Liu Xiaoying, Henkel Anne S, LeCuyer Brian E, Schipma Matthew J, Anderson Kristy A, Green Richard M
Division of Gastroenterology and Hepatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois; and
Division of Gastroenterology and Hepatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois; and.
Am J Physiol Gastrointest Liver Physiol. 2015 Dec 15;309(12):G965-74. doi: 10.1152/ajpgi.00132.2015. Epub 2015 Oct 15.
Fatty liver is associated with endoplasmic reticulum stress and activation of the hepatic unfolded protein response (UPR). Reduced hepatic expression of the UPR regulator X-box binding protein 1 spliced (XBP1s) is associated with human nonalcoholic steatohepatitis (NASH), and feeding mice a high-fat diet with fructose/sucrose causes progressive, fibrosing steatohepatitis. This study examines the role of XBP1 in nonalcoholic fatty liver injury and fatty acid-induced cell injury. Hepatocyte-specific Xbp1-deficient (Xbp1(-/-)) mice were fed a high-fat/sugar (HFS) diet for up to 16 wk. HFS-fed Xbp1(-/-) mice exhibited higher serum alanine aminotransferase levels compared with Xbp1(fl/fl) controls. RNA sequencing and Gene Ontogeny pathway analysis of hepatic mRNA revealed that apoptotic process, inflammatory response, and extracellular matrix structural constituent pathways had enhanced activation in HFS-fed Xbp1(-/-) mice. Liver histology demonstrated enhanced injury and fibrosis but less steatosis in the HFS-fed Xbp1(-/-) mice. Hepatic Col1a1 and Tgfβ1 gene expression, as well as Chop and phosphorylated JNK (p-JNK), were increased in Xbp1(-/-) compared with Xbp1(fl/fl) mice after HFS feeding. In vitro, stable XBP1-knockdown Huh7 cells (Huh7-KD) and scramble control cells (Huh7-SCR) were generated and treated with palmitic acid (PA) for 24 h. PA-treated Huh7-KD cells had increased cytotoxicity measured by lactate dehydrogenase release, apoptotic nuclei, and caspase3/7 activity assays compared with Huh7-SCR cells. CHOP and p-JNK expression was also increased in Huh7-KD cells following PA treatment. In conclusion, loss of XBP1 enhances injury in both in vivo and in vitro models of fatty liver injury. We speculate that hepatic XBP1 plays an important protective role in pathogenesis of NASH.
脂肪肝与内质网应激及肝脏未折叠蛋白反应(UPR)的激活有关。UPR调节因子X盒结合蛋白1剪接体(XBP1s)的肝脏表达降低与人非酒精性脂肪性肝炎(NASH)相关,给小鼠喂食含果糖/蔗糖的高脂饮食会导致进行性纤维化脂肪性肝炎。本研究探讨XBP1在非酒精性脂肪肝损伤和脂肪酸诱导的细胞损伤中的作用。给肝细胞特异性Xbp1基因缺陷(Xbp1(-/-))小鼠喂食高脂/高糖(HFS)饮食长达16周。与Xbp1(fl/fl)对照小鼠相比,喂食HFS的Xbp1(-/-)小鼠血清丙氨酸转氨酶水平更高。对肝脏mRNA进行RNA测序和基因本体途径分析发现,在喂食HFS的Xbp1(-/-)小鼠中,凋亡过程、炎症反应和细胞外基质结构成分途径的激活增强。肝脏组织学显示,喂食HFS的Xbp1(-/-)小鼠的损伤和纤维化增强,但脂肪变性较少。与喂食HFS后的Xbp1(fl/fl)小鼠相比,Xbp1(-/-)小鼠肝脏中Col1a1和Tgfβ1基因表达以及Chop和磷酸化JNK(p-JNK)增加。在体外,构建了稳定敲低XBP1的Huh7细胞(Huh7-KD)和对照乱序细胞(Huh7-SCR),并用棕榈酸(PA)处理24小时。与Huh7-SCR细胞相比,PA处理的Huh7-KD细胞通过乳酸脱氢酶释放、凋亡细胞核和caspase3/7活性测定显示细胞毒性增加。PA处理后,Huh7-KD细胞中CHOP和p-JNK表达也增加。总之,XBP1缺失在体内和体外脂肪肝损伤模型中均增强了损伤。我们推测肝脏XBP1在NASH发病机制中起重要保护作用。