胆酸通过上调miR-23b-3p抑制载脂蛋白A表达  

Bile Acid Inhibits Apolipoprotein A Expression in Hep G2 Cells by Up-regulation of miR-23b-3p

作　　者：田永凤[1,2] 莫学靓[2] 雷建军[2] 刘亚密 白洁[3] 王佐[2] 

机构地区：[1]山东省临沂市妇女儿童医院病理科,山东省临沂市276000 [2]南华大学心血管疾病研究所动脉硬化学湖南省重点实验室 [3]南华大学附属第一医院功能科,湖南省衡阳市421001

出　　处：《中国动脉硬化杂志》2016年第8期769-773,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81070221);湖南省卫生厅课题(B2013-034)

摘　　要：目的分析胆酸降载脂蛋白A(Apo A)效应与miR-23b-3p的关系,研究胆酸降Apo A作用新机制。方法首先用生物信息学在线工具对miR-23b-3p与调控LPA基因的转录因子肝细胞核因子4γ(HNF4γ)进行靶基因分析,使用荧光素酶报告系统对miR-23b-3p与调控LPA基因的转录因子HNF4γ进行靶基因验证实验,Western blot检测Apo A表达水平、p38MAPK(MAPK:丝裂原活化蛋白激酶)及p-p38MAPK,实时定量PCR检测miR-23b-3p表达水平。结果生物信息学分析表明HNF4γ可作为miR-23b-3p的靶基因,荧光素酶报告系统转染miR-23b-3p处理组细胞裂解后荧光强度显著低于对照组,验证了HNF4γ可作为miR-23b-3p的靶基因。胆酸呈剂量和时间依赖性抑制Hep G2细胞Apo A的表达,以32 mg/L和24 h的作用最显著。胆酸抑制Apo A表达与活化MAPK和上调miR-23b-3p有关。结论胆酸呈剂量和时间依赖性地下调Hep G2细胞Apo A表达水平;胆酸降Apo A与上调miR-23b-3p有关。Aim To analyze the relationship between the effect of bile acid reducing the level of apolipoprotein A（ Apo A） and the expression of miR-23b-3p,and to find new mechanisms of bile acid reducing the level of Apo A.Methods The target genes of miR-23b-3p and transcription factor hepatocyte nuclear factor 4γ（ HNF4γ） regulating the LPA gene were analyzed by bioinformatics online tools Targetscan 7. 0. Luciferase reporter assay was used to test target gene relationship for miR-23b-3p. Protein expression of Apo A,mitogen-activated protein kinase（ p38MAPK） and pp38 MAPK were detected by Western blot in Hep G2 cells,and miR-23b-3p expression was measured by real-time quantitative polymerase chain reaction. Results Bioinformatics analysis indicated that HNF4γ could act as a target gene of miR23b-3p. The fluorescent intensity of miR-23b-3p transfection cells was significantly lower than that of control group by using luciferase report assay,and it meant that HNF4γ could act as a target gene of miR23b-3p. Bile acid restrained Apo A expression in Hep G2 cells in a dose- and time-dependent manner,and 32 mg / L and 24 h were the best action dose and time. Bile acid restraining the Apo A expression was related to the activation of MAPK and the up-regulation of miR-23b-3p. Moreover,it could regulate the expressions of miR-23b-3p,farnesyl X receptor and MAPK. Conclusion Bile acid can significantly down-regulate Apo A expression in Hep G2 cells by a dose-and time-dependent manner,its mechanism is related to up-regulating the expression of miR-23b-3p.

关 键 词：胆酸 载脂蛋白A HEPG2细胞 miR-23b-3p 

分 类 号：R363[医药卫生—病理学]