硫代乙酰胺诱导慢性肝损伤小鼠肝组织YAP信号对胆管反应的影响及其分子机制  

Impact of YAP signaling on ductular reaction in mice with chronic liver injury and its molecular mechanism

作　　者：杨金连 张涛 莫小兰 YANG Jinlian;ZHANG Tao(Department of Pharmacy,Guangzhou Women and Children's Medical Center,Guangzhou Medical University,Guangzhou,Guangdong 510623,China)

机构地区：[1]广州医科大学附属妇女儿童医疗中心药学部,广东广州510623

出　　处：《医药前沿》2024年第20期1-5,共5页Journal of Frontiers of Medicine

基　　金：广东省基础与应用基础研究基金青年基金(2020A1515110314);广东省医院药学研究基金澳美基金(2023A27);广州医科大学附属妇女儿童医疗中心内部科研基金国自然青年培育基金-药学部专项(GWCMC2020-5-002)。

摘　　要：目的:探讨Yes相关蛋白(YAP)信号对硫代乙酰胺(TAA)诱导慢性肝损伤模型小鼠肝组织胆管反应(DR)的影响及其可能的分子机制。方法:选择野生型C57BL/6小鼠为实验对象,将TAA 300 mg/L喂养6周并每周予0.9%氯化钠溶液尾静脉注射小鼠作为TAA组,将TAA喂养6周并每周予YAP活性抑制剂维替泊芬(VP)尾静脉注射小鼠作为VP组,另设蒸馏水喂养的Control组。HE染色和天狼猩红染色观察小鼠肝组织形态学变化。采用Western blot检测肝组织YAP和肝细胞核因子4α(HNF4α)蛋白表达水平,采用qPCR检测YAP、HNF4α、卵圆细胞(OCs)标志分子A6、细胞增殖标志分子Ki-67、肝细胞标志分子清蛋白(ALB)和胆管上皮细胞(BECs)标志分子SOX9的mRNA表达水平;采用电镜观察肝组织中紧密连接的形成,采用免疫组织化学法检测肝组织细胞角蛋白19(CK19)表达水平。结果:TAA组肝组织DR较对照组明显,YAP蛋白和mRNA表达明显增加,HNF4α蛋白和mRNA显著降低。同时,A6和Ki-67的mRNA表达均明显增加,ALB mRNA表达明显降低,SOX9 mRNA表达明显增加。抑制YAP活性后,HNF4α蛋白和mRNA明显增加,A6 mRNA表达无明显改变,Ki-67 mRNA表达下降,但ALB mRNA表达明显增加,SOX9 mRNA表达明显降低。结论:TAA诱导小鼠慢性肝损伤模型中,YAP可能通过下调HNF4α的表达来抑制OCs分化为肝细胞,且促进OCs分化为BECs,进而诱导DR的发生而促进肝纤维化形成。Objective The aim of this experiment was to investigate the impact of Yes-associated protein(YAP)signaling on hepatic ductular reaction(DR)in mice with thioacetamide(TAA)-induced chronic liver injury and its possible molecular mechanism.Methods Wild-type C57BL/6 mice were selected as experimental subjects.Mice in TAA group were fed with TAA 300 mg/L for 6 weeks.The mice in VP group were fed with TAA 300 mg/L for 6 weeks,and injected with a YAP activity inhibitor Vitipofin(VP)via the tail vein twice a week.Mice in the control group were fed with distilled water.HE staining and Sirius scarlet staining were applied to observe the changes of liver histomorphology.The Western blot and RT-PCR were applied to detect YAP and hepatocyte nuclear factor 4 alpha(HNF4α)protein and gene levels in liver tissue.The gene levels of the mRNA expression of oval cells(OCs)marker A6,cell proliferation marker Ki-67,hepatocyte marker albumin(ALB)and bile duct epithelial cells(BECs)marker SOX9 were also detected.The formation of tight junctions was observed with electron microscopeused,the expression and distribution of DR marker CK19 was observed with immunohistochemical staining.Results Compared with the Control group,DR in the liver tissue of mice by feeding with a TAA diet for 6weeks increased significantly,the mRNA and the protein expression of YAP increased significantly,the mRNA and the protein expression of HNF4αdecreased significantly.Meanwhile,the mRNA expression of A6,Ki-67 and SOX9 increased significantly,the mRNA ALB decreased significantly.After inhibition of YAP activity,HNF4αprotein and mRNA increased significantly,the mRNA expression of A6 did not change significantly,and the mRNA expression of Ki-67 and SOX9 decreased,but ALB mRNA increased significantly.Conclusions In the chronic liver injury mouse model induced with a TAA diet,YAP may inhibit the differentiation of OCs into hepatocytes and promote the differentiation of OCs into BECs by down-regulating the expression of HNF4α,and then induce the occurrence of DR a

关 键 词：Yes相关蛋白 肝细胞核因子4Α 胆管反应 卵圆细胞 

分 类 号：R575.7[医药卫生—消化系统]