Cardioprotective effect of Butanol Fraction of Rivea ornata against H2O2 Induced Oxidative Stress in H9c2 Cells: Role of its Phenolic Content and Antioxidant Effect

Abstract

Rivea ornata used traditionally for various disease of heart. However, its protective effect against oxidative stress induced injury is unclear and unproved. So, we attempted to investigate the protective role of butanol fraction of Rivea ornata on H9c2 cardiomyocytes against oxidative stress induced by H2O2. Ethanol extract and its fractions such as n-hexane, ethyl acetate and butanol fractions were assessed for its phytochemical composition, total phenolic and flavonoid content and assessed in vitro antioxidant activity by using established assays such as DPPH, H2O2 neutralization activity, ferric reduction assays and total antioxidant capacity. Among all the extracts/fractions butanol fraction found with high phenolic (82.4 ± 4.61 mg of GAE/g) and flavonoid (105 ± 4.09 mg of QE/g) content as well as possess strong antioxidant abilities evident by comparative lowest IC50 values for DPPH (41.80 µg/mL), H2O2 neutralization assay (35.39 µg/m), highest ferric reduction at 120µg/mL and high antioxidant capacity equivalent to quercetin (72.5±2.98 µg/mL). By considering these results, only butanol fraction assessed for its protective effect H2O2 induced oxidative stress on H9c2 cardiac cell lines was assessed by MTT, LDH assay and ROS expression studies. Butanol fraction at 25µg/mL maintain H9c2 cell lines viability (in MTT assay), cell integrity (decreased LDH release) and decreased ROS expression (decrease in percentage mean fluorescence). These findings are proved antioxidant rich polyphenolic butanol fraction protective effect against oxidative stress injury observed with H2O2 by maintain cell viability, cell integrity and ROS expression.