Antioxidant and Nephroprotective Activities of the Extract and Fractions of Homonoia riparia Lour.

BACKGROUND: Homonoia riparia is a plant, which is widely used in the indigenous system of medicine for the treatment of urolithiasis, renal disorders and inflammatory conditions. This is the first report on the antioxidant and nephroprotective activities of whole plant of H. riparia. OBJECTIVE: The present study aims at investigating the in vitro antioxidant and nephroprotective activity of the methanol extract and its different fractions of H. riparia. METHODS: Petroleum ether (HRPE), Ethyl acetate (HREA), Butanol (HRBU), aqueous fractions (HRAQ) were prepared from the crude methanol extract of H. riparia (HRM) using liquid partitioning. Total phenolic content, flavonoid content and antioxidant activity assay were performed according to suitable methods. Nephroprotective activities were evaluated by MTT assay using Human Embryonic Kidney cells against cisplatin induced toxicity. Quantification of gallic acid was performed using validated HPTLC method. RESULTS: The studies showed that extract and fractions possess significant nephroprotective activity against cisplatin induced renal toxicity. All the extracts/fractions of whole plant of Homonoia riparia was found to be significantly reducing cisplatin induced toxicity (< 0.05). The highest activity was observed with HRBU and HRAQ with a percentage viability of 293.09 ± 4.3 and 345.07 ± 3.2 at a concentration of 200 µg/ml. Gallic acid was detected in the HRM/fractions using HPTLC. SUMMARY: Cisplatin (8 µg/ml) exhibited 50 % inhibition in cell viability in HEK 293 cellsButanol and aqueous fractions of Homonoia riparia showed significant nephroprotective activity against cisplatin induced cell damage in HEK cells.Gallic acid was detected and quantified in the extract and fractions of whole plant of Homonoia ripariaAbbreviations used: HPTLC: High Performance Thin Layer Chromatography, DPPH: 1,1-diphenyl-2-picrylhydrazyl, ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, MTT: 3-(4,5-dimethylthiazolyl-2-yl)-2,5- diphenyl tetrazolium bromide, GAE: Gallic acid equivalents, QE: Quercetin equivalents, HEK: Human Embryonic Kidney, HRM: Methanol extract of H. riparia, HRPE: Petroleum ether fraction of H. riparia, HREA: ethyl acetate fraction of H. riparia, HRBU: Butanol fraction of H. riparia, HRAQ: Aqueous fraction of H. riparia, DMEM: Dulbecco's minimum essential medium, FBS: Foetal bovine serum, DMSO: Dimethyl sulfoxide, ANOVA: One way analysis of variance.

Evaluation of Ceiba pentandra (L.) Gaertner bark extracts for in vitro cytotoxicity on cancer cells and in vivo antitumor activity in solid and liquid tumor models.

The stem bark of Ceiba pentandra (L.) Gaertner is claimed to be useful in the treatment of tumors in the southern part of India. This plant possesses a number of sesquiterpenoids and isoflavones which are known for their anticancer properties. The present study was designed to scientifically evaluate the cytotoxic potential of bark extracts in in vitro on Ehrlich ascites carcinoma (EAC), MCF-7 and B16F10 cells and in vivo in EAC (Liquid tumor) model and Dalton's lymphoma ascites (DLA or solid tumor) model. The bark was powdered and extracted successively with solvents viz., petroleum ether (PE), benzene, chloroform, acetone (AC), and ethyl alcohol in the sequential order of polarity. Cytotoxicity of dried extracts was screened on EAC cells by trypan blue assay. Three potent extracts namely petroleum ether, acetone, and ethanol were screened for their cytotoxicity on MCF-7 and B16F10 cells by MTT assay and nucleomorphological alteration by propidium iodide staining. Safe doses of these extracts were evaluated by acute toxicity study in mice. Extracts were found to be safe up to 300 mg/kg in acute toxicity study. Dosage of 1/10th and 1/20th of safe dose i.e., 15 and 30 mg/kg were selected for in vivo study. In the EAC model, both doses of the extracts showed a significant (P < 0.05) improvement in mean survival time and a maximum decline in tumor induced increase in body weight (an indirect measure of tumor weight) by the PE and AC treatment at 15 mg/kg compared to control. In the DLA-model, all extracts at both tested dose levels showed >50 % reduction in tumor weight and a significant reduction (P < 0.05) in tumor volume on the 30th day compared to control. It can be concluded that these extracts possess cytotoxic and antitumor activity.

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

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 ß-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 ß-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.

Evaluation of antioxidant and anticancer activity of extract and fractions of Nardostachys jatamansi DC in breast carcinoma.

BACKGROUND: Nardostachys jatamansi DC is a Himalayan medicinal herb that has been described in various traditional systems of medicine for its use in cancer. In view of its traditional claims, and chemical constituents, antioxidant and anticancer activities were evaluated in breast carcinoma. METHODS: Petroleum ether (NJPE), methanol extract (NJM) and subsequent diethyl ether (NJDE), ethyl acetate (NJEA) and aqueous (NJAQ) fractions of roots and rhizomes of N. jatamansi were prepared. Total phenolic, flavonoid content, and antioxidant activities were determined using suitable methods. Antiproliferative activity was assessed in estrogen receptor (ER)-positive (MCF-7) and ER-negative breast carcinoma (MDA-MB-231) cells by MTT and SRB assay. Cell cycle analysis, Hoechst staining, and clonogenic assay were employed to determine the mode of antiproliferative and pro-apoptotic activity in MDA-MB-231 cells. RESULTS: NJM/fractions exhibited prominent antioxidant activity with significant correlation between phenolic content and ABTS (IC50) scavenging (R = -0.9680, P < 0.05), and total antioxidant capacity (R = 0.8396, P > 0.05). In MTT assay, NJM exhibited the highest antiproliferative activity (IC50: 58.01 ± 6.13 and 23.83 ± 0.69 µg/mL in MCF-7 and MDA-MB-231 respectively). Among the fractions, NJPE and NJDE were found to be most potent in MCF-7 (IC50: 60.59 ± 4.78 µg/mL) and MDA-MB-231 (IC50: 25.04 ± 0.90 µg/mL) cells respectively. Statistical analyses revealed NJM and NJDE exhibited significantly higher (P < 0.05) cytotoxicity in MDA-MB-231 cells. Cell cycle analysis demonstrated that NJM, NJPE and NJEA caused G2/M arrest while NJDE caused G0/G1 phase arrest in MDA-MB-231 cells. Further, NJM/fractions induced significant (P < 0.001) cell death by apoptosis characterized by apoptotic morphological changes in Hoechst staining and inhibited long-term proliferation (P < 0.001) of MDA-MB-231 cells in clonogenic assay. Lupeol and ß-sitosterol were identified as anticancer principles in NJM/fractions by HPTLC. CONCLUSION: Our results suggest that NJM/fractions possess significant antiproliferative potential which is mediated through cell cycle perturbation and pro-apoptotic effects in MDA-MB-231 cells. Moreover, this study highlights the antioxidant potential of NJM/fractions which can be attributed to the presence of phenols. NJDE emerged as the most potent fraction and further mechanistic and phytochemical investigations are under way to identify the active principles.

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 ß-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 ß-sitosterol were identified as anticancer constituents by HPTLC.