Xin Xin, Cai Bei-Yu, Chen Cheng, Tian Hua-Jie, Wang Xin, Hu Yi-Yang, Feng Qin
Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Pudong New Area, Shanghai, 201203 China.
Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai, 201203 China.
Nutr Metab (Lond). 2020 May 26;17:40. doi: 10.1186/s12986-020-00462-y. eCollection 2020.
Even Non-alcoholic steatohepatitis (NASH) has been becoming the key role in process of liver fibrosis or cirrhosis, no any NASH involving liver fibrosis mice model which consistent with the mechanisms of fatty acid and glucose metabolism disorder was widely accepted. Here, we established a mouse model of nonalcoholic steatohepatitis (NASH) with liver fibrosis using a high-fat, high-carbohydrate diet (HFHC) and analyzed the potential pathogenesis using a transcriptome microarray.
Fifty mice were stratified by weight and randomly divided into the HFHC model and control (Con) groups. Ten mice were sacrificed at the beginning of the experiments, 10 mice of HFHC and Con group were euthanized at the end of 20 and 30 weeks. The following analyses were performed: biochemical analysis; histological assessment; evaluation of hepatic type I collagen (Col-I), α-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1) protein and mRNA expression levels; and transcriptomic gene chip analysis.
Compared with the Con group at each time point, the body weight and liver wet weight of the HFHC model group of mice were significantly higher. At 30th weeks, alanine aminotransferase (ALT), aspartate aminotransferase (AST), fasting blood glucose (FBG) and fasting insulin (FINS) levels or activities and the triglyceride (TG) and hydroxyproline (HYP) content in the HFHC model group were significantly elevated. Severe steatosis was present in the liver tissues contributed from the HFHC group of mice. Typically, substantial perisinusoidal fibrosis with a cage-like structure and bridging formations were observed in the mice liver in HFHC group. Col-I, α-SMA and TGF-β1 protein and mRNA expression levels in liver tissues of HFHC mice dramatically increased over time. Compared with the Con group, the HFHC group had 151 differentially expressed genes that were involved in 41 signaling pathways.
After keeping 30 weeks HFHC diet treatment, the mice exhibited substantial liver fibrosis, hepatic steatosis, ballooning degeneration and inflammation. Basing on the transcriptome microarray assays, the experimental NASH involving liver fibrosis potentially related to dramatically changed ECM-receptor interaction, Toll-like receptor signaling and other signaling pathways.
尽管非酒精性脂肪性肝炎（NASH）在肝纤维化或肝硬化进程中已成为关键因素，但尚无任何符合脂肪酸和葡萄糖代谢紊乱机制的NASH相关肝纤维化小鼠模型被广泛接受。在此，我们使用高脂高碳水化合物饮食（HFHC）建立了一种伴有肝纤维化的非酒精性脂肪性肝炎（NASH）小鼠模型，并通过转录组微阵列分析其潜在发病机制。
将50只小鼠按体重分层，随机分为HFHC模型组和对照组（Con）。实验开始时处死10只小鼠，在20周和30周结束时分别对HFHC组和Con组的10只小鼠实施安乐死。进行了以下分析：生化分析；组织学评估；评估肝I型胶原（Col-I）、α-平滑肌肌动蛋白（α-SMA）和转化生长因子-β1（TGF-β1）的蛋白质和mRNA表达水平；以及转录组基因芯片分析。
在每个时间点，与Con组相比，HFHC模型组小鼠的体重和肝脏湿重显著更高。在第30周时，HFHC模型组的丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、空腹血糖（FBG）和空腹胰岛素（FINS）水平或活性以及甘油三酯（TG）和羟脯氨酸（HYP）含量均显著升高。HFHC组小鼠的肝脏组织出现严重脂肪变性。典型的是，在HFHC组小鼠肝脏中观察到大量具有笼状结构和桥接形成的窦周纤维化。HFHC小鼠肝脏组织中Col-I、α-SMA和TGF-β1的蛋白质和mRNA表达水平随时间显著增加。与Con组相比，HFHC组有151个差异表达基因，涉及41条信号通路。
经过30周的HFHC饮食治疗后，小鼠出现大量肝纤维化、肝脂肪变性、气球样变性和炎症。基于转录组微阵列分析，实验性NASH相关肝纤维化可能与细胞外基质受体相互作用、Toll样受体信号传导和其他信号通路的显著变化有关。
