A Chinese 4-herb formula, Yiqi-Huoxue granule, alleviates H2O2-induced apoptosis by upregulating uncoupling protein 2 in H9c2 cells

Abstract

Background: Although the protective effects of Yiqi-Huoxue granule (YQHX), a Chinese 4-herb formula, on patients with ischemic heart diseases are related to the attenuation of oxidative stress injury, the mechanism(s) underlying these actions remains poorly understood.

Purpose: Our aim was to investigate the potential protective effects of YQHX treatment against oxidative stress induced by hydrogen peroxide (H₂O₂) in rat H9c2 cells.

Methods: H9c2 cells were treated with YQHX for 16 h before exposed to 200 μM H₂O₂ for 6 h. The apoptosis induced by H₂O₂ was measured using hoechst 33,342 staining and Annexin-V FITC/PI assay. The expression of uncoupling protein 2 (UCP2), Bcl-2, Bax, and caspase-3 were observed using western blot. The effects of UCP2 knockdown on cell apoptosis and intracellular ROS production were also investigated.

Results: H₂O₂ exposure led to significant activation of oxidative stress followed by increased apoptosis and ROS production, as well as decreased UCP2 expression in H9c2 cells. YQHX treatment at the concentration of 0.75 and 1.5 mg/ml remarkably reduced the expression of Bax and caspase-3, whereas increased the protein expression of Bcl-2 and UCP2. These changes were attenuated by transgenic knockdown of UCP2 with Lenti-shUCP2 vector.

Conclusions: Taken together, our study demonstrated that YQHX attenuates H₂O₂-induced apoptosis by upregulating UCP2 expression in H9c2 Cells, suggesting that YQHX is a promising therapeutic approach for the treatment of I/R injury-mediated apoptosis.

Keywords: Cardiomyocyte apoptosis; Oxidative stress; Uncoupling protein 2 (UCP2); Yiqi-Huoxue granule (YQHX).