Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Inhibitory effects of total saponins from Ilex pubescens Hook against hydrogen peroxide-induced cardiomyocyte apoptosis

Ming-zhong Ying, Jian-jun Wu, Jian-min Yin, Xue-qin Shen, Fan Li

The International Medical Center of the Chinese PLA General Hospital, Beijing 100853, China;

For correspondence:- Fan Li Email: lifanpla@126.com

Accepted: 10 October 2017 Published: 30 November 2017

Citation: Ying M, Wu J, Yin J, Shen X, Li F. Inhibitory effects of total saponins from Ilex pubescens Hook against hydrogen peroxide-induced cardiomyocyte apoptosis. Trop J Pharm Res 2017; 16(11):2681-2688 doi: 10.4314/tjpr.v16i11.16

© 2017 The authors.
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Abstract

Purpose: To study the protective effects of total saponins from Ilex pubescens Hook (IPTS) against cardiomyocyte apoptosis.

Methods: Response surface methodology (RSM) based on Box-Benhnken Design (BBD) was carried out to optimize the extraction of IPTS. Thereafter, H9c2 cell model prepared by hydrogen peroxide (H₂O₂) treatment was used to investigate the effects of IPTS on cardiomyocyte apoptosis. Cell viability was determined using MTT assay, while the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), creatine kinase (CK) and catalase (CAT) were measured as indices of oxidative stress. expressions of proteins related to apoptosis (caspase-3, Bax and Bcl-2) were measured using Western blot assay.

Results: Optimal IPTS extraction was achieved with extraction temperature of 86.6 °C, extraction time of 2.23 h and water: raw material ratio of 10.8 mL/g. IPTS extract, at doses of 200, 400, 600 and 800 μg/mL, significantly increased the viability of H₂O₂-treated H9c2 cells (p < 0.05), but significantly decreased LDH and CK activities (p < 0.01). It also led to significant increases in SOD and CAT activities, and significant decreases in the levels of MDA in these cells (p < 0.01). There were significant down-regulation of the protein expressions of caspase-3 and Bax (p < 0.01) in IPTS-treated H9c2 cells, as well as significant up-regulation of Bcl-2 protein expression (p < 0.01).

Conclusion: These results suggest that IPTS can protect cardiomyocytes against apoptosis via the inhibition of oxidative stress and mitochondria-induced intrinsic apoptosis

Keywords: Ilex pubescens, Total saponins, Cardiomyocytes, Apoptosis, H9c2 cells