miR-23b-3p对3T3-L1前脂肪细胞增殖和成脂分化的影响

Role of miR-23b-3p in 3T3-L1 preadipocyte proliferation and differentiation

为探讨miR-23b-3p对3T3-L1前脂肪细胞增殖和成脂分化的影响,通过化学修饰的miR-23b-3p模拟物成功转入3T3-L1前脂肪细胞后,利用CCK-8法和流式细胞术检测过表达miR-23b-3p对3T3-L1前脂肪细胞增殖的影响;并通过实时荧光定量PCR和Western blot检测细胞周期基因CyclinD1和CDK4以及成脂标志基因PPARγ和FABP4的表达情况;油红O染色测定甘油三酯积累情况。结果显示,过表达miR-23b-3p抑制3T3-L1前脂肪细胞增殖,导致细胞G1期阻滞,CyclinD1、CDK4在mRNA和蛋白水平极显著下调;在3T3-L1前脂肪细胞成脂分化过程中,miR-23b-3p的表达水平上调;过表达miR-23b-3p促进3T3-L1前脂肪细胞甘油三酯积累,PPARγ、FABP4在mRNA和蛋白水平显著或极显著上调。结果表明,miR-23b-3p能够抑制3T3-L1前脂肪细胞增殖并促进其分化为成熟脂肪细胞。

To explore the influence of mir-23b-3p on 3T3-L1 proliferation and adipocyte differentia tion. Following the transfection of miR-23b-3p mimic into 3T3-L1 cells,cck-8 and flow cytometry were used to measure the effect of miR-23b-3p on 3T3-L1 preadipocyte proliferation; the expres- sion of cell cycle genes CyclinD1,CDK4 and adipogenic marker genes PPART,FABP4 were detec- ted by real-time qPCR and Western blot;oil red O staining was used to examine the content of tri- glyceride. Overexpression of miR-23b-3p inhibited 3T3-L1 preadipocyte proliferation and blocked the G1 phase;the expression of CyclinD1 and CDK4 was significantly reduced at mRNA and pro- tein levels. The miR-23b-3p expression was up-regulated during the differentiation of 3T3-L1 prea- dipocytes; overexpression of miR-23b-3p promoted adipogenesis by increasing the mRNA and pro- tein expression of adipogenic marker genes, including peroxisome proliferators-activated receptor 7,fatty acid-binding protein 4, as well as triglyceride accumulation. In conclusion, the miR-23b-3p can inhibit 3T3-L1 preadipocyte proliferation and promote adipogenic differentiation.