【摘要】 目的:研究乌梅有机酸对缺氧/复氧(H/R)诱导心肌细胞氧化应激损伤的影响及其作用机制。方法:将心肌细胞H9C2随机分为对照组、H/R组、H/R+不同浓度乌梅有机酸组。MTT法检测心肌细胞存活,试剂盒检测超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性、丙二醛(MDA)水平、天冬氨酸转氨酶(AST)水平、肌酸激酶(CK)水平、乳酸脱氢酶(LDH)水平,流式细胞术检测心肌细胞凋亡,Western blotting检测Bcl2相关X蛋白(Bax)、B细胞淋巴瘤/白血病2(Bcl2)、核转录因子E2相关因子2(Nrf2)-抗氧化反应元件(ARE)信号通路相关蛋白Nrf2、血红素加氧酶1(HO-1)和醌氧化还原酶1(NQO1)的表达。结果:与对照组相比,H/R组心肌细胞存活率、SOD活性、CAT活性显著提高,MDA水平、AST水平、CK水平和LDH水平明显降低,且细胞凋亡率、Bax蛋白表达量增加,Bcl2、Nrf2、HO-1、NQO1蛋白表达量减少,差异均有统计学意义(P <0.05)。与H/R组比较,H/R+1、2 mg·ml-1乌梅有机酸组H/R损伤心肌细胞的存活率明显提高(均P <0.05); H/R损伤心肌细胞的SOD活性、CAT活性显著增加,MDA水平、AST水平、CK水平和LDH水平降低,以及心肌细胞凋亡率、Bax蛋白表达量减少,Bcl2、Nrf2、HO-1、NQO1蛋白水平提高,差异均有统计学意义(P <0.05)。结论:乌梅有机酸可能通过激活Nrf2-ARE信号通路保护H/R导致的心肌细胞氧化应激损伤。更多还原

【Abstract】 Objective: To study the effect of organic acid from Fructus mume on oxidative stress injury induced by hypoxia/reoxygenation( H/R) in cardiomyocytes and its mechanism. Methods: Myocardial cells H9 C2 were randomly divided into control group,H/R group,H/R + group of different concentrations of organic acid.Myocardial cell survival was detected by MTT assay. Superoxide dismutase( SOD) activity,catalase( CAT)activity,malondialdehyde( MDA) content,aspartate aminotransferase( AST) level,creatine kinase( CK) level and lactate dehydrogenase( LDH) level were determined by kit. Apoptosis of cardiomyocytes was detected by flow cytometry. The expressions of Bax,Bcl2 and Nrf2-ARE signaling pathway-related proteins Nrf2,HO-1 and NQO1 were analyzed by Western blotting. Results: Compared with the control group,H/R treatment significantly reduced myocardial cell survival rate,SOD activity,CAT activity,obviously improved MDA content,AST,CK and LDH levels,and increased apoptosis rate and Bax protein expression,as well as reduced the expression of Bcl2,Nrf2,HO-1,NQO1 protein,and the differences were all significant( P < 0. 05). Compared with H/R group,1,2 mg·ml-1 organic acid from Fructus mume significantly increased the survival rate of H/R injured cardiomyocytes( P <0. 05). Compared with H/R group,organic acid from Fructus mume significantly increased SOD activity and CAT activity of H/R injured cardiomyocytes,decreased MDA content,AST,CK and LDH levels,myocardial apoptosis rate and Bax protein expression,increased Bcl2,Nrf2,HO-1,NQO1 protein levels,and the differences were all significant( P < 0. 05). Conclusion: Organic acid from Fructus mume protects oxidative stress damage induced by H/R by activating Nrf2-ARE signaling pathway.更多还原