芒果苷减轻缺氧复氧所致人心肌细胞损伤

目的:探讨芒果苷减轻缺氧复氧所致人心肌细胞损伤的效应及机制。方法:将人心肌AC16细胞分为正常对照组、缺氧复氧组及芒果苷(50、100和200μmol/L)+缺氧复氧组。分别采用RT-qPCR及Western blot法检测Kelch样环氧氯丙烷相关蛋白1(Keap-1)、Bax、Bcl-2、caspase-3、caspase-9和超氧化物歧化酶2(SOD2)的mRNA及蛋白表达;Western blot法检测细胞核中核因子E2相关因子2(Nrf-2)的表达水平;RT-qPCR法检测miR-432-3p的表达;DCFH-DA探针检测细胞内活性氧簇(ROS)的生成;CCK-8法检测细胞活力;流式细胞术检测细胞凋亡。结果:相对于正常对照组,缺氧复氧处理AC16细胞后,miR-432-3p和Keap-1的mRNA及蛋白表达无明显变化,Nrf-2核转位和ROS生成增多(P<0. 05),Bax、caspase-3和caspase-9的mRNA及蛋白水平显著升高(P<0. 05),SOD2和Bcl-2的mRNA及蛋白水平以及细胞活力显著降低(P<0. 05),细胞凋亡显著增多(...

Mangiferin attenuates hypoxia/reoxygenation-induced injury of human myocardial cells

AIM:To investigate the effect of mangiferin on hypoxia/reoxygenation(H/R)-induced injury of human myocardial cells and its mechanism.METHODS:Human myocardial AC16 cells were divided into normal group,H/R group and H/R+mangiferin(50,100 and 200μmol/L)treatment groups.The mRNA and protein expression levels of Kelch-like epichlorohydrin-associated protein-1(Keap-1),Bax,Bcl-2,caspase-3,caspase-9 and superoxide dismutase 2(SOD2)were detected by RT-qPCR and Western blot,respectively.The protein expression of nuclear factor E2-related factor 2(Nrf-2)in nucleus was determined by Western blot.The expression of microRNA-432-3p(miR-432-3 p)was detected by RT-qPCR.The generation of reactive oxygen speciess(ROS)in the cells was measured by DCFH-DA probing.The cell viability was measured by CCK-8 assay.Apoptosis was analyzed by flow cytometry.RESULTS:No significant difference in the expression of miR-432-3p and Keap-1 between normal group and H/R group was observed.Compared with normal group,the nuclear translocation of Nrf-2,the ROS level,and the mRNA and protein expression of Bax,caspase-3 and caspase-9 were significantly increased in H/R group(P<0.05).The mRNA and protein expression of SOD2 and Bcl-2,and the cell viability significantly decreased in H/R group compared with normal group,while the apoptosis was increased significantly(P<0.05).Treatment with mangiferin resulted in an increase in the miR-432-3p expression and a decrease in the ROS level,and the expression of Keap-1,Bax,caspase-3 and caspase-9 was also inhibited as compared with H/R group(P<0.05).The Nrf-2 nuclear translocation,and the protein levels of SOD2 and Bcl-2 in mangiferin treatment groups were significantly increased as compared with H/R group(P<0.05).The cell viability was increased significantly,and the apoptosis was decreased significantly in mangiferin treatment groups as compared with H/R group(P<0.05).The effects of mangiferin in middle-and high-dose groups were better than those in low-dose group,and no significant difference between middle-and high-dose groups was found.CONCLUSION:Mangiferin inhibits the decrease in myocardial cell viability and the apoptosis induced by H/R injury.The mechanism may be related to the up-regulation of Nrf-2 antioxidant stress effect via enhancing the expression of miR-432-3p.