Phytochemical analysis of Tinospora cordifolia and Withania somnifera and their therapeutic activities with special reference to COVID-19

Various important medicines make use of secondary metabolites that are produced by plants.Medicinal plants,such as Withania somnifera and Tinospora cordifolia,are rich sources of chemically active compounds and are reported to have numerous therapeutic applications.The therapeutic use of medicinal plants is widely mentioned in Ayurveda and has folkloric importance in different parts of the world.The aim of this review is to summarize the phytochemical profiles,folkloric importance,and primary pharmacological activity of W.somnifera and T.cordifolia with emphasis on their action against the novel coronavirus.

Effect of Withania somnifera root extract on the sexual behaviour of male rats

Aim: To determine the effect of a methanolic extract of Withania somnifera (L.) Dunal roots on sexual competence of male rats. Methods: Male rats were orally administered 3000 mg'kg^1.day^1 of root extract for 7 days. Their sexual behaviour was evaluated 7 days prior to treatment, day 3 and 7 of treatment, and day 7, 14 and 30 posttreatment by pairing each male with a receptive female. Results: The root extract induced a marked impairment in libido, sexual performance, sexual vigour, and penile erectile dysfunction. These effects were partly reversible on cessation of treatment. These antimasculine effects are not due to changes in testosterone levels or toxicity but may be attributed to hyperprolactinemic, GABAergic, serotonergic or sedative activities of the extract. Conclusion: Use of W. somnifera roots may be detrimental to male sexual competence. (Asian J Androl 2002 Dec;4: 295-298)

Withania somnifera: a pre-clinical study on neuroregenerative therapy for stroke

Stroke management exerts insurmountable societal and economic burden on the patient as well as their caregivers. In the year 2010 alone, the direct and indirect costs of stroke care amounted to 36.5 billion dollars （Go et al., 2014）. Despite concentrated efforts to develop a safe, effective drug for stroke, we have not discovered one since the introduction of recombinant tissue plasminogen activator （rtPA）--the standalone FDA-approved therapy for stroke. While rtPA is highly effective, it needs to be given within 3-4.5 hours of the onset of stroke symptoms （Zivin, 2009）. This is often complicated by the delay in the commencement of treatment due to preliminary inclusion parameters that are required to be ascertained before rtPA administration.

Update on the preclinical and clinical assessment of Withania somnifera: from ancient Rasayana to modern perspectives

The Rasayana plant Withania somnifera(W.somnifera)Dunal,also known as“Ashwagandha”,has been mentioned in various classical Ayurvedic texts,such as Charaka Samhita,Sushruta Samhita,and Nighantus.This Ayurvedic drug has been referred to as a tonic that renews the body,provides physical and mental vigor in weakened states,and promotes endurance and longevity.W.somnifera possesses notable biological activity in many ailments,such as diabetes,conjunctivitis,insomnia,senile dementia,Parkinson’s disease,nervous system disorders,rheumatism,and arthritis.These pharmacological activities are due to the presence of diverse active components and their derivatives.Some lead compounds are found to be effective against anxiety and other central nervous system disorders.W.somnifera has been proven to be effective and safe for a wide range of ailments from ancient to modern times.Its reported properties represent the traditional use of W.somnifera as indicated in the literature;furthermore,W.somnifera is one of the most important prescribed drugs in Ayurveda for its multimodal effects.This current review highlights the bioactive present and provides an overview of the toxicological and pharmacological studies on W.somnifera,including preclinical and clinical studies.From its earliest utilization to its current application,W.somnifera has been recognized to be effective at clinical levels for human health and welfare.Greater attention to the safety and efficacy of W.somnifera would provide more scientific evidence,promoting global acceptance of the Ayurvedic plant.

Compound’s Pre-Screening of <i>Withania somnifera</i>, <i>Bacopa monnieri</i>and <i>Centella asiatica</i>Extracts

Spectral fluorescence signature, Gas Chromatography-Mass Spectrometry and Liquid Chromatography-Mass Spectrometry for identification of chemical and bioactive compounds were applied to study the plant extracts of Withania somnifera, Centella asiatica and Bacopa monnieri which are related to the possible treatment of mental diseases as Alzheimer, Parkinson and Depression. These plants are known for different positive phytotherapeutic effects on the human brain without negative post-, adverse or after effects to the treated individuals, and have been recommended in several medical studies. Therefore, we selected these plants for further analysis, based on the inhibition results of in vitro Amyloid Beta fibrillation tests made by previous measurements. With this study a first screening of the complex plant extract mixtures was performed, to get an initial overview about known and unknown ingredients. In all three plants, similar main compounds were identified, however in different quality and quantity. These may provide substantial information on which compound combinations might be mainly responsible for the positive effects and should be further investigated being responsible for reducing the fibrillation process of Amyloid Beta.

