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本研究旨在探讨原花青素(proanthocyanidin,Pro)抗H9C2细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的机制,明确蛋白质酪氨酸激酶2/信号传导子与激活子3(Janus kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)信号通路在此过程中的作用。取对数生长期的H9C2细胞株,随机分为5组:对照组(Con)、H/R损伤组(H/R)、Pro处理组(H/R+Pro)、JAK2小干扰RNA处理组(H/R+Pro+JAK2 si RNA)和JAK2小干扰RNA对照组(H/R+JAK2 si RNA)。Pro(40μmol/L)预处理8 h,H9C2细胞系缺氧2 h、复氧4 h后用MTT和TUNEL法分别检测各组细胞活力和凋亡率,用试剂盒检测超氧化物生成量,用Western blot法检测JAK2/STAT3通路相关分子,氧化应激指标及内质网应激相关蛋白的表达。结果显示,与H/R组相比,Pro处理可显著提高H/R处理的H9C2细胞活力,并降低细胞凋亡率,明显上调p-JAK2及p-STAT3水平,下调氧化应激指标超氧化物生成量及gp91phox蛋白表达量,下调内质网应激标志蛋白葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)、CCAAT/增强子结合蛋白(CCAAT/enhancer binding protein homologous protein,CHOP)及caspase-12的表达,而Pro以上保护作用均被JAK2 si RNA所抑制。本研究结果表明,Pro可通过JAK2/STAT3信号通路减轻H/R引起的H9C2细胞氧化应激与内质网应激损伤。本研究为阐明Pro的心血管保护作用及相关药物的开发提供了实验依据。
This study was designed to investigate the mechanism of proanthocyanidin (Pro) against H9R2 cells hypoxia / reoxygenation (H / R) injury and to clarify the role of prokaryotic protein tyrosine kinase 2 / Janus kinase 2 / signal transducer and activator of transcription 3, JAK2 / STAT3) signaling pathways in this process. The H9C2 cell line in logarithmic growth phase was randomly divided into 5 groups: control group (H / R), H / R injury group, H / R + Pro group, JAK2 small interfering RNA treatment group (H / R + Pro + JAK2 si RNA) and JAK2 small interfering RNA control (H / R + JAK2 si RNA). Pro (40μmol / L) for 8 h. H9C2 cells were exposed to hypoxia for 2 h. After reoxygenation for 4 h, the cell viability and apoptosis rate were determined by MTT and TUNEL. The superoxide production was detected by kit, Western blot was used to detect JAK2 / STAT3 pathway related molecules, oxidative stress and endoplasmic reticulum stress-related protein expression. The results showed that compared with H / R group, Pro treatment significantly increased the viability of H9C2 cells treated with H / R, decreased the apoptosis rate, significantly increased the levels of p-JAK2 and p-STAT3, and decreased the levels of oxidative stress And the expression of gp91phox protein, downregulated the expression of endoplasmic reticulum stress-regulated protein 78 (GRP78), CCAAT / enhancer binding protein homologous protein (CHOP) and caspase -12 expression, while the above Pro protective effect by JAK2 si RNA inhibition. Our results show that Pro can reduce H / R induced H9C2 cell oxidative stress and endoplasmic reticulum stress injury through JAK2 / STAT3 signaling pathway. This study provides experimental evidence for elucidating the cardiovascular protective effect of Pro and the development of related drugs.