Effects of Escin on Oxidative Stress and Apoptosis of H9c2 Cells Induced by H ₂O ₂

Abstract

Objective

Myocardial infarction (MI) is a serious heart health problem in the world with a high mortality rate. Our study is mainly aimed at validating the antioxidative stress and antiapoptotic effects of escin in a H ₂O ₂-induced cardiomyocyte injury model.

Methods

H9c2 cells were divided into control group, H ₂O ₂ treatment group, and H ₂O ₂+escin group. We studied the effect of escin on H9c2 cells and its mechanism by flow cytometry, real-time PCR, CCK-8 assay and Western blot. Cell morphology was observed by cell staining and optical microscopy.

Results

We found that the level of reactive oxygen species (ROS) in the H ₂O ₂ treatment group was significantly elevated, while the high level of ROS was significantly reversed after treatment with escin. The protein levels of SOD1, SOD2, Bcl-2, and I κB- α in the H ₂O ₂ treatment group were significantly decreased compared with the H ₂O ₂+escin group, and the Bax, TNF- α, IL-1 β, p65, and I κK α protein expressions were greatly higher than those in the H ₂O ₂+escin group. And the results of PCR were also consistent with those. TUNEL-positive cells also decreased significantly when treated with escin. Flow cytometry showed that the percentage of apoptotic cells decreased greatly after treatment of escin. Through IL-1 β immunofluorescence, the fluorescence intensity of the H ₂O ₂ treatment group was greatly higher compared with that of the control group, but escin reversed this effect.

Conclusions

These results indicated that escin inhibits H ₂O ₂-induced H9c2 cell apoptosis, oxidative stress, and inflammatory responses via the NF- κB signaling pathway.