Xie Fang, Jia Lin, Lin Minghua, Shi Ying, Yin Jiming, Liu Yin, Chen Dexi, Meng Qinghua
Beijing You An Hospital, Affiliated Hospital of Capital Medical University, Beijing, China; Beijing Institute of Hepatology, Beijing, China.
J Cell Mol Med. 2015 Jan;19(1):155-64. doi: 10.1111/jcmm.12364. Epub 2014 Sep 25.
ASPP2 is a pro-apoptotic member of the p53 binding protein family. ASPP2 has been shown to inhibit autophagy, which maintains energy balance in nutritional deprivation. We attempted to identify the role of ASPP2 in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). In a NAFLD cell model, control treated and untreated HepG2 cells were pre-incubated with GFP-adenovirus (GFP-ad) for 12 hrs and then treated with oleic acid (OA) for 24 hrs. In the experimental groups, the HepG2 cells were pre-treated with ASPP2-adenovirus (ASPP2-ad) or ASPP2-siRNA for 12 hrs and then treated with OA for 24 hrs. BALB/c mice fed a methionine- and choline-deficient (MCD) diet were used to generate a mouse model of NAFLD. The mice with fatty livers in the control group were pre-treated with injections of GFP-ad for 10 days. In the experimental group, the mice that had been pre-treated with ASPP2-ad were fed an MCD diet for 10 days. ASPP2-ad or GFP-ad was administered once every 5 days. Liver tissue from fatty liver patients and healthy controls were used to analyse the role of ASPP2. Autophagy, apoptosis markers and lipid metabolism mediators, were assessed with confocal fluorescence microscopy, immunohistochemistry, western blot and biochemical assays. ASPP2 overexpression decreased the triglyceride content and inhibited autophagy and apoptosis in the HepG2 cells. ASPP2-ad administration suppressed the MCD diet-induced autophagy, steatosis and apoptosis and decreased the previously elevated alanine aminotransferase levels. In conclusion, ASPP2 may participate in the lipid metabolism of non-alcoholic steatohepatitis and attenuate liver failure.
ASPP2是p53结合蛋白家族的促凋亡成员。已证明ASPP2可抑制自噬，而自噬在营养缺乏时维持能量平衡。我们试图确定ASPP2在非酒精性脂肪性肝病（NAFLD）发病机制中的作用。在NAFLD细胞模型中，将对照处理和未处理的HepG2细胞与绿色荧光蛋白腺病毒（GFP - ad）预孵育12小时，然后用油酸（OA）处理24小时。在实验组中，将HepG2细胞用ASPP2腺病毒（ASPP2 - ad）或ASPP2小干扰RNA（siRNA）预处理12小时，然后用OA处理24小时。喂食蛋氨酸和胆碱缺乏（MCD）饮食的BALB / c小鼠用于建立NAFLD小鼠模型。对照组有脂肪肝的小鼠用GFP - ad注射预处理10天。在实验组中，用ASPP2 - ad预处理的小鼠喂食MCD饮食10天。每5天给予一次ASPP2 - ad或GFP - ad。使用脂肪肝患者和健康对照的肝组织来分析ASPP2的作用。通过共聚焦荧光显微镜、免疫组织化学、蛋白质免疫印迹和生化分析评估自噬、凋亡标志物和脂质代谢介质。ASPP2的过表达降低了HepG2细胞中的甘油三酯含量，并抑制了自噬和凋亡。给予ASPP2 - ad可抑制MCD饮食诱导的自噬、脂肪变性和凋亡，并降低先前升高的丙氨酸转氨酶水平。总之，ASPP2可能参与非酒精性脂肪性肝炎的脂质代谢并减轻肝衰竭。
