Laboratory of Cell Imaging, Henan University of Chinese Medicine, 6 Dongfeng Rd, Zhengzhou, Henan 450002, China; Institute of Cardiovascular Disease, Henan University of Chinese Medicine, Zhengzhou, 450002, China.
Laboratory of Cell Imaging, Henan University of Chinese Medicine, 6 Dongfeng Rd, Zhengzhou, Henan 450002, China.
Phytomedicine. 2019 Feb;53:171-181. doi: 10.1016/j.phymed.2018.09.031. Epub 2018 Sep 5.
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.
Our aim was to investigate the potential protective effects of YQHX treatment against oxidative stress induced by hydrogen peroxide (HO) in rat H9c2 cells.
H9c2 cells were treated with YQHX for 16 h before exposed to 200 μM HO for 6 h. The apoptosis induced by HO 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.
HO 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.
Taken together, our study demonstrated that YQHX attenuates HO-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.
虽然中药四君子方益气活血颗粒（YQHX）对缺血性心脏病患者的保护作用与减轻氧化应激损伤有关，但这些作用的机制仍不清楚。
本研究旨在探讨 YQHX 对过氧化氢（HO）诱导的大鼠 H9c2 细胞氧化应激的潜在保护作用。
用 YQHX 处理 H9c2 细胞 16 h 后，用 200 μM HO 处理 6 h。用 hoechst 33，342 染色和 Annexin-V FITC/PI 法检测 HO 诱导的细胞凋亡。用 Western blot 观察解偶联蛋白 2（UCP2）、Bcl-2、Bax 和 caspase-3 的表达。还研究了 UCP2 敲低对细胞凋亡和细胞内 ROS 产生的影响。
HO 暴露导致氧化应激显著激活，随后凋亡和 ROS 产生增加，UCP2 表达降低。浓度为 0.75 和 1.5 mg/ml 的 YQHX 治疗显著降低了 Bax 和 caspase-3 的表达，而增加了 Bcl-2 和 UCP2 的蛋白表达。用 Lenti-shUCP2 载体转染敲低 UCP2 可减弱这些变化。
综上所述，本研究表明 YQHX 通过上调 H9c2 细胞中 UCP2 的表达来减轻 HO 诱导的细胞凋亡，提示 YQHX 是治疗 I/R 损伤介导的细胞凋亡的一种有前途的治疗方法。
