摘要
目的探讨哺乳动物ste20样激酶1(mammalian sterile20-like 1,MST1)基因的miRNAs对肝脏内脂质堆积的影响。方法利用高脂诱导非酒精性脂肪性肝病(NAFLD)模型小鼠与低脂对照组小鼠的肝脏及脂肪组织进行miRNAsequence分析,选取表达上调的miRNAs,通过数据库miRDB、starBase筛选出可能靶向MST1靶基因3’UTR区的miRNAs进行研究,利用Western blot及双荧光素酶报告基因实验检测miRNAs对MST1的靶向效应。并在NAFLD细胞模型内进行MST1过表达或干扰,进一步验证miRNAs对MST1的表达调控作用。结果 Western blot实验显示得到mmu miR 149-5p和mmu miR 499-5p抑制鼠MST1基因蛋白水平的表达,Luciferases实验证明mmu miR 149-5p和mmu miR 499-5p靶向MST1能够增加AML-12细胞内的脂质堆积。结论mmu miR 149-5p、mmu miR 499-5p能够调控小鼠肝脏MST1表达并影响肝脏内脂质堆积。
Objective To screen miRNAs that regulate mammalian ste20-like kinase 1(MST1)gene from liver and fat tissue of nonalcoholic fatty liver disease(NAFLD)model,and detect its effect on lipid accumulation in the liver.Methods miRNAs sequence analysis was performed on the liver and adipose tissue of NAFLD model mice induced by high-fat and low-fat control group.The miRNAs with up-regulated expression were selected and screened out by miRDB and starBase database.The targeting effect of miRNAs on MST1 was detected by Western blot and double luciferase reporter gene experiment.Furthermore,MST1 was overexpressed or interfered in NAFLD cell model to further verify the regulatory effect of miRNAs on MST1 expression.Results Western blot experiments showed that mmu miR 149-5p and mmu miR 499-5p inhibited the expression of murine MST1 gene protein,then Luciferases experiment proved that mmu miR 149-5p and mmu miR 499-5p could target MST1 to increase lipid accumulation in AML-12 cells.Conclusion Mmu miR 149-5p and mmu miR 499-5p can regulate MST1 expression in mouse liver and affect lipid accumulation in the liver.
作者
祁慧
李雨涵
梁涵子
李家瑞
李建宁
宋辉
杨怡
QI Hui;LI Yuhan;LIANG Hanzi;LI Jiarui;LI Jianning;SONG Hui;YANG Yi(Department of Biochemistry and Molecular Biology,School of Basic Medical Science,Ningxia Medical University,Yinchuan750004,China;Institute ofEndocrinology,Ningxia Medical University,Yinchuan 750004,China;Department of Cell Biology and Genetics,School of Basic Medical Science,Ningxia Medical University,Yinchuan750004,China)
出处
《宁夏医科大学学报》
2020年第7期649-658,共10页
Journal of Ningxia Medical University
基金
国家自然科学基金(81670798)。
