Regional Cardiovascular Disease Center, Department of Internal Medicine, Chungbuk National University School of Medicine, Cheongju, Korea.
Korean Circ J. 2012 Jan;42(1):23-32. doi: 10.4070/kcj.2012.42.1.23. Epub 2012 Jan 31.
The redox system is an important anti-oxidative system composed of thioredoxin, thioredoxin reductase, and peroxiredoxin (PRx). The fine details of PRx expression and its protective effects in various cells in cardiovascular tissue under oxidative stress created by hydrogen peroxide have not been fully elucidated.
Oxidative stress was induced by adding hydrogen peroxide at 0.25 mM for 2 hours to rat neonatal cardiomyocytes (rCMCs), rat vascular smooth muscle cells (rVSMCs), and human umbilical vein endothelial cells (HUVECs). Apoptosis was quantified by flow cytometry and the expression patterns of the six PRx isoforms were evaluated by western blotting in the three cell lines after hydrogen peroxide stimulation. Apoptosis and the cell survival signal pathway were evaluated by PRx1 gene delivery using lentiviral vector in hydrogen peroxide stimulated rCMCs versus green fluorescence protein gene delivery.
Hydrogen peroxide induced 25% apoptosis in rCMCs. Furthermore, the PRx1 and 5 isoforms were found to be overexpressed in hydrogen peroxide treated rCMCs, and PRx1 overexpression by gene delivery was found to reduce hydrogen peroxide induced rCMCs apoptosis significantly. In addition, this effect was found to originate from cell survival pathway modification.
Hydrogen peroxide induced significant oxidative stress in rCMCs, rVSMCs, and HUVECs, and PRx1 overexpression using a lentiviral vector system significantly reduced hydrogen peroxide induced rCMCs apoptosis by upregulation of cell survival signals and downregulation of apoptotic signals. These findings suggest that PRx1 could be used as a treatment strategy for myocardial salvage in conditions of oxidative stress.
氧化还原系统是由硫氧还蛋白、硫氧还蛋白还原酶和过氧化物酶（PRx）组成的重要抗氧化系统。过氧化物氢诱导的心血管组织中各种细胞的 PRx 表达及其在氧化应激下的保护作用的细节尚未完全阐明。
用 0.25mM 过氧化氢处理 2 小时诱导氧化应激，以建立大鼠新生心肌细胞（rCMCs）、大鼠血管平滑肌细胞（rVSMCs）和人脐静脉内皮细胞（HUVECs）的氧化应激模型。通过流式细胞术定量检测细胞凋亡，并用 Western blot 检测过氧化氢刺激后三种细胞系中 6 种 PRx 同工型的表达模式。用慢病毒载体传递 PRx1 基因，评估 PRx1 基因传递对过氧化氢刺激的 rCMCs 中细胞凋亡和细胞存活信号通路的影响，与绿色荧光蛋白基因传递进行比较。
过氧化氢诱导 rCMCs 凋亡 25%。此外，发现 PRx1 和 5 同工型在过氧化氢处理的 rCMCs 中过度表达，并且基因传递的 PRx1 过表达可显著减少过氧化氢诱导的 rCMCs 凋亡。此外，这种作用源自细胞存活途径的修饰。
过氧化氢在 rCMCs、rVSMCs 和 HUVECs 中诱导了明显的氧化应激，慢病毒载体系统中的 PRx1 过表达通过上调细胞存活信号和下调细胞凋亡信号，显著减少过氧化氢诱导的 rCMCs 凋亡。这些发现表明 PRx1 可作为氧化应激条件下心肌保护的治疗策略。
