Involvement of PINK1/Parkin-mediated mitophagy in AGE-induced cardiomyocyte aging.
Involvement of PINK1/Parkin-mediated mitophagy in AGE-induced cardiomyocyte aging.
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DOI：
10.1016/j.ijcard.2016.11.161
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发表时间：
2017-01
影响因子：
3.5
通讯作者：
Zhimin Zha;Junhong Wang;Xiangming Wang;Miao Lu;Yan Guo
中科院分区：
文献类型：
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作者：
Zhimin Zha;Junhong Wang;Xiangming Wang;Miao Lu;Yan Guo
背景和目的晚期糖基化终产物（AGEs）可诱导心肌细胞衰老。然而，其潜在的分子机制尚不清楚。方法用AGEs培养新生大鼠心肌细胞，通过衰老相关β-半乳糖苷酶（SA-β-gal）活性和衰老相关p16表达来评价细胞衰老情况。此外，通过测量PINK1、Parkin、LC3和p62蛋白的表达来评估有丝分裂活性。然后将线粒体自噬抑制剂环孢素A （cyclosporine A, CsA）或PINK1 sirna给予心肌细胞，研究线粒体自噬在age诱导的衰老中的作用。结果经AGEs处理的心肌细胞SA-β-gal阳性细胞数量明显增加，p16蛋白水平明显升高。此外，AGEs显著提高了PINK1和Parkin蛋白水平以及LC3-II/LC3-I比值，且呈剂量依赖性。然而，与对照组相比，AGE组p62的表达明显下降。令人惊讶的是，CsA和小干扰RNA （siRNA）敲低PINK1均显著降低了age处理心肌细胞的LC3-II/LC3-I比率以及PINK1和Parkin蛋白水平。此外，CsA处理或敲低PINK1表达减弱了AGEs诱导的心肌细胞中SA-β-gal阳性细胞数量的增加和p16水平的上调。结论spink1 / parkinson介导的线粒体自噬参与了AGEs诱导的心肌细胞衰老过程，降低线粒体自噬活性可能是阻断心肌细胞衰老的有效途径。
Context and objectivesAdvanced glycation end products (AGEs) can induce senescence in cardiomyocytes. However, its underlying molecular mechanisms remain unknown.MethodsNeonatal rat cardiomyocytes were incubated with AGEs, and cellular senescence was evaluated by senescence-associated beta-galactosidase (SA-β-gal) activity and aging-associated p16 expression. In addition, mitophagic activity was evaluated by measuring the expression of the PINK1, Parkin, LC3 and p62 proteins. The mitophagy inhibitor cyclosporine A (CsA) or PINK1 siRNAs was then administered to cardiomyocytes to study the role of mitophagy in AGE-induced aging.ResultsA significantly increased number of SA-β-gal positive cells and increased p16 protein levels were observed in cardiomyocytes treated with AGEs. Moreover, AGEs significantly increased the protein levels of PINK1 and Parkin as well as the LC3-II/LC3-I ratio, which occurred in a dose-dependent manner. However, the expression of p62 decreased significantly in the AGE group compared to the control. Surprisingly, both CsA and the knockdown of PINK1 by small-interfering RNA (siRNA) significantly decreased the LC3-II/LC3-I ratio and the PINK1 and Parkin protein levels in AGE-treated cardiomyocytes. Moreover, CsA treatment or knockdown of PINK1 expression attenuated the increased number of SA-β-gal positive cells and the upregulated p16 level in cardiomyocytes induced by AGEs.ConclusionsPINK1/Parkin-mediated mitophagy is involved in the process of cardiomyocyte senescence induced by AGEs, and a reduction in mitophagic activity might be a promising approach to block the senescent state in cardiomyocytes.