Strengthening antioxidant defense & cardio protection by Piper betle: An in-vitro study

INTRODUCTION: The purpose of this research work was to evaluate Piper betle ethyl acetate extract (PBEA) for its free radical scavenging, antioxidant, anti-apoptotic activities and its role in protecting against oxidative cardiac cell injury. METHODS: The Free radical scavenging activity and antioxidant potential of PBEA were evaluated using various non-cellular methods (1,1-Diphenyl-2-picrylhydrazyl, β-carotene bleaching, superoxide anion, hydroxyl radical, hydrogen peroxide(,) Reducing power, Total phenolics and Total flavonoids). PBEA was standardized with Eugenol by GC-FID analysis. Furthermore, PBEA was also assessed for its cytotoprotective effect against 100μM H(2)O(2) in H9c2 cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Intracellular reactive oxygen species scavenging and anti-apoptoic activity of PBEA was assessed by using 2′, 7′-Dichlorofluorescein diacetate and Annexin- Propidium Iodide, respectively. RESULTS: PBEA exhibited radical scavenging and antioxidant defense response at different magnitudes of potency. Eugenol, a cardiac protective bioactive molecule in PBEA was found to be 43.43 ± 1.46 mg/g of PBEA extract. Further, pre-incubation of H9c2 cells with 10 μg/ml PBEA for 24 h exhibited remarkable cytoprotective effect against H(2)O(2) induced oxidative stress. PBEA at 10 μg/ml dose with 24 h contact with H9c2 cells significantly enhanced the activity of cellular defense system and significantly decreased intracellular ROS (P < 0.001) and apoptosis (P < 0.01) thereby protecting against the cytotoxic effects of H(2)O(2). CONCLUSION: These outcomes indicated that PBEA could shield against oxidative and apoptotic cardiac cell injury in invitro studies. Thus, PBEA might be a desirable antioxidant of natural origin that has future clinical implications in both health care and food industry.