论文部分内容阅读
【目的】探讨去琥珀酰化酶Sirtuin5(SIRT5)对受体相互作用蛋白140(RIP140)诱导的心肌线粒体能量代谢紊乱的影响。【方法】利用腺病毒感染心肌细胞诱导RIP140过表达,利用质粒转染诱导SIRT5基因过表达,si RNA干扰敲低RIP140、SIRT5表达;q RT-PCR和Western blot检测SIRT5、RIP140的表达变化;q RT-PCR检测线粒体编码基因的表达情况,TMRE染色法评估线粒体膜电位,试剂盒检测细胞耗氧和ATP生成情况。所有数据均来自至少3次的独立实验。【结果】过表达RIP140明显抑制SIRT5的表达(P<0.05);敲低内源性RIP140能增加SIRT5的表达(P<0.05)。过表达RIP140亦能诱导线粒体蛋白高度琥珀酰化,抑制SIRT5的去琥珀酰化酶活性。除此,RIP140能诱导线粒体功能紊乱和能量代谢损伤作用,表现为线粒体编码基因的表达抑制(P<0.05),线粒体膜电位降低(P<0.05),细胞耗氧和ATP合成减少(P<0.05),而过表达SIRT5能抑制RIP140介导的上述能量代谢损伤(P<0.05)。此外,SIRT5受RIP140的负性调控作用依赖于过氧化物增殖体激活受体α(PPARα)。【结论】RIP140通过抑制SIRT5诱导心肌细胞线粒体功能紊乱和能量代谢损伤。
【Aim】 To investigate the effect of Sirtuin5 (SIRT5) on disturbance of myocardial mitochondrial energy metabolism induced by receptor interacting protein 140 (RIP140). 【Methods】 The adenovirus-infected cardiomyocytes were used to induce overexpression of RIP140. SIRT5 overexpression was induced by plasmid transfection. The si RNA interference knockdown the expression of RIP140 and SIRT5. The expression of SIRT5 and RIP140 was detected by q RT-PCR and Western blotting. Q The expression of mitochondrial encoding gene was detected by RT-PCR, the mitochondrial membrane potential was evaluated by TMRE staining, and the cell oxygen consumption and ATP production were detected by kit. All data are from at least 3 independent experiments. [Results] Overexpression of RIP140 significantly inhibited SIRT5 expression (P <0.05). Knockdown of endogenous RIP140 increased SIRT5 expression (P <0.05). Overexpression of RIP140 also induced a high degree of succinylation of mitochondrial proteins and inhibited the desuccinylase activity of SIRT5. In addition, RIP140 could induce mitochondrial dysfunction and energy metabolism injury, which was inhibited by mitochondrial gene expression (P <0.05), decreased mitochondrial membrane potential (P <0.05) and decreased cellular oxygen consumption and ATP synthesis ), While overexpression of SIRT5 inhibited RIP140-mediated energy metabolism (P <0.05). In addition, SIRT5 is negatively regulated by RIP140 and is dependent on peroxisome proliferator activated receptor alpha (PPARα). 【Conclusion】 RIP140 can inhibit the mitochondrial dysfunction and energy metabolism of cardiomyocytes induced by SIRT5.