In-vitro cytoprotective activity of Eichhornia crassipes ϐlowers against hydrogen peroxide induced oxidative stress in BRL 3A rat liver cells

Abstract

The aim of this study was to see whether an ethanol extract of Eichhornia crassipes flowers and its fractions could protect BRL 3A liver cells from hydrogen peroxide-induced oxidative stress. Eichhornia crassipes powdered flowers were subjected to a hot continuous extraction in a soxhlet extractor using ethanol as the solvent material. The solvent extracts were first tested for in-vitro free radical scavenging and anti-oxidant activity using qualitative and quantitative methods. Benzene, chloroform, and n-butanol were used to fractionate the ethanol extract. In BRL 3A cell lines, the crude ethanol extract and its fractions were tested for their possible cytoprotective effect against hydrogen peroxide (H2O2) induced oxidative stress. Cell viability, lipid peroxidation by measuring the formation of malondialdehyde, lactate dehydrogenase leakage into culture medium, catalase activity, and the content of reduced glutathione (GSH) in the cells were all tested in biochemical assays to determine the cytoprotective activity. BRL 3A cells were exposed to 2mM H2O2, which decreased cell viability, increased malondialdehyde (MDA) levels, increased lactate dehydrogenase (LDH) leakage, and reduced antioxidant activities. Pretreatment of cultured cells for 30 minutes with crude ethanol extract of Eichhornia crassipes flowers and various solvent fractions at concentrations of 0.01, 0.1, 1, 10, 100 g/ml attenuated oxidative injury in a dose-dependent manner until H2O2 exposure. The crude ethanol extract of Eichhornia crassipes flowers was found to have a strong cytoprotective impact, raising cell viability while decreasing lipid peroxidation and LDH leakage. In the cells pre-treated with ethanol extract of Eichhornia crassipes flowers, there was a further increase in catalase and a decrease in glutathione activity. These results indicate that an ethanol extract of Eichhornia crassipes flowers has potent cytoprotective properties against reactive oxygen species-induced oxidative injury.