中国医科大学学报2024，Vol.53Issue(5)：392-400,9.DOI:10.12007/j.issn.0258-4646.2024.05.002
YAP调控Notch信号通路影响小鼠NASH肝纤维化的机制及慈菇消脂方的干预作用
Mechanism of Notch signaling pathway regulated by YAP in NASH liver fibrosis mice and the intervention effect of Cigu Xiaozhi prescription
摘要
Abstract
Objective To investigate the regulatory mechanism of Notch signaling pathway by YAP in non-alcoholic steatohep-atitis(NASH)liver fibrosis,and assess the intervention effect of Cigu Xiaozhi prescription in detoxification and phlegm treatment.Methods C57BL/6J mice were randomly divided into different groups,including a normal group,NASH liver fibrosis model group,verteporfin(VP)intervention group,VP+Chinese medicine(Cigu Xiaozhi prescription)low-dose group,VP+Chinese medicine high-dose group,and dimethyl sulfoxide control group.The methionine/choline-deficient diet combined with low-dose CCl4 was used to construct the NASH liver fibrosis model.The degree of liver fibrosis was evaluated using hematoxylin and eosin(HE)and Masson staining.Four protein factors associated with liver fibrosis were detected using enzyme-linked immunosorbent assay,and hydroxyproline levels in the mouse liver was determined using the alkaline water method.The localization of α-SMA,ColⅠ,YAP,and Notch1 proteins in the liver was determined using immunohistochemistry.Additionally,the mRNA and protein expression levels of the Notch signaling pathway molecules,namely Notch1/2,J agged1,and DLL4,were assessed using real-time quantitative PCR and Western blotting analyses,respectively.Results The HE and Masson staining results revealed that the liver cells of NASH liver fibrosis mice were swollen and the cytoplasm was transparent.Additionally,evident fibrosis was observed in the hepatic lobule,portal area,and sinus;it was accompanied by heightened levels of inflam-matory cell infiltration,a large number of fat droplets,and instances of local hepatocyte necrosis,dissolution,and cirrhosis.The four factors associated with liver fibrosis showed a substantial increase(P<0.01).α-SMA,ColⅠ,YAP,and Notch1 were localized in the cytoplasm of hepatocytes.YAP,Notch1/2,and Jagged1 were highly expressed in the liver(P<0.01)but were downregulated after intervention with VP and VP+high and low doses of Cigu Xiaozhi prescription(P<0.05).Meanwhile,DLL4 factor was upregulated in the VP+high-dose of Cigu Xiaozhi prescription group(P<0.05).Conclusion YAP may inhibit activation of the Notch pathway by downregulating Notch1/2 and Jagged1 and upregulating DLL4,thereby interfering with the occurrence of liver fibrosis in NASH.Treatment with Cigu Xiaozhi pre-scription may inhibit Notch signaling pathway activation by downregulating YAP,Notch1/2,and Jagged1 and upregulating DLL4 through its multi-components and multi-targets properties,ultimately slow the progression of liver fibrosis in NASH.关键词
非酒精性脂肪性肝炎肝纤维化/Yes相关蛋白/Notch信号通路/慈菇消脂方Key words
non-alcoholic steatohepatitis liver fibrosis/Yes-associated protein/Notch signaling pathway/Cigu Xiaozhi prescription分类
医药卫生引用本文复制引用
王莉,王兴胜,赵秀萍,王爱娣,屈红梅,马燕花..YAP调控Notch信号通路影响小鼠NASH肝纤维化的机制及慈菇消脂方的干预作用[J].中国医科大学学报,2024,53(5):392-400,9.基金项目
国家自然科学基金地区基金(81860821) （81860821）
甘肃省科技计划(23JDKD0001) （23JDKD0001）
甘肃中医药大学第一临床医学院2021年度研究生创新基金(LCCX2021001) （LCCX2021001）