Evaluation of the Anti-inflammatory and Sedative Effects of Leaf Aqueous Extract of Withania somnifera （L.） Dunal （Solanaceae）in Rats and Mice

The use of medicinal plants in South Africa is cultural. Withania somnifera is one of the medicinal plants used to treat various ailments in the country. The plant species has been used by traditional medicine practitioners to treat inflammation and painful conditions like rheumatism. It is also known to be used as a sedative and hypnotic drug. Despite the claims, there is no information in literature to corroborate the therapeutic success of Withania somnifera in the treatment of inflammation and insomnia. The study, therefore, investigated the anti-inflammatory and central nervous system depressant activities of the leaf aqueous extract of the plant species in mice and rats. Fresh leaves of W. somnifera were collected from Kirstenbosch Botanical Gardens, South Africa, authenticated by a taxonomist and a voucher specimen （UWC 005） deposited in the University＇s Herbarium. Leaf aqueous extract was prepared using standard extraction methods. The carrageenan-induced rat paw oedema test was used to determine the anti-inflammatory effects while pentobarbitone-induced sleep and locomotor activity tests were used to evaluate the sedative effect of the plant species. Phytochemical qualitative analysis, acute toxicity and HPLC studies of the plant species were also carried out using standard methods. The phytochemical qualitative analysis carried out on the dried powdered leaves of W. somnifera showed the presence of saponins, tannins and triterpene steroids. Leaf aqueous extract of IV. somnifera （100-200 mg/kg IP） significantly prolonged pentobarbitone （40 mg/kg, i.p.）-induced sleep in mice in a dose dependant manner. Diazepam （0.5 mg/kg, i.p.） significantly prolonged pentobarbitone （40 mg/kg, i.p.）-induced sleep in mice. The doses of 100 and 200 mg/kg （i.p.） of the plant species and 0.5 mg/kg （i.p.） of diazepam significantly reduced the locomotor activity of mice. Leaf aqueous extract of the plant species （50-200 mg/kg, i.p.） significantly reduced the oedema produced by carrageenan （1%） in rats over 90 min period of testing. Indomethacin （20 mg/kg, i.p.） significantly reduced carrageenan （1%）-induced oedema in rats over 120 min period of testing. The LDs0 value obtained for the leaf aqueous extract of the plant species following inter-peritoneal injection was 1,600 mg/kg while that following oral administration was probably over 4,000 mg/kg. The HPLC finger-print of the aqueous extract showed distinct peaks at the following retention times 2.977, 3.594, 4.154, 4.406, 4.660 and 15.267 min. The results obtained show that leaf aqueous extract of W. somnifera has both sedative and anti-inflammatory effects.

A Comparative Investigation on Growth of Three Food Born Pathogenic Bacteria Inoculated with Withania somnifera:an Invitro Experimental Study

Background:Withania somnifera(WS)is proposed as one of the alternatives instead of the antibiotic.This study is aimed to evaluate the inhibitory potency of enzymatic extract of the fruits of the WS.Methods:As an invitro experimental study,the growth rate of Shigella dysenteriae,Salmonella typhimurium,and Escherichia coli inoculated in different concentrations(25%,12.5%,6.25%and 3.125%)of the extract were assessed.A microtitre plate method was conducted.ANOVA was applied to identify statistical differences with p-value<0.05.Results:Different concentrations of extract,in comparison with control,declined the growth rate of all tested bacteria.All concentrations inhibited the growth of S.typhimurium(p<0.05).Compared to the microorganism control,effective concentration of the extract inhibiting the growth of E.coli was 12.5%,and 6.25%,while it was 12.5%,and 6.25%for Sh.dysenteriae(p<0.05).A dose-dependent response of E.coli was observed.The antibacterial activity of the extract tested was found mainly against E.coli and Sh.dysenteriae.The most resistant microorganism compared to E.coli and Sh.dysenteriae was S.typhimurium(p<0.05).25%of the concentration of the extract showed the different inhibitory effect among three tested bacteria(p<0.05).Conclusions:The extract was labeled as an antibacterial agent against the representative of three foodborne bacteria,Invitro.The common effective concentrations of the extract(12.5,and 6.25%)is recommended for further research,as food additive,to remedy digestive ailments related to E.coli,S.typhimurium and Sh.dysenteriae.

