热休克转录因子1在AngⅡ诱导血管内皮细胞损伤和内质网应激中的作用

Role of heat shock transcription factor 1 in angiotensin II-induced endothelial cell injury and endoplasmic reticulum stress

目的:研究热休克转录因子1(HSF1)在血管紧张素Ⅱ(AngⅡ)诱导血管内皮细胞损伤和内质网应激中的作用。方法:通过转染pcDNA3.1-HSF1上调血管内皮细胞中HSF1表达,再用AngⅡ处理血管内皮细胞。二硝基苯肼比色法检测培养液中乳酸脱氢酶(LDH)水平;硝酸还原酶法检测细胞分泌一氧化氮(NO)水平;ELISA法检测细胞分泌的内皮素1(ET-1)水平;流式细胞术检测细胞凋亡;ELISA法检测细胞分泌的肿瘤坏死因子α(TNF-α)、白细胞介素 6(IL-6)和白细胞介素 1β(IL-1β)水平;Western blot检测细胞中CCAAT/增强子结合蛋白同源蛋白(CHOP)、活化转录因子6(ATF6)和活化型caspase-3(cleaved caspase-3)的蛋白水平。结果:高表达HSF1可以降低AngⅡ诱导下血管内皮细胞培养液中LDH水平,升高NO水平,降低ET-1水平,降低细胞凋亡率及细胞中CHOP、ATF6和cleaved caspase-3的蛋白水平,减少细胞分泌TNF-α、IL-6和IL-1β。结论:高表达HSF1减轻AngⅡ诱导的血管内皮细胞损伤和内质网应激,减少细胞分泌炎症因子。

AIM: To study the role of heat shock transcription factor 1 (HSF1) in angiotensin II (Ang II)-induced endothelial cell injury and endoplasmic reticulum stress. METHODS: Vascular endothelial cells were treated with Ang II. The level of lactate dehydrogenase (LDH) in the culture medium was measured by the dinitro-phenylhydrazine method. The level of nitric oxide (NO) secreted from the cells was measured by nitrate reductase assay. The levels of endothelin-1 (ET-1), tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and IL-1β secreted from the cells were measured by ELISA. The apoptosis was analyzed by flow cytometry. The protein levels of CCAAT/enhancer-binding protein homologous protein (CHOP), activating transcription factor 6 (ATF6) and cleaved caspase-3 in the cells were determined by Western blot. RESULTS: Over-expression of HSF1 decreased LDH level in culture medium of vascular endothelial cells with Ang II stimulation, increased NO level, reduced the level of ET-1, decreased the apoptotic rate and the protein levels of CHOP, ATF6 and cleaved caspase-3, and reduced the secretion of TNF-α, IL-6 and IL-1β by the cells. CONCLUSION: Over-expression of HSF1 attenuates endothelial cell injury and endoplasmic reticulum stress induced by Ang II, and reduces the secretion of inflammatory factors in the endothelial cells.