Suresh Jagtap

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.

Evaluation of comparative free-radical quenching potential of Brahmi (Bacopa monnieri) and Mandookparni (Centella asiatica)

Ayurvedic texts describe rejuvenate measures called Rasayana to impart biological sustenance to bodily tissues. Rasayana acting specifically on brain are called Medhya Rasayana. Brahmi is one of the most commonly practiced herbs for the same. Yet there exist a controversy regarding the exact plant species among Bacopa monnieri L. Penn (BM) and Centella asiatica (L.) Urban (CA) to be used as Brahmi in the formulations. Though the current literature available has suggested a very good nootropic potential of both the drugs, none of the studies have been carried out on comparative potential of these herbs to resolve the controversy. Free-radical scavenging potential for these plants is studied to find out their comparative efficacy. The study revealed a very good in vitro free-radical scavenging properties of aqueous and ethanolic extracts of both the plants as evidenced by FRAP, DPPH, reducing power, and antilipid peroxidation assays. It can be concluded from the studies that both the plants, although taxonomically totally different at family level, showed similar type of in vitro activities. The total phenolic and flavonoid contents also revealed a significant similarity in the two plants. The in vitro study supports the Ayurvedic concept of BM and CA having a similar potential.

Evaluation of the bioavailability of major withanolides of Withania somnifera using an in vitro absorption model system

Withania somnifera (L.) Dunal, shows several pharmacological properties which are attributed mainly to the withanolides present in the root. The efficacy of medicinally active withanolides constituents depends on the absorption and transportation through the intestinal epithelium. We examined these characteristics by employing the Sino-Veda Madin-Darby canine kidney cells culture system, which under in vitro condition shows the absorption characteristics similar to the human intestinal epithelium. Thus, the aim of the present investigation was to assess the bioavailability of individual withanolides. Withanolides were diluted in Hank′s buffered saline at a concentration of 2 μg/ml were tested for permeability studies carried out for 1 h duration. Permeability was measured in terms of efflux pump (Peff ) in cm/s. Peff values of withanolide A (WN A), withanone (WNN), 1,2-deoxywithastramonolide (1,2 DWM), withanolide B (WN B), withanoside IV-V (WS IV-V), and withaferin A were 4.05 × 10−5 , 2.06 × 10−5 , 1.97 × 10−5 , 1.80 × 10−5 , 3.19 × 10−6 , 3.03 × 10−6 and 3.30 × 10−7 respectively. In conclusion, the nonpolar and low molecular weight compounds (WN A, WNN, 1,2 DWM, and WN B) were highly permeable. As against this, the glycosylated and polar WS IV and WS V showed low permeability. Surprisingly and paradoxically, the highly biologically active withaferin A was completely impermeable, suggesting that further studies possibly using human epithelial colorectal adenocarcinoma (Caco-2) cells may be needed to delineate the absorption characteristics of withanolides, especially withaferin A.

Hepatoprotective effect of withanolide-rich fraction in acetaminophen-intoxicated rat: decisive role of TNF-α, IL-1β, COX-II and iNOS

Abstract Context: Overdose of acetaminophen (APAP) is common in humans and is often associated with hepatic damage. Withania somnifera (L.) Dunal (Solanaceae) shows multiple pharmacological activities including antioxidant and anti-inflammatory potential. Objective: To evaluate the possible mechanism of hepatoprotective activity of withanolide-rich fraction (WRF) isolated from a methanolic extract of Withania somnifera roots. Materials and methods: Hepatotoxicity was induced by oral administration of APAP (750 mg/kg, p.o.) for 14 d. The control group received the vehicle. APAP-treated animals were given either silymarin (25 mg/kg) or graded doses of WRF (50, 100 and 200mg/kg) 2 h prior to APAP administration. Animals were killed on 15th day and blood and liver tissue samples were collected for the further analysis. Results: In WRF-treated group, there was significant and dose-dependent (p < 0.01 and p < 0.001) decrease in serum bilirubin, ALP, AST and ALT levels with significant and dose-dependent (p < 0.01 and p < 0.001) increase in hepatic SOD, GSH and total antioxidant capacity. The level of MDA and NO decreased significantly (p < 0.01) by WRF treatment. Up-regulated mRNA expression of TNF-α, IL-1β, COX-II and iNOS was significantly down-regulated (p < 0.001) by WRF. Histological alternations induced by APAP in liver were restored to near normality by WRF pretreatment. Conclusion: WRF may exert its hepatoprotective action by alleviating inflammatory and oxido-nitrosative stress via inhibition of TNF-α, IL-1β, COX-II and iNOS.

