Antihaemolytic, anti-lipid peroxidative potential by purified protease inhibitors from the fruits of Solanum aculeatissimum Jacq. In human erythrocytes against hydrogen peroxide

Abstract

Protease inhibitor was isolated and purified from the fruits of Solanum aculeatissimum Jacq. (SAPI) via four sequential step procedures i.e., salt precipitation to sepharose affinity chromatography. The purity was confirmed by reverse phase HPLC chromatography. The molecular mass was detected using size elution chromatography (22.2 kDa). It has deep roots in history, being one of the major botanicals used in traditional medicine to treat conditions ranging from diabetes, malaria, to snakebites. This study was conducted to quantitatively evaluate anti-haemolytic activity of purified protease inhibitor from S. aculeatissimum, against hydrogen peroxide induced haemolysis using human erythrocyte in an in vitro assay. Prior to the addition of H2O2 to induce haemolysis, different concentrations (50-500 µg/ml) of SAPI was added to 2 ml of 4% erythrocyte suspension and allowed to incubate for 5 minutes at room temperature. The mixture was centrifuged and the colour density of the supernatant was measured spectro photometrically. Quercetin was used as internal standard. The % haemolysis and IC50 values were calculated. The PI was potent against haemolysis of the erythrocyte in concentration dependent manner with IC50 = 368.54 µg/ml. IC50 value of quercetin was 224 µg/ml. The lower the IC50 the more protection offered against haemolysis by SAPI. These results suggest that the SAPI is ideal anti-haemolytic agent and offered significant biological action compared with standard drug employed.