Influence of Withania somnifera on obsessive compulsive disorder in mice

Objective:To study the influence of methanolic and aqueous extract of Withania somnifera (W.somnifera) root on the marble-burying behavior of mice a well-accepted model of obsessive compulsive behavior.Methods:Mice were divided in different groups(n=6).Fluoxetine(5,10,15 mg/kg), (10,25,50,100 mg/kg) and melhanolic extract W.somnifera(MEWS)(10,25,50,100 mg/kg) were administered i.p.30 min.prior to the assessment of marble burying behavior and locomotor activity.The control group received vehicle of the extract.Results:Administration of aqueous extracts W.somnifera(AEWS) and MEWS(50 mg/kg) successively decreaesed the marble burying behavior activity without affecting motor activity.This effect of AEWS and MEWS was comparable to standard fluoxetine,ritanserin and parachlorophenylalanine.Conclusions:W.somnifera extract is effective in treating obsessive compulsive disorder.

Inhibition of the emergence of multi drug resistant Staphylococcus aureus by Withania somnifera root extracts

Objective:To search systematically for an alternative therapy with compounds particularly from plant origin.Methods:Efficacy test of different root extracts of Withania somnifera(W. somnifera)(L) Dunal against multi drug resistant(MDR) Staphylococcus aureus(S.aureus) variants was performed following the agar well diffusion method.Evaluation of susceptibility pattern of the isolates was carried out by employing disk diffusion method using standard antibiotic disks. Results:In vitro study with W.somnifera root extracts was found to be effective against all the MDR S.aureus strains isolated from local and patient sources.Different root extracts of WS showed different degree of effectiveness against the isolates.Conclusions:The major active principles responsible for the antibacterial efficacy were mainly present in methanol(MeOH) extract and ethanol(EtOH) extracts as well as in butanol(BuOH) extract fraction.Amongst all the extracts the BuOH fraction was found to be most active against all the isolates but aqueous extract was the least active one.Finally it may be concluded that the antimicrobials from W.somnifera may raise an alternative therapy for MDR staphylococcal infections in near future.

Current Status of <i>Withania somnifera</i>(L.) Dunal: An Endangered Medicinal Plant from Himalaya

Withania somnifera is highly medicinal plant species of Pakistan floristically placed in the Western Himalayan Province, Himalaya Range of Azad Jammu and Kashmir and in the Soon valley (Salt Range) of Punjab Pakistan. It grows in high altitude of 5500 feet in the Himalayas. Its English name is Winter Cherry and its Urdu name is Asghand/Kutilal. It is the low lying shrub of about 1.50 m in height with erect branching. It grows in dry arid regions of Pakistan, India, China and Bangladesh. Its flowers are bisexual, small, axillary, greenish, solitary and few-flowered cymes. Chemical analysis of Ashwagandha showed that it has alkaloids and steroidal lactones (withanolides) as the main chemical constituent. W. somnifera has not yet been assessed for the IUCN Red List. However, in Pakistan, W. somnifera is falling under criteria A of endangered category. Asghand (W. somnifera) acquires much therapeutic potential which include sedative, narcotic, thyroid stimulation, anti-inflammatory, hypnotic, anti-stress, general tonic, diuretic, antimicrobial, and antitumor activities. There are number of threats responsible for the decrease of W. somnifera number in western Himalayan range and salt range of Punjab Pakistan which may include deforestation over exploitation and climatic changes. Conservation strategies need to be adopted for improving the population pool of W. somnifera. There is also the dire need to conserve this important medicinal plant species with some standard biotechnological approaches and conservation strategies in future.

Economics of Yield and Production of Alkaloid of <i>Withania somnifera</i>(L.) Dunal

The yield parameters and cost economics of Withania somnifera were studied using Integrated Nutrient Management (INM) treatments. Withania somnifera is cultivated in around 10,780 ha with a production of 8429 tones in India. The annual demand of this herb increased from 7028 tones (2001-2002) to 9127 tones (2004-2005). The field experiment was conducted at the research farm of Department of Plant Physiology, College of Agriculture, Jabalpur during Kharif season of 2008-2009 (1st year) and 2009-2010 (2nd year), whereas the laboratory work was carried out in Quality Laboratory, College of Agriculture, Jabalpur. The soil of experimental field was sandy loam in texture with pH 7.5, EC 0.18 dsm-1 having good drainage. Soil analysis revealed that available nitrogen was low (202.0 kg·ha-1) whereas available phosphorus (16.25 kg·ha-1) and potassium (236.0 kg·ha-1) were in the medium range. The present paper shows how to determine the economics of varying INM treatments. Cultivation of W. somnifera in India is gaining popularity among farmers;however, due to poor soil fertility, and costly chemical fertilizers and pesticides its production is not economical or profitable.

