OBJECTIVE: To assess the function of Dex in mitochondrial dysfunction during MIRI.
METHODS: To investigate the function of Dex in MIRI, H9C2 cells were placed in condition of hypoxia/reoxygenation (H/R). CCK8 assay was performed to test the cell viability, and the mitochondrial membrane potential was evaluated by JC-1 staining. In addition, the binding relationship between Sirt3 and Prdx3 was explored by Co-IP assay. Furthermore, the protein expressions were examined using western blot.
RESULTS: Dex could abolish H/R-induced mitochondrial dysfunction in H9C2 cells. In addition, H/R treatment significantly inhibited the expression of Sirt3, while Dex partially restored this phenomenon. Knockdown of Sirt3 or Prdx3 obviously reduced the protective effect of Dex on H/R-induced mitochondrial injury. Meanwhile, Sirt3 could enhance the function of Prdx3 via deacetylation of Prdx3.
CONCLUSIONS: Dex was found to attenuate H/R-induced mitochondrial dysfunction in cardiomyocytes via activation of Sirt3/Prdx3 pathway. Thus, this study might shed new lights on exploring new strategies for the treatment of MIRI.
目的:评估Dex在MIRI期间线粒体功能障碍中的功能。
方法:为了研究Dex在MIRI中的功能,将H9C2细胞置于缺氧/复氧(H/R)条件下。进行CCK8测定以测试细胞活力,并通过JC-1染色评估线粒体膜电位。此外,通过Co-IP分析探索Sirt3和Prdx3之间的结合关系。此外,蛋白质表达采用蛋白质印迹法检测。
结果:Dex可以消除H/R诱导的H9C2细胞线粒体功能障碍。此外,H/R处理显著抑制了Sirt3的表达,而Dex部分恢复了这一现象。敲除Sirt3或Prdx3可明显降低Dex对H/R诱导的线粒体损伤的保护作用。同时,Sirt3可通过Prdx3的脱乙酰作用增强Prdx3的功能。
结论:发现Dex通过激活Sirt3/Prdx3通路减轻H/R诱导的心肌细胞线粒体功能障碍。因此,这项研究可能为探索MIRI治疗的新策略提供新的思路.