Cui Ruibing, Yan Lihui, Luo Zheng, Guo Xiaolan, Yan Ming
Department of Hepatology and Gastroenterology, Qilu Hospital of Shandong University, Jinan, Shandong Province 250012, PR China.
Shandong Normal University, Jinan, Shandong Province 250012, PR China.
Toxicol Appl Pharmacol. 2015 Aug 15;287(1):52-66. doi: 10.1016/j.taap.2015.05.014. Epub 2015 May 30.
Extracellular Ca(2+) influx has been suggested to play a role in ethanol-induced hepatocyte apoptosis and necrosis. Previous studies indicated that store-operated Ca(2+) entry (SOCE) was involved in liver injury induced by ethanol in HepG2 cells. However, the mechanisms underlying liver injury caused by SOCE remain unclear. We aimed to investigate the effects and mechanism of SOCE inhibition on liver injury induced by ethanol in BRL cells and Sprague-Dawley rats. Our data demonstrated that ethanol (0-400mM) dose-dependently increased hepatocyte injury and 100mM ethanol significantly upregulated the mRNA and protein expression of SOC for at least 72h in BRL cells. Blockade of SOCE by pharmacological inhibitors and sh-RNA knockdown of STIM1 and Orai1 attenuated intracellular Ca(2+) overload, restored the mitochondrial membrane potential (MMP), decreased cytochrome C release and inhibited ethanol-induced apoptosis. STIM1 and Orai1 expression was greater in ethanol-treated than control rats, and the SOCE inhibitor corosolic acid ameliorated the histopathological findings and alanine transaminase and aspartate transaminase activity as well as decreased cytochrome C release and inhibited alcohol-induced cell apoptosis. These findings suggest that SOCE blockade could alleviate alcohol-induced hepatotoxicity via inhibiting apoptosis. SOCE might be a useful therapeutic target in alcoholic liver diseases.
细胞外钙离子内流被认为在乙醇诱导的肝细胞凋亡和坏死中发挥作用。先前的研究表明，储存式钙离子内流（SOCE）参与了乙醇诱导的HepG2细胞肝损伤。然而，SOCE导致肝损伤的潜在机制仍不清楚。我们旨在研究SOCE抑制对BRL细胞和Sprague-Dawley大鼠乙醇诱导的肝损伤的影响及机制。我们的数据表明，乙醇（0 - 400mM）剂量依赖性地增加肝细胞损伤，100mM乙醇在BRL细胞中至少72小时显著上调SOC的mRNA和蛋白表达。药理学抑制剂阻断SOCE以及STIM1和Orai1的短发夹RNA敲低可减轻细胞内钙离子过载，恢复线粒体膜电位（MMP），减少细胞色素C释放并抑制乙醇诱导的凋亡。乙醇处理的大鼠中STIM1和Orai1表达高于对照大鼠，SOCE抑制剂科罗索酸改善了组织病理学表现以及丙氨酸转氨酶和天冬氨酸转氨酶活性，同时减少细胞色素C释放并抑制酒精诱导的细胞凋亡。这些发现表明，阻断SOCE可通过抑制凋亡减轻酒精诱导的肝毒性。SOCE可能是酒精性肝病的一个有用治疗靶点。