催眠睡茄Withania somnifera毛状根的诱导及睡茄素A的合成

睡茄素A是存在于催眠睡茄的根和叶中的主要次生代谢产物之一。该实验以催眠睡茄的无菌苗叶片为外植体,利用发根农杆菌C58C1诱导催眠睡茄毛状根,诱导率达30%。同时,利用HPLC测定野生催眠睡茄植株的根、茎、叶和30d液体悬浮培养的毛状根中睡茄素A的含量结果表明,毛状根中睡茄素A平均质量分数为1.588 mg·g-1,是野生植株平均含量的1.96倍;其中,毛状根系M3的睡茄素A含量最高,质量分数达到1.783 mg·g-1,是野生型催眠睡茄植株根含量的1.51倍。因此,可以培养催眠睡茄毛状根来获得睡茄素A。

Withania somnifera (Ashwagandha):a Novel Source of L-asparaginase

Different parts of plant species belonging to Solanaceae and Fabaceae families were screened for L-asparaginase enzyme （E.C.3.5.1.1.）. Among 34 plant species screened for L-asparaginase enzyme. Withania somnifera L. was identified as a potential source of the enzyme on the basis of high specific activity of the enzyme. The enzyme was purified and characterized from W. somnifera, a popular medicinal plant in South East Asia and Southern Europe. Purification was carried out by a combination of protein precipitation with ammonium sulfate as well as Sephadex-gel filtration. The purified enzyme is a homodimer, with a molecular mass of 72 ± 0.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresisand size exclusion chromatography. The enzyme has a pH optimum of 8.5 and an optimum temperature of 37 ℃. The Km value for the enzyme is 6.1 × 10^-2 mmol/L. This is the first report for L-asparaginase from W. somnifera, a traditionally used Indian medicinal plant.

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

Aim: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.Methods:WS root extract was used to treat tumor cells at concentrations up to 100μg and for nude mouse experiments,the mice received daily WS at 300 mg/kg by oral gavage for 8 weeks.Results:The WS extract reduced viability of MDA-MB-231 cells by 75%and 88%after exposure of the cells to 50 and 100μg/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 50μg/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.Conclusion:WS root extract inhibited proliferation of breast cancer cells in vitro and in vivo and signifi cantly 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.

Exploring the benefits of biochar over other organic amendments for reducing of metal toxicity in Withania somnifera

Biochar is considered as a good metal sequester and ameliorates the metal toxicity and uptake in plants.However,its effec-tiveness over other organic amendments is not well discussed.The aim of this study is to compare the biochar amendments alleviating the levels of Pb and Cd in Withania somnifera L.Dunal with other organic manure.Farmyard manure,vermicom-post,and biochar were applied in the field spiked with highly available Pb and Cd.Metal bioavailability and accumulation,plant growth and plant physiology,antioxidant enzymes and metabolite content of W.somnifera along with soil properties were evaluated in all treatments.Study indicates that the application of organic manures significantly alleviated the metal levels(33-72%)in the plant in comparison with control.The herbage yield was significantly higher(33-69%)under different organic manures as compared to the metal treatments.Secondary metabolite contents and antioxidant enzymes were higher in metal treatments in comparison with control.Organic manures were not only able to restrain the Pb and Cd in soil but also enhanced the soil microbial activities.Results indicate that biochar amendments were more promising than farmyard manure and vermicompost due to the presence of more stable carbon in biochar and more alleviation in metal uptake.In addition,net profit in the cultivation of W.somnifera was higher for biochar amendments(50%higher)compared to control.The study recommended that biochar could be a better option for commercial and safer production of W.somnifera.

