Induced pluripotent stem cell-conditional medium inhibits H9C2 cardiomyocytes apoptosis via autophagy flux and Wnt/β-catenin pathway
Induced pluripotent stem cell-conditional medium inhibits H9C2 cardiomyocytes apoptosis via autophagy flux and Wnt/β-catenin pathway
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诱导多能干细胞条件培养基通过自噬流和 Wnt/β-连环蛋白途径抑制 H9C2 心肌细胞凋亡
DOI：
10.1111/jcmm.14327
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发表时间：
2019-06-01
影响因子：
5.3
通讯作者：
Chu, Maoping
中科院分区：
文献类型：
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作者：
Guo, Xiaoling;Gu, Xiaohong;Chu, Maoping
诱导多能干细胞来源的条件培养基（iPS-CM）可以提高多种类型细胞的活力，可能是治疗心肌梗塞的更好替代方案。本研究旨在探讨iPS-CM对H9C2心肌细胞抗凋亡和增殖的影响并探讨其潜在机制。将 H9C2 心肌细胞暴露于 200 μmol/L 过氧化氢 (H2O2) 中 24 小时，无论是否经过 iPS-CM 预处理。通过流式细胞仪分析凋亡细胞的比例、线粒体膜电位的损失（Delta Psi m）和细胞内活性氧的水平。通过Western blot分析BCL-2和BAX蛋白的表达水平。使用细胞周期和 EdU 染色测定评估细胞增殖。为了研究细胞衰老，进行了衰老相关的β-半乳糖苷酶（SA-β-gal）染色。丙二醛、超氧化物歧化酶和谷胱甘肽的水平也使用市售酶试剂盒进行定量。结果表明，含有碱性成纤维细胞生长因子的iPS-CM通过激活自噬流通路显着减少H2O2诱导的H9C2心肌细胞凋亡，通过上调Wnt/β-catenin通路促进心肌细胞增殖，并抑制氧化应激和细胞衰老。总之，iPS-CM有效增强了H9C2心肌细胞的细胞活力，未来有望用于抑制心肌细胞凋亡以治疗心肌梗死。
Induced pluripotent stem cell-derived conditioned medium (iPS-CM) could improve cell viability in many types of cells and may be a better alternative for the treatment of myocardial infarction. This study aimed to examine the influence of iPS-CM on anti-apoptosis and the proliferation of H9C2 cardiomyocytes and investigate the underlying mechanisms. H9C2 cardiomyocytes were exposed to 200 mu mol/L hydrogen peroxide (H2O2) for 24 hours with or without pre-treatment with iPS-CM. The ratio of apoptotic cells, the loss of mitochondrial membrane potential (Delta Psi m) and the levels of intracellular reactive oxygen species were analysed by flow cytometric analysis. The expression levels of BCL-2 and BAX proteins were analysed by Western blot. Cell proliferation was assessed using cell cycle and EdU staining assays. To study cell senescence, senescence-associated beta-galactosidase (SA-beta-gal) staining was conducted. The levels of malondialdehyde, superoxide dismutase and glutathione were also quantified using commercially available enzymatic kits. The results showed that iPS-CM containing basic fibroblast growth factor significantly reduced H2O2-induced H9C2 cardiomyocyte apoptosis by activating the autophagy flux pathway, promoted cardiomyocyte proliferation by up-regulating the Wnt/beta-catenin pathway and inhibited oxidative stress and cell senescence. In conclusion, iPS-CM effectively enhanced the cell viability of H9C2 cardiomyocytes and could potentially be used to inhibit cardiomyocytes apoptosis to treat myocardial infarction in the future.