Mfn2基因对RAW264.7巨噬细胞源性泡沫细胞形成及ABCA1表达的影响

The effect of Mfn2 on expression of ABCA1 and RAW264.7 macrophage derived foam cell formation

目的:观察Mfn2基因对ox-LDL刺激下RAW264.7巨噬细胞源性泡沫细胞形成的影响及其机制。方法:通过细胞内胆固醇浓度检测,分析过表达或者干扰Mfn2基因表达对RAW264.7细胞在ox-LDL刺激下泡沫细胞内胆固醇含量和胆固醇酯/总胆固醇比例的影响;通过实时定量PCR和Western blot检测Mfn2基因对细胞胆固醇外排蛋白ABCA1mRNA和蛋白水平的影响。结果:RAW264.7细胞过表达Mfn2基因,分别降低细胞内16.3%总胆固醇含量和19.8%胆固醇酯/总胆固醇比例;而干扰RAW264.7细胞Mfn2基因表达,分别增加细胞内45.9%总胆固醇含量和26.7%的胆固醇酯/总胆固醇比例。40pfu和60pfu adv-Mfn2感染均促进RAW264.7细胞ABCA1的mRNA表达(分别增加26.5%和60.5%),而仅60pfu adv-Mfn2感染促进RAW264.7细胞ABCA1的蛋白表达(增加40.6%)。结论:Mfn2可能通过促进ABCA1的表达,减少细胞内胆固醇的沉积。

Objective：To investigate the effect of Mfn2 gene on ox-LDL induced RAW264.7 macrophage de- rived foam cell formation. Method：Intracellular cholesterol concentration and intracellular cholesterol ester / total cholesterol ratio was detected by the cellular cholesterol assay kit, confirm the effect of Mfn2 on intraeellular lipid deposition in RAW264.7 cells under oxLDL stimulation. In further, the effect of Mfn2 gene on mRNA and pro- tein expression of cholesterol transporter ABCA1 was detected by real-time quantitative PCR and western blot sep- arately. Result：Overexpression or silence of Mfn2 gene expression reduces or increases cholesterol accumulation and cholesterol ester / free cholesterol ratio in RAW264.7 derived foam cells. ABCA1 mRNA was increased by 40 pfu or 60 pfu Adv-Mfn2 infection, but the protein of ABCA1 was only increased by 60 pfu Adv-Mfn2 infection. Conclusion ： Overexpression of Mfn2 reduces intracellular cholesterol deposition by promoting the expression of AB- CA1.