study thus demonstrates the antistress neuroprotective effects of W. somnifera.     

A systematic review of the clinical use of Withania somnifera (Ashwagandha) to ameliorate cognitive dysfunction

Many developed countries are experiencing a rapidly “greying” population, and cognitive decline is common in the elderly. There is no cure for dementia, and pharmacotherapy options to treat cognitive dysfunction provide limited symptomatic improvements. Withania somnifera (Ashwagandha), a popular herb highly valued in Ayurvedic medicine, has often been used to aid memory and cognition. This systematic review thus aimed to evaluate the clinical evidence base and investigate the potential role of W. somnifera in managing cognitive dysfunction. Using the following keywords [withania somnifera OR indian ginseng OR Ashwagandha OR winter cherry] AND [brain OR cognit* OR mental OR dementia OR memory], a comprehensive search of PubMed, EMBASE, Medline, PsycINFO and Clinicaltrials.gov databases found five clinical studies that met the study's eligibility criteria. Overall, there is some early clinical evidence, in the form of randomized, placebo‐controlled, double‐blind trials, to support the cognitive benefits of W. somnifera supplementation. However, a rather heterogeneous study population was sampled, including older adults with mild cognitive impairment and adults with schizophrenia, schizoaffective disorder, or bipolar disorder. In most instances, W. somnifera extract improved performance on cognitive tasks, executive function, attention, and reaction time. It also appears to be well tolerated, with good adherence and minimal side effects.     

In vivo anti-tussive activity and structural features of a polysaccharide fraction from water extracted Withania somnifera

Aim of the study

Anti-tussive drugs are amongst the most widely used medications worldwide; however no new class of drugs has been introduced into the market for many years. The present study aims at evaluating the structural features and in vivo anti-tussive activity of a polysaccharide fraction from water extracted Withania somnifera.

Materials and methods

Herein, we have analyzed water extracted material of Withania somnifera using chemical, chromatographic, spectroscopic and biological methods.

Results

A polysaccharide fraction (F3) containing arabinosyl, galactosyl and galacturonosyl residues were obtained by anion exchange chromatography of the water extracted material. This polymer is branched and contained (1,5)-/(1,3,5)-linked arabinofuranosyl, (1,3)-/(1,6)-/(1,3,6)-linked galactopyranosyl residues together with small amount of terminal rhamnopyranosyl and terminal arabinofuranosyl residues. Peroral administration of this pectic arabinogalactan in a dose of 50 mg kg⁻¹ body weight (b.w.) decreased the number of cough efforts induced by citric acid in guinea pigs like that of codeine.

Conclusions

This study provides a scientific basis for the past and present ethnomedical uses of this plant.     

Adaptogenic activity of a novel, withanolide-free aqueous fraction from the roots of Withania somnifera Dun.

The practitioners of the traditional Indian system of medicine regard Withania somnifera Dun. as the 'Indian ginseng'. A new withanolide-free aqueous fraction was isolated from the roots of this plant and was evaluated for putative antistress activity against a battery of tests such as hypoxia time, antifatigue effect, swimming performance time, swimming induced gastric ulceration and hypothermia, immobilization induced gastric ulceration, autoanalgesia and biochemical changes in the adrenal glands. This bioactive fraction exhibited significant antistress activity in a dose-related manner in all the parameters studied. The extract of Withania somnifera root (a commercial preparation available locally) was used to compare the results. A preliminary acute toxicity study in mice showed a good margin of safety.     

Effects of glycowithanolides from Withania somnifera on an animal model of Alzheimer's disease and perturbed central cholinergic markers of cognition in rats

The active principles of Withania somnifera (WS, 20–50 mg/kg, p.o.), consisting of equimolar amounts of sitoindosides. VII–X and withaferin A, were investigated for putative nootropic activity in an experimentally validated Alzheimer's disease (AD) model. The syndrome was induced by ibotenic acid (IA) lesioning of the nucleus basalis magnocellularis (NBM) in rats. Cognitive deficits induced in NMB-lesioned rats were assessed by attenuation of a learned active avoidance task and a decrease in frontal cortical and hippocampal acetylcholine (ACh) concentrations, choline acetyltransferase (ChAT) activity and muscarinic cholinergic receptor (MCR) binding. IA-induced NBM lesioning in rats caused a marked cognitive deficit, as evidenced by severe reduction of the learned task, and was accompanied by a significant decrease in frontal cortex and hippocampal ACh levels, ChAT activity and MCR binding. WS (50 mg/kg) significantly reversed both IA-induced cognitive deficit and the reduction in cholinergic markers after 2 weeks of treatment. The findings validate the medharasayan (promoter of learning and memory) effect of W. somnifera, as has been reported in Ayurveda.