Exploring phytochemicals of Withania somnifera from different vicinity for functional foods

The well-known relation between food and bioactives explores the great prospective of innovative and novel food to maintain or improve health,increasing the demand investigating advance products with medicinal effect.Withania somnifera also known as‘Ashwagandha’in India,is one of the most utilized plants in different ancient medicine systems.The present work is an effort to develop a knowledge-based concept and a quality control approach for the development of functional foods.The study took into account the standardization criteria and examined how different geographies or different field sites(of India)would influence the phytoconstituents and their distribution in the different samples of W.somnifera.Phytoconstituents level were strongly influenced by soil quality,cultivar,and growing season.An increase in phenolic content,flavonoid content,antioxidants and the quality yield,were all correlated with increased phytoconstituent concentrations.The highest hydro-methanolic extract yield(by reflux(Soxhlet),sonication(20 min)and maceration(16 h)was from the sample collected from Rajasthan(18.61%,m/m)followed by West Bengal(11.21%)and Madhya Pradesh(12.67%)respectively.The methanolic extract of Rajasthan sample(produced by the reflux(Soxhlet)extraction method)had the highest concentrations of flavonoids(65.32 mg quercetin equivalent(QE)/g)and the total phenols(60.35 mg gallic acid equivalent(GAE)/g),while Madhya Pradesh sample(distilled water reflux(Soxhlet))extract had the highest antioxidant content(74.13%).High performance thin layer chromatography(HPTLC)method was also utilized for identification of withanolides A.Chromatographic separations were carried out using V(ethyl acetate):V(toluene):V(acetic acid)=9.1:1:0.6 as the mobile phase.The gas chromatograph-mass spectrometer(GC-MS)study of hydro-methanolic Soxhlet extract of W.somnifera from 3 different regions of India was done and the total phytochemical substances identified were 132 in sample from Rajasthan,107 in sample from West Bengal,and 164 in sample from Madhya Pradesh respectively.To conclude,the present study demonstrates how,within a framework of integrated agricultural practices,yield and phytochemicals merge into a new paradigm of functional foods.

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 Pub Chem database and predicted for their druggability using the Mol Soft 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 Auto Dock vina by PyRx 0.8 version.The stability of compoundprotein 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.

金银花等6种植物提取物总黄酮含量与抗氧化性相关性研究

以金银花、核桃叶、南非醉茄、何首乌、红景天、石榴皮6种提取物为试验对象,通过硝酸铝—亚硝酸钠比色法测定了总黄酮含量;采用ABTS、DPPH与Fenton法测定了其抗氧化能力并进行比较;采用Pearson法进行相关性分析。结果表明:对DPPH自由基的清除能力顺序为金银花>核桃叶>南非醉茄>红景天>何首乌>石榴皮;对羟基自由基的清除能力顺序为金银花>核桃叶>南非醉茄>何首乌>石榴皮>红景天;对ABTS+自由基清除能力顺序为金银花>核桃叶>红景天>南非醉茄>何首乌>石榴皮。总黄酮含量对DPPH、ABTS+与羟基自由基清除能力的相关系数分别为0.819,0.848,0.791,表明总黄酮含量与清除DPPH、ABTS+自由基能力呈显著相关性,与清除羟基自由基能力相关性不显著。

催眠睡茄的离体快繁及四倍体的诱导

以催眠睡茄腋芽为外植体建立快繁体系,利用秋水仙素进行催眠睡茄四倍体诱导,并将其与二倍体进行了初步的形态学和细胞学方面的比较。结果发现:催眠睡茄二倍体(2n=2×=48)最适增殖培养基为MS+6-BA0.5mg/L+NAA0.05mg/L;生根培养基为1/2MS+IBA0.2mg/L;四倍体(2n=4×=96)诱导率因秋水仙素浓度和处理时间的不同而异,其中用0.5%的秋水仙素对丛生芽浸泡处理48h诱导率最高,诱导率可达13.79%。四倍体与二倍体相比:叶片明显变厚,颜色加深,叶片气孔及其保卫细胞大小增大,气孔密度减小,且保卫细胞内叶绿体数目增加。

引种新资源——印度人参根提取物降血脂作用研究

目的:研究引种新资源印度人参根提取物的降血脂作用。方法:将50只小鼠随机分成阴性对照组(普通饲料)、阳性对照组(脂肪乳剂)和高、中、低剂量组(提取物+脂肪乳剂),分笼喂养30d,测定实验前后血清TC、TG、LDL-C和HDL-C,并处死动物观察肝脏组织病理学变化。结果:印度人参根提取物可明显降低高脂饮食所致高血脂小鼠TC、TG和LDL-C含量(p<0.05),升高HDL-C含量(p<0.05),可降低高脂小鼠肝组织中脂肪细胞数量。结论:引种新资源——印度人参根提取物有辅助降血脂作用。