Identification and quantitative estimation of Emodin in Rhei Emodi Wall.Rhizome and the relevant polyherbal ayurvedic formulation

The present study explored quality control of plant drug and polyherbal formulation using HPTLC and HPLC fingerprinting.The study combined qualitative analysis of Emodin and Rhein biomarker compound in Rhei Emodi Wall.rhizome extract and polyherbal formulation extract by HPTLC method with quantitative estimation of Emodin by HPLC method,aiming to help in accurate identification of Emodin and Rhein in plant materials,polyherbal formulations.This study also checked adulteration,with the purpose of improving bio-efficacy of ayurvedic formulation.All the samples showed characteristic peaks of Emodin at same retention time as that of standard Emodin.According to the HPTLC fingerprinting results,the Rf values of Rhein reference standard at 0.42 and Emodin at 0.62 were observed in test solution of Rhei emodi Wall.rhizome extract and polyherbal formulation extract and both were found comparable.The quantitative evaluation of Emodin present in the polyherbal formulation and Rhei emodi Wall.rhizome extracts were 0.0177% and 0.3106% respectively.

Evaluation of the effect of shodhana(detoxification)process using chromatographic profiling(HPTLC,HPLC,LC-MS,and GC-MS)and estimation of the content of toxic colchicine in Gloriosa superba Linn.tubers

Shodhana(purification/detoxification)is an important aspect of Ayurveda.It has been described in several important pieces of literature.In this study,we performed shodhana process on Langali(Gloriosa superba Linn.)tubers of family Liliaceae,a semi-woody herbaceous climber with significant therapeutic properties.We compared the unprocessed(before purification)and processed(after purification)samples based on the phytochemical,physicochemical,and chromatographic study,including HPTLC,HPLC,GC-MS and LC-MS.Significant changes were observed in profiling after the shodhana process.This plant contains colchicine,known for its toxic effects,particularly cardiotoxicity.Therefore,we quantified colchicine in the unprocessed and processed samples of Gloriosa superba Linn.tubers using HPLC method.Results showed that the percentage of colchicine was reduced by 13.67%in chloroform extract and 20.97%in ethanol extracts after the shodhana process.

Quantitative analysis of catechins in Saraca asoca and correlation with antimicrobial activity

Herbal medicines are highly complex and have unknown mechanisms in diseases treatment. Saraca asoca （Roxb.）, De. Wild has been recommended to treat gynecological disorders and used in several commercial polyherbal formulations. In present study, efforts have been made to explore antimicrobial activity and its co-relation with the distributions of catechins in the organs of S. asoca using targeted MS/MS. Eight extracts （cold and hot water） from four different organs of S. asoca and two drugs were prepared and antimicrobial activity was assessed by microbroth dilution assay. Quantitative and qualitative analysis of catechins in crude extracts was done by using targeted and auto-MS/MS and correlated with antimicrobial activity. （＋）-Catechin and （＋）- epicatechin and their biosynthesis related compound were found to be up-regulated in regenerated bark and leaves extracts. （-）-Epigallocatechin was found to be significantly higher in bark water extract as compared to others but showed low antimicrobial activity. Result showed down- regulation of （-）-epigallocatechin and up-regulation of （＋）-catechin and （＋）-epicatechin in the regenerated bark and leaves of S. asoca. It might be the contributing factor in the antimicrobial activity of regenerated bark and leaves of the plant. The concentration of （＋）-epicatechin in processed drugs （Ashokarishta） from Baidyanath was found to be seven times higher than that of Dabur Pvt. Ltd., but no antimicrobial activity was observed, indicating the variations among the plant based drugs. This will be helpful in rational use of S. asoca parts. Furthermore, the analyticalmethod developed is sensitive, repeatable and reliable; therefore, it is suitable for quality control of herbal drugs.

Anti-atherosclerotic activity of root bark of Premna integrifolia Linn.in high fat diet induced atherosclerosis model rats

Premna integrifolia Linn. is a medicinal plant used in ＂Dhasamula＂ drug preparation of Ayurvedic systems of medicine in the treatment of various ailments like bronchitis, dyspepsia, liver disorders, piles, constipation,hyperlipidemia and fever. The anti-atherosclerotic activity of hydroalcoholic extract（HAE） of root bark of P.integrifolia was evaluated in high fat diet induced atherosclerosis rats. Sixty Wistar rats were divided into six groups： the first group served as control, the second group was fed with high fat diet and the other three groups were fed with high fat diet along with various concentrations of HAE and the last group was treated with atorvastatin for 30 days. Lipid and lipoprotein profile, atherogenic index, and cardiac markers and histopathological evaluation of aorta were determined in high fat diet induced atherosclerosis rats. HAE of P.integrifolia produced a significant and dose-dependent anti-atherosclerotic activity in terms of reduction in lipids and lipoprotein profile, atherogenic index, HMG-Co A reductase activity, marker enzymes such as lactate dehydrogenase（LDH）, creatine phosphokinase（CPK）, aspartate transaminase（AST）, alanine transaminase（ALT） and alkaline phosphatase（ALP）, alteration in collagen and calcium contents, mild mineralization and focal rupture of intima and media of aorta was noticed in treated groups as compared to the control. The results suggested that anti-atherosclerotic activity of HAE of P. integrifolia Linn. was due to its modulatory activity on metabolic pathway of lipid. The results contribute to the validation of the traditional use of Agnimantha in high fat diet induced atherosclerosis rats.