Influence of Six Medicinal Herbs on Collagenase-Induced Osteoarthritis in Rats

Medicinal herbs have been effectively used for their anti-inflammatory activity, but their exact role has not yet been documented in scientific literature for the management of Osteoarthritis (OA). Since Sida cordifolia L., Piper longum L., Zingiber officinale Rosc., Ricinus communis L., Vitex negundo L. and Tribulus terrestris L. have been widely used in traditional medicine for their anti-inflammatory activity, to evaluate anti-osteoarthritic activity of these herbs, we used a collagenase type II-induced osteoarthritis (CIOA) rat model. Arthritis was induced in wistar rats by intra-articular injection of collagenase type II. Powders of herbs were given orally for 20 days as a suspension in water (270 mg/kg b. wt.). The effects of the treatment in the rats were monitored by physiological parameters like body weight, knee diameter, paw retraction, paw volume, glycosaminoglycan (GAG) release, radiography and histopathology of knee joint. Selected herbs have significantly prevented body weight loss and knee swelling compared to arthritic control (CIOA). All test groups, including indomethacin (standard drug, 3 mg/kg), significantly reduced paw volume compared to CIOA. GAG release in the serum was significantly lowered in herb treated groups compared to indomethacin. The anterior posterior radiographs of S. cordifolia and P. longum treated groups showed a protective effect against OA. Histopathology revealed protection in the structure of the articular cartilage and in chondrocyte pathology as well as reduced clefting. Treatment with herbs has shown chondroid matrix within normal limits. From the results, we observed that S. cordifolia and P. longum possess potent anti-osteoarthritic activity.

Comparative Hepatoprotective Potential of Tinospora cordifolia, Tinospora sinensis and Neem-guduchi.

Aims :Aim of this study was to evaluate the comparative efficacy of Tinospora cordifolia (Willd.)Miersex Hook. F.,Tinospora sinensis(Lour.) Merrill and T. cordifoliagrowing on Neem (Azadirachta indicaA.Juss.) calledNeem -guduchi. Study Design: Selected species have beenwidely used in the traditional medicine systems in various dosage forms to treat liver disorders. They are of common occurrence and are being used as substitutes to each other. There is no comparative hepatoprotection study yet published, therefore, present study has carried out. Place and Duration of Study:Interactive Research School for Health Affairs, Pune and

Quantification of major bioactive withanolides in Withania somnifera (Ashwagandha) roots by HPTLC for rapid validation of Ayurvedic products

The high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B for the validation of Withania somnifera (Ashwagandha roots) as raw material and Ashwagandha-containing finished Ayurvedic products. HPTLC of W. somnifera methanolic extracts was performed on Si 60F254 (10 × 10 cm) HPTLC plates with dichloromethane-toluene-methanol-acetone-diethyl ether (7.5:7.5: 3:1:1 v/v) as a mobile phase. Upon separation, quantitative evaluation of these withanolides was performed in the absorption reflection mode at 235 nm. The method was validated for precision, reproducibility, and accuracy. On the basis of RF values of 0.58, 0.61, 0.68, and 0.79 for withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B, respectively, were identified. On the basis of linear calibration curves for all withanolides in the range of 0.2−1.2 µg, an average recovery of withaferine A, 1,2 deoxy-withastramonolide, withanolide A, and withanolide B was 98%, 99.5%, 98%, and 99%, respectively. The method is very simple, precise, specific, sensitive, accurate, and economical for rapidly validating the Ayurvedic products containing W. somnifera (Ashwagandha).

Anti-fatigue effect of Amarkand on endurance exercise capacity in rats.

