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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 micro RNA-152 (mi R-152) on the cholesterol accumulation in RAW264.7 macrophages stimulated by ox-LDL and the determination of its target genes. Methods Mouse RAW264.7 macrophages were divided into two groups, mi R-152 group and negative control group. After stimulated by ox-LDL for 48 h, the content of total cholesterol (TC), cholesterolester (CE) and the ratio of TC / CE in each group were determined by enzyme-linked immunosorbent assay. Bioinformatics methods, dual luciferase reporter assay, real-time fluorescence quantitative PCR and Western blot confirmed the downstream targets of mi R-152. Results Compared with the control group, the levels of TC and CE in mi R-152 macrophages increased and the ratio of CE / TC increased significantly. Moreover, the mRNA and protein expression levels of ABCA1 in mi R-152 group were significantly higher than those in control group reduce. Conclusions mi R-152 can promote ox-LDL-induced cholesterol accumulation in RAW264.7 macrophages, and its mechanism may be related to the regulation of ABCA1.