microRNA-152对影RAW264.7巨噬细胞胆固醇蓄积的响及其靶基因验证

Effect of microRNA-152 on cholesterol accumulation in RAW264.7 and identification of its functional target genes

目的探讨micro RNA-152(mi R-152)对ox-LDL刺激下的RAW264.7巨噬细胞胆固醇蓄积的影响及其靶基因的确定。方法小鼠RAW264.7巨噬细胞分为两组,mi R-152组和阴性对照组。ox-LDL刺激两组细胞48 h后,酶荧光学法测定各组细胞内总胆固醇(total cholesterol,TC)、胆固醇酯(cholesterolester,CE)的含量和TC/CE比例。生物信息学方法、双荧光素酶报告及实时荧光定量PCR和Western blot证实mi R-152的下游靶基因。结果与对照组相比,mi R-152组巨噬细胞内TC和CE含量增加,CE/TC的比值明显升高;并且,mi R-152组ABCAl的m RNA和蛋白表达水平较对照组明显降低。结论 mi R-152能够促进ox-LDL诱导的RAW264.7巨噬细胞胆固醇蓄积,其机制可能与靶向调控ABCA1有关。

Objective To investigate the effect of miR-152 on ox-LDL induced cholesterol accumulation in RAW264.7 macrophages and to verify its target genes. Methods RAW264.7 macrophages were divided into two groups： miR-152 group and negative control group. Total cholesterol （TC）, cholesterol ester （CE） eoncentra-tion and the ratio of CE/TC were observed in the two group by ox-LDL for 48 h. Furthermore, bioinformatics meth- od, dual luciferase reporter assay, real-time quantitative PCR and Western blot were applied to detect the target gene of miR- 152. Results As compared with the control group, the contents of TC, CE and the ratio of CEfFC were increased in the miR-152 group. The mRNA and protein expressions of ABCA1 were significantly lower in miR -152 group. Conclusions MiR-152 could inhibit macrophage foam cell formation through decreasing the expres- sion of ABCA1.