Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy
Ethanol-Induced Mitochondrial Damage in Sertoli Cells is Associated with Parkin Overexpression and Activation of Mitophagy
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DOI：
10.3390/cells8030283
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发表时间：
2019-03-25
期刊：
影响因子：
6
通讯作者：
Kondo, Yoichi
中科院分区：
文献类型：
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作者：
Eid, Nabil;Ito, Yuko;Kondo, Yoichi
本研究旨在阐明PINK 1-Parkin通路参与乙醇诱导的Sertoli细胞（SC）线粒体自噬。在研究中，成年大鼠腹腔注射乙醇（5 gm/kg），并在24 h内的不同时间段处死。透射电子显微镜显示，乙醇处理的大鼠（ETRs）的SC中线粒体损伤增强，与磷酸盐缓冲盐水（PBS）处理的对照组中线粒体吞噬空泡（线粒体吞噬体和自体溶酶体）数量显著增加相比，线粒体损伤增强。这种增强通过观察LC 3标记的膜内的捕获的线粒体、LC 3蛋白水平的上调、LC 3和细胞色素c的共定位以及线粒体蛋白的表达减少而在超微结构上得到验证。重要的是，发现帕金表达在ETR SC中上调，特别是在线粒体和线粒体吞噬体中，除了与PINK 1和泛组织蛋白酶共定位之外，表明线粒体吞噬增强。转录因子EB（TFEB，一种自噬和线粒体自噬蛋白的转录因子）也被发现在ETR SC的细胞核中上调，并与iNOS的表达增强相关。在ETR SC中增强的Parkin相关的线粒体自噬可能是具有治疗意义的保护机制。据作者所知，这是第一个报告，证明了帕金森氏症相关的线粒体自噬在ETR干细胞的超微结构特征和分子机制。
This study was conducted to elucidate the involvement of the PINK1-Parkin pathway in ethanol-induced mitophagy among Sertoli cells (SCs). In the research, adult rats were given intraperitoneal injections of ethanol (5 gm/kg) and sacrificed at various time periods within 24 h. Transmission electron microscopy was applied to reveal enhanced mitochondrial damage in SCs of the ethanol-treated rats (ETRs) in association with a significant increase in numbers of mitophagic vacuoles (mitophagosomes and autolysosomes) in contrast to very low levels in a control group treated with phosphate-buffered saline (PBS). This enhancement was ultra-structurally verified via observation of trapped mitochondria within LC3-labeled membranes, upregulation of LC3 protein levels, colocalization of LC3 and cytochrome c, and reduced expression of mitochondrial proteins. Importantly, Parkin expression was found to be upregulated in ETR SCs, specifically in mitochondria and mitophagosomes in addition to colocalization with PINK1 and pan-cathepsin, indicating augmented mitophagy. Transcription factor EB (TFEB, a transcription factor for autophagy and mitophagy proteins) was also found to be upregulated in nuclei of ETR SCs and associated with enhanced expression of iNOS. Enhanced Parkin-related mitophagy in ETR SCs may be a protective mechanism with therapeutic implications. To the authors' knowledge, this is the first report demonstrating the ultrastructural characteristics and molecular mechanisms of Parkin-related mitophagy in ETR SCs.