miR-3077-5P对3T3-L1前脂肪细胞成脂分化能力的影响

Effects of miR-3077-5P on adipogenic differentiation of 3T3-L1 preadipocyte

目的观察miR-3077-5P对3T3-L1前脂肪细胞成脂分化能力的影响。方法通过细胞转染技术分别上调和下调3T3-L1前脂肪细胞中的miR-3077-5P表达,将其分为对照(control)组、模拟物(mimics)组及抑制物(inhibitor)组;将各组3T3-L1前脂肪细胞成脂诱导10 d,以逆转录聚合酶链式反应(Reverse transcription-polymerase chain reaction,RT-PCR)法检测其相关成脂基因PPAR-γ和LPL mRNA表达水平,用免疫蛋白印迹(Western Blot,WB)法检测其相关成脂基因PPAR-γ和FABP4蛋白水平;采用油红O染色观察脂滴形成情况并作定量分析。结果成脂诱导10 d,miR-3077-5P mimics组成脂基因PPAR-γ和LPL mRNA的表达水平及PPAR-γ和FABP4蛋白水平均升高,而miR-3077-5P inhibitor组则相反(P均<0. 05);成脂诱导10 d,miR-3077-5P mimics组脂滴形成最大,脂滴形成量存在升高现象,miR-3077-5P inhibitor组脂滴较小且形成量较少(P均<0. 05)。结论 miR-3077-5P对3T3-L1前脂肪细胞成脂分化过程具有促进作用。

Objective To investigate the effects of miR-3077-5P on 3T3-L1 adipogenic differentiation. Methods miR-3077-5P mimics,inhibitors and negative control were transfected into 3T3-L1 cells.The mRNA expressions of PPAR-γ and LPL were detected by real time RT-PCR and protein level of PPAR-γ and FABP4 were detected by Western Blot assay after adipogenic induction for 10 days.Oil-red staining was performed and quantified in 3T3-L1 cells. Results After adipogenic induction for 10 days,Overexpression of miR-3077-5P by the mimics,the mRNA expression of PPAR-γ and LPL and protein expression level of PPAR-γ and FABP4 were increased in miR-3077-5P mimics treatment group,whereas these genes expressions were decreased in miR-3077-5P inhibitors treatment group.Moreover,Oil-red staining showed that more formation of lipid droplets was shown in miR-3077-5P mimics treatment group than that in miR-3077-5P inhibitors treatment group. Conclusion miR-3077-5P could promote the adipogenic differentiation of 3T3-L1 cells.