Department of Environmental Health Sciences, University of Alabama at Birmingham, Birmingham, Alabama;
Am J Physiol Gastrointest Liver Physiol. 2014 Feb 15;306(4):G265-77. doi: 10.1152/ajpgi.00278.2013. Epub 2013 Dec 19.
Chronic ethanol consumption increases sensitivity of the mitochondrial permeability transition (MPT) pore induction in liver. Ca(2+) promotes MPT pore opening, and genetic ablation of cyclophilin D (CypD) increases the Ca(2+) threshold for the MPT. We used wild-type (WT) and CypD-null (CypD(-/-)) mice fed a control or an ethanol-containing diet to investigate the role of the MPT in ethanol-mediated liver injury. Ca(2+)-mediated induction of the MPT and mitochondrial respiration were measured in isolated liver mitochondria. Steatosis was present in WT and CypD(-/-) mice fed ethanol and accompanied by increased terminal deoxynucleotidyl transferase dUTP-mediated nick-end label-positive nuclei. Autophagy was increased in ethanol-fed WT mice compared with ethanol-fed CypD(-/-) mice, as reflected by an increase in the ratio of microtubule protein 1 light chain 3B II to microtubule protein 1 light chain 3B I. Higher levels of p62 were measured in CypD(-/-) than WT mice. Ethanol decreased mitochondrial respiratory control ratios and select complex activities in WT and CypD(-/-) mice. Ethanol also increased CypD protein in liver of WT mice. Mitochondria from control- and ethanol-fed WT mice were more sensitive to Ca(2+)-mediated MPT pore induction than mitochondria from their CypD(-/-) counterparts. Mitochondria from ethanol-fed CypD(-/-) mice were also more sensitive to Ca(2+)-induced swelling than mitochondria from control-fed CypD(-/-) mice but were less sensitive than mitochondria from ethanol-fed WT mice. In summary, CypD deficiency was associated with impaired autophagy and did not prevent ethanol-mediated steatosis. Furthermore, increased MPT sensitivity was observed in mitochondria from ethanol-fed WT and CypD(-/-) mice. We conclude that chronic ethanol consumption likely lowers the threshold for CypD-regulated and -independent characteristics of the ethanol-mediated MPT pore in liver mitochondria.
慢性乙醇摄入增加了肝脏线粒体通透性转换孔(MPT)诱导的敏感性。钙离子促进 MPT 孔开放,环孢素 D(CypD)的基因缺失增加了 MPT 的钙离子阈值。我们使用野生型(WT)和 CypD 缺失(CypD(-/-))小鼠,给予对照或含乙醇饮食,以研究 MPT 在乙醇介导的肝损伤中的作用。在分离的肝线粒体中测量钙介导的 MPT 和线粒体呼吸的诱导。WT 和 CypD(-/-) 小鼠在给予乙醇时存在脂肪变性,并伴有末端脱氧核苷酸转移酶 dUTP 介导的切口末端标记阳性核增加。与 CypD(-/-) 相比,乙醇喂养的 WT 小鼠中的自噬增加,这反映在微管蛋白 1 轻链 3B II 与微管蛋白 1 轻链 3B I 的比值增加。在 CypD(-/-) 中测量到的 p62 水平高于 WT 小鼠。乙醇降低了 WT 和 CypD(-/-) 小鼠线粒体呼吸控制比和选择复合物活性。乙醇还增加了 WT 小鼠肝脏中的 CypD 蛋白。与 CypD(-/-) 相比,来自对照和乙醇喂养 WT 小鼠的线粒体对钙离子介导的 MPT 孔诱导更为敏感。来自乙醇喂养 CypD(-/-) 小鼠的线粒体对钙离子诱导的肿胀也比来自对照喂养 CypD(-/-) 小鼠的线粒体更为敏感,但比来自乙醇喂养 WT 小鼠的线粒体更为敏感。总之,CypD 缺失与自噬受损有关,并且不能预防乙醇介导的脂肪变性。此外,在来自乙醇喂养 WT 和 CypD(-/-) 小鼠的线粒体中观察到 MPT 敏感性增加。我们得出结论,慢性乙醇摄入可能会降低 CypD 调节的和独立的乙醇介导的 MPT 孔在肝线粒体中的阈值。