摘要
目的探讨哺乳动物不育系20样激酶1(MST1)在小鼠肝脏糖异生途径中的作用及调节机制。方法RT-qPCR检测MST1 mRNA在糖尿病模型db/db小鼠和对照db/m小鼠肝脏组织中的表达。通过腺病毒感染小鼠肝原代细胞,检测MST1过表达后肝细胞的糖输出能力。在HepG2细胞中,运用荧光素酶报告基因实验检测MST1对磷酸烯醇式丙酮酸羧化酶(PEPCK)编码基因的启动子活性的作用。利用干扰腺病毒在小鼠肝原代细胞中敲低叉头盒转录因子O1(FOXO1),检测MST1过表达对PEPCK mRNA水平的影响。结果MST1 mRNA在糖尿病db/db小鼠的肝脏组织中高表达(P<0.001)。在小鼠肝原代细胞过表达MST1促进了葡萄糖的输出(P<0.05)。在HepG2中,过表达MST1促进了PEPCK mRNA的表达(P<0.001)和启动子活性(P<0.01)。进而,当敲低FOXO1表达后,MST1诱导PEPCK mRNA表达的作用消失。结论MST1促进PEPCK表达参与调控小鼠肝脏糖异生依赖于FOXO1。
Objective To investigate the role of mammalian sterile line 20-like kinas 1(MST1)and its regulatory mechanism in hepatic gluconeogenesis of mouse.Methods The mRNA expression of MST1 in the liver of db/db mice and db/m mice was detected by RT-qPCR.MST1 was overexpressed with adenovirus to detect glucose output capacity in mouse primary hepatocytes.Luciferase reporter assay was adopted to detect the effect of MST1 on phosphoenolpyruvate carboxykinase(PEPCK)promoter activity in HepG2 cells.Forkhead transcription factor O1(FOXO1)was knocked down by interfering adenovirus in primary mouse liver cells to detect the effect of MST1 overexpression on the mRNA levels of PEPCK.Results The mRNA level of MST1 was significantly upregulated in the liver of db/db mice(P<0.001).MST1 overexpression enhanced glucose output in mouse primary hepatocytes(P<0.05).In HepG2 cells,MST1 overexpression increased the mRNA of PEPCK(P<0.001)and the activity of PEPCK promoter(P<0.01).Furthermore,MST1-induced increase in PEPCK mRNA was abolished when FOXO1was knocked down by interfering adenovirus.Conclusions MST1 promotes hepatic gluconeogenesis by regulating the mRNA expression of PEPCK dependent on FOXO1.
作者
解相宏
耿超
赵微
李春美
张伟虹
杨佳卉
刘晓军
XIE Xiang-hong;GENG Chao;ZHAO Wei;LI Chun-mei;ZHANG Wei-hong;YANG Jia-hui;LIU Xiao-jun(State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC,Beijing 100005;Department of Microbiology and Immunology, Basic Medicine College, Shanxi Medical University, Taiyuan 030001, China)
出处
《基础医学与临床》
CSCD
2020年第12期1656-1660,共5页
Basic and Clinical Medicine
基金
中国医学科学院医学与健康科技创新工程(CIFMS2017-I2M-1-008)。