Protective Effect of Coleus Aromaticus Plant Protein against T-Booh Induced Cell Damage

Abstract

Coleus aromaticus plant is having potential medicinal properties. In this study, we have purified a protein and characterized it by investigating the active protein was purified by G-50 column chromatography and its homogeneity was confirmed with SDS PAGE and RP HPLC. The protein molecular weight was confirmed with MS MALDI. The Phosphomolybdenum assay was used to evaluate the antioxidant capacity, where α- tocopherol was used as standard. The protective effect of purified Coleus aromaticus leaves protein CALP against t-BOOH induced DNA damage was studied using submarine gel electrophoresis, where ascorbic acid is used as the positive control. The trypan blue dye exclusion method was used to find the protective and non toxic nature of protein, where antioxidants like Butylated Hydroxylnisole (BHA) and Ascorbic acid were used as the positive control. The results showed that the molecular weight of Coleus aromaticus plant leaves protein (CALP) is 36 kDa. The purified CALP antioxidant capacity is compared to α- tocopheroland showed maximum protection at a concentration of 25ug.The cell viability study showed that lymphocytes treated with t-BOOH in presence of CALP (25µg) exhibited 69 ± 1% viability, whereas BHA (400μM) had 70 ± 2% viability and ascorbic acid (400µM) had 75 ± 2 % viability when compared to untreated lymphocytes (81 ± 3%) as they are not affected by any external factor like t-BOOH. The purified 36kDa CALP is a nontoxic antioxidant which provides protection against DNA damage induced by t-BOOH.