Astragalus polysaccharides inhibit oxidation in high glucose-challenged or SOD2-silenced H9C2 cells.
Astragalus polysaccharides inhibit oxidation in high glucose-challenged or SOD2-silenced H9C2 cells.
复制标题
黄芪多糖抑制高葡萄糖挑战或 SOD2 沉默的 H9C2 细胞的氧化
DOI：
10.2147/dmso.s177269
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Yang Y
中科院分区：
文献类型：
--
作者：
Chen W;Sun Q;Ju J;Chen W;Zhao X;Zhang Y;Yang Y
前言氧化应激在糖尿病性心肌病（DCM）的发生发展中起着重要作用。以前，我们报道黄芪多糖（APS）通过抑制心肌氧化应激改善DCM。在这项研究中，我们评估了APS对高糖诱导的体外心肌细胞氧化应激的有益作用。材料与方法H9 C2细胞在高浓度葡萄糖的存在下培养或用siRNASOD 2转染，随后用APS处理。透射电镜观察细胞线粒体超微结构。使用发夹寡核苷酸探针检测细胞凋亡，并通过流式细胞术分析进行定量。使用荧光染料二氢乙锭（DHE）通过免疫组织化学测定超氧化物的产生。采用硝基酪氨酸和8-OH-dG抗体分别检测细胞质蛋白的氧化损伤和细胞核中的氧化应激。利用SOD检测试剂盒测定超氧化物歧化酶（SOD）活性，并通过Western印迹法分析SOD蛋白水平。结果黄芪多糖能保护高糖诱导和/或SOD 2沉默的H9 C2细胞线粒体超微结构，减少细胞凋亡（hairpin-1），抑制细胞内超氧化物产生（DHE），减少细胞质蛋白（硝基酪氨酸）氧化损伤和细胞核内氧化应激（8-OH-dG），并诱导SOD 2酶活性和蛋白水平升高。结论黄芪多糖对高糖刺激的H9 C2细胞具有保护作用，其机制可能与抑制氧化应激有关。
Introduction Oxidative stress plays an important role in the development of diabetic cardio-myopathy (DCM). Previously, we reported that Astragalus polysaccharides (APS) improved DCM by inhibition of cardiac oxidative stress. In this study, we evaluated the beneficial effect of APS on high glucose-induced oxidative stress in cardiomyocytes in vitro. Materials and methods H9C2 cells were cultured in the presence of high concentration of glucose or transfected with siRNASOD2, followed by APS treatment. The cellular mitochondrial ultrastructure was observed using a transmission electron microscope. Cell apoptosis was detected using hairpin oligonucleotide probes and quantified by flow cytometry analysis. Superoxide production was determined by immunohistochemistry using the fluorescent dye dihydroethidium (DHE). Nitrotyrosine and 8-OH-dG antibodies were employed to detect oxidative damage to cytoplasmic proteins and oxidative stress in the nuclei, respectively. Superoxide dismutase (SOD) activity was measured utilizing the SOD Assay Kit, and SOD protein levels were analyzed by Western blotting. Results APS treatment protected cellular mitochondrial ultrastructure, reduced cell apoptosis (hairpin-1), inhibited cellular superoxide production (DHE), and reduced oxidative damage to cytoplasmic proteins (nitrotyrosine) and oxidative stress in the nuclei (8-OH-dG) in high glucose-induced and/or SOD2-silenced H9C2 cells, together with induction of SOD2 enzyme activity and increase of protein levels. Conclusion Our findings indicated the beneficial effect of APS on high glucose-challenged H9C2 cells, which was associated with inhibition of oxidative stress in vitro.