In-vitro Cytoprotective Activity of Eichhornia crassipes Flowers Against Hydrogen Peroxideinduced Oxidative Stress in BRL 3A Rat Liver Cells

Abstract

The aim of this study was to investigate the cytoprotective effect of ethanol extract of Eichhornia crassipes flowersand its fractions against hydrogen peroxide induced oxidative stress in BRL 3A liver cells. Powdered flowers of Eichhornia crassipeswere subjected to hot continuous extraction in soxhlet extractor using ethanol as solvent material. Initially, the solvent extracts were subjected to qualitative, quantitative analysis and assessed for in-vitro free radical scavenging activity and anti-oxidant activity. The ethanol extract was fractionated using benzene, chloroform and n-butanol. The crude ethanol extract and its fractions were evaluated for its potential cytoprotective effect against hydrogen peroxide (H2O2) induced oxidative stress in BRL 3A cell lines.Biochemical assays were carried out to determine the cytoprotective activity, including cell viability, lipid peroxidation by determining the formation of malondialdehyde, lactate dehydrogenase leakage into culture medium, the catalase activity and the content of reduced glutathione (GSH) in the cells. Exposure of BRL 3A to 2mM H2O2 reduced the cell viability, increased the malondialdehyde (MDA) level, increased the leakage of lactate dehydrogenase (LDH) and caused reduction in antioxidant activities. Pretreatment of cultured cells with crude ethanol extract of Eichhornia crassipes flowers and different solvent fractions at concentrations 0.01, 0.1, 1, 10, 100 μg/ml for 30 minutes before H2O2 exposure attenuated the oxidative injury in dose-dependent manner. It was observed that crude ethanol extract of Eichhornia crassipes flowers exhibited a strong cytoprotective by increasing cell viability, decreasing lipid peroxidation and LDH leakage. Further increase in catalase and reduced glutathione activity was noted in the cells pre-treated with ethanol extract of Eichhornia crassipes flowers. These findings suggest that ethanol extract of Eichhornia crassipes flowers has a strong cytoprotective activity against oxidative injury caused by reactive oxygen species.