BackgroundAmarkand tubers are routinely used by many Indian tribes as a specialized food for health and longevity but so far there is no scientific evidence for their activities. Taxonomically, Amarkand belong to genera Eulophia and Dioscorea.MethodsIn this communication, comparative antifatigue potential of Amarkand was analyzed using forced swimming model in rats and evaluated using biomarkers of physical fatigue.ResultsMethanol extracts of tubers of D. bulbifera, E. ochreata, E. leghapanensis and bulbils of D. bulbifera exhibited rich polyphenolic content. D. bulbifera bulbils and E. ochreata significantly prolonged the swimming endurance time. Creatine kinase and urea nitrogen were significantly reduced by treatment of D. bulbifera bulbils and E. ochreata as compared to negative control. D. bulbifera bulbils effectively increased creatine (p < 0.001), lactate dehydrogenase (p < 0.01) and hemoglobin (p < 0.001) compared to negative control. D. bulbifera bulbils and E. ochreata treatments significantly increased glycogen (p < 0.05, p < 0.01) and lowered malondialdehyde levels (p < 0.001) in muscles and in liver tissue compared to negative control.ConclusionThese results indicate that a treatment with D. bulbifera bulbils and tubers of E. ochreata facilitates aerobic glucose metabolism and endurance by improving various impairments associated with fatigue.

Amarkand: A comprehensive review on its ethnopharmacology, nutritional aspects, and taxonomy.

In India, the term “Amarkand” is commonly used for around 30 different plant species belonging to genus Eulophia (Orchidaceae). This single local name Amarkand to different taxonomical species creates uncertainty about its ethnomedical and nutritional claims. In the present article, we have reviewed available literature regarding ethnopharmacology, phytochemistry, taxonomy, nutritional, and pharmacological studies of different Amarkand species. The literature was searched using Google Scholar, PubMed, Scopus, and Web of Science databases. Some textbooks and reference books were also used to collect information about traditional and ethnopharmacological records. Amarkand species have been used as a remedy for the treatment of various diseases such as diarrhea, stomach pain, rheumatoid arthritis, cancer, asthma, bronchitis, sexual impotency, tuberculosis, and so on. Nutritionally, Amarkand is considered as an excellent food for children and convalescents. Recent studies confirm antioxidant, anti-inflammatory, anti-diarrheal, and so forth activities to Amarkand species. These species are reported to possess various phytoconstituents such as flavonoids, terpenoids, and phenanthrene derivatives. The present review will help to understand overall ethnopharmacology, nutritional aspects, and taxonomy of Amarkand species.

Effect of macronutrient deficiency on withanolides content in the roots of Withania somnifera and its correlationship with molybdenum content

Abstract Context: The content of withanolides in the roots of Withania somnifera (L.) Dunal (Solanaceae) is important for therapeutic application. Earlier studies have shown that the deficiency of macro- and micronutrients affects the growth of W. somnifera. Therefore, we examined the effect of these deficiencies on the withanolides content of the roots. Objective: To examine the effect of molybdenum accretion in nitrogen-, phosphorus-, calcium- and potassium-deficient soils on the accumulation of withanolides in the roots of W. somnifera. Different withanolides have different therapeutic applications hence major bioactive withanolides assume importance. Materials and methods: Methanol extracts of the roots were subjected to HPTLC and individual withanolides were identified by comparing their Rf values with those of the authentic samples. Molybdenum was quantified by atomic absorption spectroscopy. Free radical scavenging activity was monitored by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay. Results: Molybdenum content in roots of nitrogen-, phosphorus-, calcium-, potassium-deficient, and control plants were 7.02 ± 2.1, 13.1 ± 1.6, 17.1 ± 0.9, 33.5 ± 3.3, and 33.9 ± 1.6 ppm, respectively. Levels of withaferine A increased with the increase in the Mo content in roots from 7.79 ± 2.2 mg/g to 12.57 ± 3.4 mg/g. Antioxidant activity of nitrogen-deficient plants was the lowest (24.7 ± 2.2%) compared to other groups. Discussion and conclusion: It was observed that nitrogen metabolism-dependent molybdenum uptake influences the withanolides accumulation in the roots.