Anti-cancer effects of sitagliptin,vildagliptin,and exendin-4 on triple-negative breast cancer cells via mitochondrial modulation

Triple-negative breast cancer(TNBC)cell line MDA-MB-231 is known for Warburg metabolism and defects in mitochondria.On the other hand,dipeptidyl peptidase-IV(DPP-IV)inhibitors such as sitagliptin and vildagliptin and GLP-1 agonist exendin-4 are known to improve mitochondrial functions as well as biogenesis,but no study has evaluated the influence of these drugs on mitochondrial biogenesis on metastatic breast cancer cell line.We have recently reported anticancer effects of 5-aminoimidazole-4-carboxamide riboside on MDA-MB-231 cells via activation of AMP-dependent kinase(AMPK),which activates the downstream transcription factors PGC-1α,PGC-1β,or FOXO1 for mitochondrial biogenesis;above-mentioned incretin-based therapies are also known to activate AMPK.This study evaluated the effects of sitagliptin,vildagliptin,and exendin-4 on MDA-MB-231 cells and the underlying changes in mitochondrial biogenesis,were examined.Treatment with sitagliptin(100μM),vildagliptin(100μM),and exendin-4(10 nM)for 72 h to MDA-MB-231 cells led to a decrease in viability indicated by MTT assay,cell migration by scratch,and transwell migration assays,accompanied with marginal reduction in cell numbers along with the apoptotic appearance,the rate of apoptosis,and decreased lactate content in conditioned medium.These changes in the cancer phenotype were accompanied by an increase in the mitochondrial DNA to nuclear DNA ratio,increased MitoTracker green and red staining,and increased expression of transcription factors PGC-1α,NRF-1,NRF-2,TFAM,and HO-1.Pre-treatment of cells with these incretin-based drugs followed by 48 h treatment with 1μM doxorubicin increased doxorubicin sensitivity as observed by a decrease in viability by MTT assay.Thus,sitagliptin,vildagliptin,and exendin-4 exert their beneficial effects on TNBC cells via an increase in mitochondrial biogenesis that helps to switch Warburg metabolism into anti-Warburg effect.Therapeutic response was in the order of:sitagliptin>vildagliptin>exendin-4.

Cytotoxic and antiproliferative activity of kanchnar guggulu,an Ayurvedic formulation

Objective： Kanchnar guggulu is a compound Ayurvedic formulation used in clinical practice for the treat- ment of benign and malignant tumors. The present study investigates its cytotoxic and antiproliferative activities.Methods： The hydro-alcoholic （50%） extract of kanchnar guggulu was prepared. Its antimitotic activity was assessed in an Allium cepa assay, while its antiproliferative effects were studied in a yeast prolifer- ation model. Methotrexate was used as a standard anticancer agent.Results： In the Allium assay, all concentrations of the extract （1, 2 and 3 mg/mL） and methotrexate （0.02 mg/mL） significantly inhibited the division ofA. cepa root cells, decreasing root growth and mitotic index compared to control; this effect was concentration-dependent for kanchnar guggulu extract. In the antiproliferative studies, treatment with the hydro-alcoholic extract of kanchnar guggulu （1, 5 and 10 mg/mL） and methotrexate （0.025, 0.05 and 0.1 mg/mL） resulted in marked reduction of dividing Saccharornyces cerevisiae cells and inhibition of cell viability compared to control. The cytotoxicity of the hydro-alcoholic extract of kanchnar guggulu, shown by its antimitotic and antiproliferative effects, may be due to the presence of flavonoids and phenolics.Conclusion： Kanchnar guggulu exhibited a cytotoxic effect by inhibiting cell division （antimitotic） and reducing cell proliferation. These results substantiate its potential for the treatment of cancer and support its traditional use in the treatment of cancer.

Hepatoprotective and antioxidant activity of Bombax ceiba flowers against carbon tetrachloride-induced hepatotoxicity in rats

Aim:The flowers of Bombax ceiba are traditionally used as home remedy in the treatment of jaundice and spleen enlargement.The present work investigated the effect of aqueous extract of flowers of Bombax ceiba(BCAE)on experimentally induced hepatotoxicity in rats to substantiate its traditional use as hepatoprotective agent.Methods:Hepatotoxicity was induced in rats by carbon tetrachloride(CCl_(4))treatment;at the same time vehicle or BCAE(250 or 500 mg/kg)or silymarin(25 mg/kg)were administered daily orally for seven days.Hepatotoxicity was assessed by estimating the activities of marker enzymes and by histological studies.The antioxidant effect of BCAE was assessed by measuring amount of antioxidant phytochemicals(total phenolics and flavonoids),and DPPH free radical scavenging assay of the extract.Results:BCAE treatment significantly prevented the CCl_(4)-induced elevations in levels of glutamate oxaloacatate transaminase,glutamic pyruvic transaminase,alkaline phosphatase,bilirubin,and triglycerides,and decreased the total protein levels.Treatment with BCAE attenuated the CCl_(4)-induced cytotoxic damage to liver.BCAE exhibited presence of antioxidant phytochemicals and showed scavanging action on DPPH radicals.The hepatoprotective effect of BCAE was comparable to that of the standard antioxidant hepatoprotective agent,silymarin.These findings indicated that BCAE showed hepatoprotective effect against CCl_(4)-induced hepatotoxicity and exhibited in vitro antioxidant effects.Conclusion:Bombax ceiba flowers exhibited hepatoprotective effect which may be attributed to antioxidant potential.This study also validated their traditional medicinal use in liver disorders.