Manganahalli Manjunath Setty

Selective cytotoxicity and pro-apoptotic activity of stem bark of Wrightia tinctoria (Roxb.) R. Br. in cancerous cells

Background: Wrightia tinctoria (Roxb.) R. Br. is a widely available shrub in India used traditionally in various ailments, including cancer. However, the anticancer activity of the bioactive fractions has not been validated scientifically. Objective: To investigate the anticancer potential of stem bark of W. tinctoria and establish its phytochemical basis. Materials and Methods: The ethanol extract and subsequent fractions, petroleum ether, ethyl acetate, n-butanol, and aqueous were prepared by standard methods. In vitro cytotoxicity was determined in MCF-7 (breast) and HeLa (cervical) adenocarcinoma cells, and V79 (nontumor fibroblast) cells and apoptogenic activity in MCF-7 cells by acridine orange (AO)/ethidium bromide (EB) staining. Additionally, the antioxidant potential was evaluated using suitable methods. High-performance thin layer chromatography (HPTLC) analysis was performed for identification of active phytoconstituents. Results: Petroleum ether and ethyl acetate fractions were most potent with IC50 values of 37.78 and 29.69 μg/ml in HeLa and 31.56 and 32.63 μg/ml in MCF-7 cells respectively in the sulforhodamine B assay. Comparable results were obtained in HeLa cells in 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide assay and interestingly, the fractions were found to be safe to noncancerous fibroblast cells. Both fractions induced significant (P < 0.05) apoptotic morphological changes observed by AO/EB staining. Moreover, extract/fractions exhibited excellent inhibition of lipid peroxidation with the ethyl acetate fraction being most active (IC50:23.40 μg/ml). HPTLC confirmed the presence of two anti-cancer triterpenoids, lupeol, and β-sitosterol in active fractions. Conclusion: Extract/fractions of W. tinctoria exhibit selective cytotoxicity against cancerous cells that is mediated by apoptosis. Fractions are less toxic to noncancerous cells; hence, they can be developed as safer chemopreventive agents. SUMMARY Petroleum ether and ethyl acetate fractions were most active and exhibited dose-dependent cytotoxicity in HeLa and MCF-7 cells. Fractions were relatively less toxic to non-tumor fibroblast cells demonstrating its selectivity to cancer cells. Fractions exhibited pro-apoptotic activity in MCF-7 cells in AO/EB staining. Lupeol and β-sitosterol were identified as anticancer constituents by HPTLC.

Evaluation of antiurolithiatic and antioxidant potential of Lepidagathis prostrata: A Pashanbhed plant

Abstract Context: Oxidative stress acts as an essential mediator in the pathophysiology of urolithiasis. Lepidagathis prostrata Dalz. (Acanthaceae) is a Pashanbhed plant that is recommended for the management of urolithiasis; however, no scientific validation has been reported. Objectives: To evaluate the antiurolithiatic and antioxidant potential of L. prostrata. Materials and methods: Methanol extract (LPM) and fractions; petroleum ether (LPPE), ethyl acetate (LPEA), n-butanol (LPBU) and aqueous (LPAQ) were prepared. In vitro antiurolithiatic activity was evaluated by the capacity to inhibit calcium oxalate (CaOx) nucleation and aggregation at different concentrations of extract/fractions (0.04–3 mg/mL) for 30 min. Total phenol and flavonoid content and antioxidant potential were determined. A validated HPTLC method was performed to quantify lupeol and β-sitosterol. Results: LPEA exhibited the highest dose-dependent inhibition of CaOx nucleation (IC50: 336.23 ± 30.79 µg/mL) and aggregation (IC50: 149.63 ± 10.31 µg/mL), which was significantly (p < 0.05) better than standard Cystone®. The polar LPBU fraction was enriched with phenols (47.34 ± 0.19 mg GAE/g) and flavonoids (20.38 ± 0.05 mg QE/g), which correlates with its highest antioxidant potential in DPPH, ABTS, nitric oxide scavenging and iron chelating activities (IC50: 1.18–87.34 µg/mL). To our knowledge, this is the first study reporting the presence of lupeol and β-sitosterol in L. prostrata. Conclusion: The antiurolithiatic activity of L. prostrata is probably mediated through the inhibition of CaOx crystallization. In addition to its free radical scavenging and antioxidant activities, it would act as an excellent agent for the prevention of urolithiasis.

Anticancer activity of stem bark extract and fractions of wrightia tinctoria in transplantable tumors in mice

Wrightia tinctoria Roxb. (Apocynaceae), widely used in traditional medicine for inflammatory conditions and cancer, has not been studied extensively for its anticancer potential. The present study evaluated the anticancer activity of ethanol extract (EE) and petroleum ether (PE) and ethyl acetate (EA) fractions of stem bark of W. tinctoria using in vivo models. The anticancer activity was evaluated in Dalton’s lymphoma ascites (DLA) solid tumor and Ehrlich’s ascites carcinoma (EAC) liquid tumor models after treatment for 18 and 13 days respectively at 50 and 100 mg/kg i.p. EA at 100 mg/kg was most effective in DLA model with 71.1% inhibition in tumor volume. In EAC model, EA significantly inhibited body weight gain due to liquid tumor (p < 0.05). The mean survival time (MST) increased (p < 0.001) at both doses along with significant improvement in hematological, and antioxidant parameters. W. tinctoria possesses significant anticancer activity and further investigations are in progress to identify the phytochemical and mechanistic basis of action.

Evaluation of In Vitro Antiurolithiatic Potential of Bridelia retusa Spreng. Bark

Urolithisis is a very complex disease which is characterized by the formation of renal calculi in urinary track/kidney. Several physic chemical events like super saturation, nucleation, growth, aggregation and retention of dietary minerals of urine in the kidney leads to the formation of kidney stone; for this a essential need to develop a formulation which is made up of medicinal pant belong to the class of Pasanabheda plant according to ancient literature available in Ayurveda, which facilitate the repair mechanism in a natural way towards the organ. Bridelia retusa, is a well know plant of Western Ghats. Present study deals with the inhibitory effect of different fraction of methanol extract of Bridelia retusa. Nucleation of calcium oxalate crystals was estimated by the spectrophotometric assay and whereas crystal dissolution was determine by titrimetry/egg shell permeation method, hence the extract fractions exhibited inhibitory action in both of nucleation and aggregation assays to significant level.