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目的探讨活性氧(ROS)激活的环氧合酶-2(COX-2)通路在高糖(HG)损伤H9c2心肌细胞中的作用及血管紧张素-(1-7)[Ang-(1-7)]能否通过调控ROS激活的COX-2通路抑制HG引起的心肌细胞损伤。方法应用Western blot法检测COX-2蛋白的表达;细胞计数盒(CCK-8)测定细胞存活率;Hoechst 33258核染色检测凋亡细胞的形态及数量的变化;DCFH-DA染色荧光显微镜测定细胞内ROS水平;JC-1染色荧光显微镜照相测定线粒体膜电位(MMP)。结果应用1000μmol/L ROS清除剂(N-乙酰半胱氨酸,NAC)或1μmol/L Ang-(1-7)共处理H9c2心肌细胞12 h能显著地抑制HG对COX-2表达的上调作用;35 mmol/L葡萄糖(HG)处理心肌细胞24 h引起明显的损伤作用,使细胞存活率和MMP降低,凋亡细胞数量及细胞内ROS生成增多;Ang-(1-7)或COX-2抑制剂(NS-398)共处理心肌细胞明显地抑制上述HG引起的损伤作用。结论 Ang-(1-7)通过抑制ROS激活COX-2通路保护心肌细胞对抗HG引起的损伤。
Objective To investigate the effect of cyclooxygenase-2 (COX-2) pathway activated by reactive oxygen species (ROS) on H9c2 cardiomyocytes injured by high glucose (HG) and the effect of angiotensin- (1-7) 7)] can inhibit HG-induced cardiomyocyte injury by regulating ROS-activated COX-2 pathway. Methods The expression of COX-2 protein was detected by Western blot. The cell viability was measured by CCK-8. The morphology and number of apoptotic cells were detected by Hoechst 33258 nuclear staining. The expression of COX- ROS level; JC-1 stained fluorescence microscopy measured mitochondrial membrane potential (MMP). Results Co-treatment of H9c2 cardiomyocytes with 1000μmol / L ROS scavenger (NAC, NAC) or 1μmol / L Ang- (1-7) for 12 h significantly inhibited the upregulation of COX-2 by HG ; 35 mmol / L glucose (HG) treatment of myocardial cells 24 h caused a significant injury, the cell survival rate and MMP decreased, the number of apoptotic cells and intracellular ROS production increased; Ang- (1-7) or COX-2 Co-treatment of cardiomyocytes with inhibitors (NS-398) significantly inhibited the above HG-induced injury. Conclusions Ang- (1-7) protects cardiomyocytes against HG-induced injury by inhibiting ROS-activated COX-2 pathway.