芒果苷对心肌细胞损伤的保护作用及对NFATc4表达的影响
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篇名: 芒果苷对心肌细胞损伤的保护作用及对NFATc4表达的影响
TITLE: Protective effects of mangiferin against myocardial cell damage and its effects on the expression of NFATc4
摘要: 目的 研究芒果苷对过氧化氢(H2O2)诱导的心肌细胞氧化应激损伤的保护作用,以及对活化T细胞核因子c4(NFATc4)表达的影响。方法体外培养心肌H9c2细胞,分为空白组、H2O2组和芒果苷50、100、150μmol/L组,芒果苷组细胞在经不同浓度芒果苷作用12h后,再与H2O2组一同经H2O(2200μmol/L)刺激12h,检测各组细胞的相对存活率,细胞上清液中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)水平,细胞中活性氧(ROS)水平,以及细胞中凋亡相关蛋白[B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、剪切的胱天蛋白酶3(cleavedcaspase-3)]、核蛋白NFATc4的表达情况。转染NFATc4干扰序列,考察NFATc4对H2O2诱导的心肌细胞中氧化应激指标和凋亡相关蛋白的影响。结果与空白组比较,H2O2组细胞的相对存活率,SOD、CAT水平,Bcl-2的相对表达量均显著降低;MDA、ROS水平,Bax、cleavedcaspase-3和核蛋白NFATc4的相对表达量均显著升高(P<0.05或P<0.01)。与H2O2组比较,芒果苷100、150μmol/L组细胞的上述指标均得以显著逆转(P<0.05)。转染NFATc4干扰序列后,核蛋白NFATc4的表达显著下调,MDA、ROS水平和Bax、cleavedcaspase-3蛋白的表达均较H2O2组显著降低/下调,SOD、CAT水平和Bcl-2蛋白的表达均较H2O2组显著升高/上调(P<0.05)。结论芒果苷能够减轻H2O2诱导的H9c2细胞氧化应激,减少细胞凋亡,抑制NFATc4蛋白入核,进而缓解心肌细胞损伤;降低NFATc4蛋白的核内水平与减轻H2O2诱导的氧化应激、细胞凋亡有关。
ABSTRACT: OBJECTIVE To study the protective effects of mangiferin against oxidative stress injury of myocardial cells induced by hydrogen peroxide (H2O2), and its effects on the expression of nuclear factor of activated T cell cytoplasmic 4(NFATc4). METHODS H9c2 myocardial cells were cultured in vitro and divided into blank group, H2O2 group, and 50, 100, 150 μmol/L mangiferin groups. Mangiferin groups were treated with different concentrations of mangiferin for 12 h, and then were subjected to H2O2 (200 μmol/L) stimulation for 12 hours together with the H2O2 group; relative survival rate was detected in each group, and the levels of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in cell supernatant and reactive oxygen species (ROS) in H9c2 cells were measured. Meanwhile, the expressions of apoptosis-related proteins [B cell lymphoma 2 (Bcl- 2), Bcl-2 associated X protein (Bax), cleaved caspase-3] and nuclear protein NFATc4 were determined. Furthermore, the NFATc4 interference sequence was transfected, and the effects of NFATc4 on oxidant stress indexes and apoptosis-related proteins in H2O2- induced myocardial cells were investigated. RESULTS Compared with blank group, relative cell viability, the levels of SOD and CAT, relative expression of Bcl-2 were decreased significantly, while the levels of MDA and ROS, relative expressions of Bax, cleaved caspase-3 and nuclear protein NFATc4 were increased significantly (P<0.05 or P<0.01). Compared with the H2O2 group, the above indexes of 100 and 150 μmol/L mangiferin groups were reversed significantly (P<0.05). After the transfection of the NFATc4 interference sequence, the expression of nuclear protein NFATc4 was down-regulated significantly; the levels of MDA and SOD, the protein expressions of Bax and cleaved caspase-3 were all decreased/down-regulated significantly, while the levels of SOD and CAT, and the protein expression of Bcl-2 were all increased/up-regulated significantly, compared with H2O2 group (P< 0.05). CONCLUSIONS Mangiferin can relieve H2O2-induced oxidative stress of H9c2 cells, reduce the apoptosis and inhibit the nuclear translocation of NFATc4, thereby alleviating myocardial cell damage; reducing the nuclear level of NFATc4 protein is related to reducing H2O2-induced oxidative stress and cell apoptosis.
期刊: 2023年第34卷第20期
作者: 韦啟球;高思;冯艺萍;裴世成;刘雪萍
AUTHORS: WEI Qiqiu,GAO Si,FENG Yiping,PEI Shicheng,LIU Xueping
关键字: 芒果苷;H9c2细胞;氧化应激;凋亡;活化T细胞核因子c4
KEYWORDS: mangiferin; H9c2 cells; oxidative stress;
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