期刊论文

Sortilin alters lipid efflux from lipid-laden THP-1 macrophage by regulating ABCA1 protein

中文

中国临床解剖学杂志

1001-165X

2020

38

5

568-573

10.13418/j.issn.1001-165x.2020.05.015

国家自然科学基金(81770460) 加拿大萨斯喀彻温省卫生研究基金会博士后基金(SHRF4144) 衡阳市科学技术发展计划项目(2017KJ268、2017KJ182) 南华大学博士启动基金(2014XQD37) 大学生研究性学习和创新性实验计划项目(201810555015) 湖南省自然科学基金(2020JJ4532)。

本校为第一机构

目的探讨Sortilin对荷脂THP-1巨噬细胞ABCA1蛋白表达及胞内脂质流出的影响。方法采用Western blot和qRT-PCR检测细胞内ABCA1蛋白和mRNA表达水平;Co-IP实验检测Sortilin和ABCA1的结合情况;高效液相色谱法检测细胞内总胆固醇、游离胆固醇、胆固醇酯含量变化;油红O染色观察细胞内脂滴情况。结果 Sortilin下调细胞内ABCA1蛋白水平,但对其mRNA水平无显著影响;Co-IP结果表明Sortilin能与ABCA1结合;与对照组相比,Sortilin过表达后荷脂巨噬细胞内胆固醇流出减少,胞内脂质含量增加且脂滴肥大,数量明显增多,Sortilin沉默后荷脂巨噬细胞胆固醇流出增加,胞内脂质含量减少,脂滴瘦小,数量减少;溶酶体抑制剂氯喹...

Objective To investigate the effect of Sortilin on protein level of macrophage ABCA1 and intracellular lipid efflux. Methods The protein and mRNA levels of lipid-laden macrophage ABCA1 were detected by western blot(WB) and quantitative real-time polymerase chain reaction(qRT-PCR)respectively;the combination of Sortilin with ABCA1 was detected by co-immunoprecipitation(Co-IP);macrophage cholesterol efflux was measured by liquid scintillation counting. Total cholesterol(TC), free cholesterol(FC) and cholesterol ester(CE) were determined by high performance liquid chromatography(HPLC). Intracellu...