基于RNA-seq转录组学分析一贯煎对酒精性脂肪性肝病小鼠TGFBR2受体表达的影响  

Effects of Yiguan Decoction on TGFBR2 Receptor of Mice with Alcoholic Fatty Liver Disease Based on RNA-seq Transcriptomic Analysis

作　　者：邱丰俊 李杜 朱一唯 闫晓风[1] 叶䁎杰 王晓玲[1] 胡旭东[1] QIU Fengjun;LI Du;ZHU Yiwei;YAN Xiaofeng;YE Tingjie;WANG Xiaoling;HU Xudong(School of Basic Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《中国中医药信息杂志》2021年第5期42-49,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金(81703832);上海中医药大学预算内科研项目(2020LK002)。

摘　　要：目的通过RNA-seq转录组学测序探讨一贯煎防治小鼠酒精性脂肪性肝病(AFLD)的潜在信号通路。方法制备小鼠AFLD模型,同时予一贯煎药液灌胃。HE染色观察小鼠肝脏损伤情况,检测小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、三酰甘油(TG)含量,油红O染色观察肝脏脂肪生成情况,RNA-seq转录组测序技术检测小鼠肝脏差异表达基因,并进行KEGG通路富集分析,同时以RT-qPCR进行验证。体外实验以酒精刺激AML12细胞诱导AFLD细胞模型,通过TGFBR2和SLC2A4抗体拮抗相应的肝细胞膜受体,同时给予一贯煎溶液干预,Nile Red荧光染色观察细胞内脂质合成变化。结果HE染色显示,一贯煎可减轻AFLD小鼠肝损伤;油红O染色显示,一贯煎可降低肝组织内脂质沉积;与对照组比较,模型组小鼠血清ALT、TG含量明显增加(P<0.0001),与模型组比较,一贯煎组小鼠血清ALT、TG含量减少(P<0.0001),模型组、一贯煎组较对照组血清AST含量增加,差异无统计学意义(P>0.05);KEGG通路富集结果表明,FOXO信号通路中细胞膜受体蛋白TGFBR2、SLC2A4可能是一贯煎防治小鼠AFLD的关键信号通路蛋白。体外细胞实验表明,一贯煎和TGFBR2抗体单用或联用均能显著降低酒精诱导的AML12细胞脂肪变性,SLC2A4抗体对酒精诱导AML12细胞脂肪变性无明显影响。结论一贯煎可能通过影响肝细胞膜受体蛋白TGFBR2的表达而起到防治AFLD的药理效应。Objective To explore the potential signal pathway of Yiguan Decoction in the prevention and treatment of mice with alcoholic fatty liver disease(AFLD)through RNA-seq transcriptomic sequencing.Methods AFLD mice model was established.The model received gavage by Yiguan Decoction.HE staining was used to observe the liver damage in mice,and the kit was used to detect serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and triacylglycerol(TG)levels in mice.Oil red O staining was used to observe liver lipogenesis;RNA-seq transcriptome sequencing technology was used to detect differentially expressed genes in mouse liver;KEGG pathway enrichment analysis was performed;RT-qPCR was used for verification.In vitro experiments stimulated AML12 cells with alcohol to induce AFLD cell model,antagonized the corresponding liver cell membrane receptors through TGFBR2 and SLC2A4 antibodies,and Yiguan Decoction was given for intervention at the same time.The changes in lipid synthesis in cells were observed by Nile Red fluorescent staining.Results The results of HE staining showed that Yiguan Decoction could significantly reduce liver injury in AFLD mice.Oil red O staining showed that Yiguan Decoction could reduce lipid deposition in liver tissue;Compared with the control group,the serum ALT and TG contents of mice in the model group significantly increased(P<0.0001).Compared with the model group,the serum ALT and TG contents of mice in the Yiguan Decoction group decreased(P<0.0001).The serum AST content of the model group and Yiguan Decoction group was higher than that of the control group,without statistical significance(P>0.05).The enrichment results of the KEGG pathway indicated that the membrane receptor proteins TGFBR2 and SLC2A4 in the FOXO signaling pathway may be the key signaling pathway proteins for Yiguan Decoction to prevent and treat AFLD in mice.In vitro cell experiments showed that Yiguan Decoction and TGFBR2 antibody alone or in combination could significantly reduce alcohol-induced fatty degeneration

关 键 词：一贯煎 酒精性脂肪性肝病 RNA-SEQ TGFBR2 脂肪变性 小鼠 AML12细胞 

分 类 号：R285.5[医药卫生—中药学]