期刊论文

METTL3 regulates SPRING1 and promotes lipid accumulation in macrophages

中文

中国临床解剖学杂志

1001-165X

2021

39

06

686-691

10.13418/j.issn.1001-165x.2021.06.013

2020JJ4532:湖南省自然科学基金 18B264:湖南省教育厅优秀青年项目

本校为第一机构

目的 探讨METTL3调控SPRING1促进巨噬细胞脂质蓄积的作用及机制.方法 100 nig/mL PMA诱导THP-1细胞贴壁后,50 μg/mL Ac-LDL孵育THP-1细胞.Western blot测定METTL3和SPRING1蛋白;qRT-PCR测定SPRING1mRNA水平;细胞内总胆固醇、胆固醇酯以及游离胆固醇变化用高效液相色谱法检测;SRAMP和RMBase网站分析SPRING1 mRNA上的m6A修饰位点情况;质膜红色荧光标记探针Dil-Ac-LDL观察巨噬细胞脂滴摄取情况.结果 与对照组相比,Ac-LDL孵育后THP-1细胞METTL3和SPRING1蛋白表达上调,并且SPRING1 mRNA水平上调;过表达METTL3会使SPRING1蛋白表达上调,巨噬细胞对脂质摄取增加,细胞内Dil-Ac-LDL明显增多;反之,沉默MET...

Objective To investigate the effect and mechanism of METTL3 regulating SPRING1 on macrophage lipid accumulation.Methods After inducing the attachment of THP-1 cells with 100 ng/ml PMA,50 μg/ml Ac-LDL was added to incubate THP-1 cells.The protein levels of METTL3 and SPRING1 were detected by Western blot and the mRNA levels was detected by qRT-PCR.The total cholesterol(TC),cholesterol ester(CE)and free cholesterol(FC)were detected by high performance liquid chromatography(HPLC).SRAMP and RMBase websites were used to analyze the m6A modification...