Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage.
Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage.
复制标题
DOI：
10.4162/nrp.2015.9.6.586
复制
发表时间：
2015-12
影响因子：
2.4
通讯作者：
Park WH
中科院分区：
文献类型：
--
作者：
Lee KP;Kim JE;Park WH
活性氧自由基(ROS)的形成与咪康唑所致的心功能障碍密切相关。虽然鼠李素具有抗氧化作用，但它是否能对抗咪康唑诱导的心肌细胞凋亡尚不清楚。因此，我们研究了鼠李素对咪康唑诱导的H9c2细胞凋亡的影响。用倒置显微镜观察细胞形态，用WelCount™细胞增殖分析试剂盒检测细胞活力。用6-羧基-2‘，7’-二氯二乙酸酯(H_2DCF-DA)染色荧光激活细胞分选法评价咪康唑诱导的ROS产生。免疫印迹法检测APE/Ref-1和半胱氨酸天冬氨酸蛋白酶(Caspase)3的表达。用实时定量聚合酶链式反应检测NADPH氧化酶水平。咪康唑(3和10µM)可引起H9c2细胞形态异常改变和细胞死亡。鼠李素以剂量依赖的方式提高咪康唑(3µM)处理的细胞存活率。鼠李素(1和3µM)可下调咪康唑刺激的细胞中caspase 3的裂解，上调APE/Ref-1的表达。此外，鼠李素显著减少了ROS的产生。我们的结果提示鼠李素可能通过抑制ROS对咪康唑刺激的H9c2心肌细胞具有细胞保护作用。这种作用很可能是通过上调APE/Ref-1和降低过氧化氢水平来实现的。
Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount™ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate (H2DCF-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. Miconazole (3 and 10 µM) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole (3 µM)-treated cells in a dose-dependent manner. Rhamnetin (1 and 3 µM) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.