CAJ | 학술논문

目的研究在3T3-L1脂肪细胞中G蛋白耦联受体120(GPR120)与胰岛素受体底物-1(IRS-1)的关系.方法利用经典“鸡尾酒法”诱导3T3-L1前脂肪细胞分化,以未诱导组作为阴性对照,通过油红O染色法检测细胞内脂滴的形成情况,从而确定前脂肪细胞分化为脂肪细胞,再利用实时PCR方法检测GPR120 mRNA的表达水平.采用siRNA技术下调3T3-L1前脂肪细胞中GPR120的表达,以无关干扰组为阴性对照,干扰24 h后诱导细胞分化,诱导后于3T3-L1脂肪细胞培养液中加入软脂酸孵育24 h,分别用实时PCR和Western印迹方法检测3T3-L1脂肪细胞中IRS-1的表达水平.结果诱导分化的3T3-L1脂肪细胞中,GPR120 mRNA表达量较未诱导组明显升高(P＜0.05);干扰GPR120表达后3T3-L1脂肪细胞中脂滴体积和数量明显减小.另外,GPR120表达的量降低后,IRS-1 mRNA和蛋白表达水平均显著降低(P＜0.05).结论 GPR120参与胰岛素信号通路中IRS-1的表达调控.
목적 연구재3T3-L1지방세포중G단백우련수체120(GPR120)여이도소수체저물-1(IRS-1)적관계.방법 이용경전“계미주법”유도3T3-L1전지방세포분화,이미유도조작위음성대조,통과유홍O염색법검측세포내지적적형성정황,종이학정전지방세포분화위지방세포,재이용실시PCR방법검측GPR120 mRNA적표체수평.채용siRNA기술하조3T3-L1전지방세포중GPR120적표체,이무관간우조위음성대조,간우24 h후유도세포분화,유도후우3T3-L1지방세포배양액중가입연지산부육24 h,분별용실시PCR화Western인적방법검측3T3-L1지방세포중IRS-1적표체수평.결과 유도분화적3T3-L1지방세포중,GPR120 mRNA표체량교미유도조명현승고(P＜0.05);간우GPR120표체후3T3-L1지방세포중지적체적화수량명현감소.령외,GPR120표체적량강저후,IRS-1 mRNA화단백표체수평균현저강저(P＜0.05).결론 GPR120삼여이도소신호통로중IRS-1적표체조공.
Objective To investigate the correlation between G protein-coupled receptor 120 (GPR120) and insulin receptor substrate-l(IRS-1) in 3T3-L1 cells.Methods A classic "cocktail" method was applied for induction of preadipocyte differentiation,and cells without induction were selected as a negative control.To determine the differentiation of preadipocyte to adipocyte,oil red O method was used to test the content of lipid droplet in cells.Real-time PCR was used to assess the expression of GRP120 mRNA.A specific siRNA was used to intervene the expression of GRP120 in 3T3-L1 preadipocyte,and no interfering group was used as a negative control.After interfering for 24 hours,3T3-L1 preadipocyte was incubated with palmitic acid for another 24 hours to induce the differentiation.Real-time PCR and Western blot were used to detect the expression of IRS-1 mRNA and protein.Results The mRNA level of GPR120 was up-regulated in differentiated 3T3-L1 preadipocyte compared with control group(P ＜ 0.05).The volume and quantity of lipid droplet in 3T3-L1 adipocyte were significantly decreased after interfering the expression of GPR120 expression.Furthermore,IRS-1 mRNA and protein levels were significantly reduced (P ＜ 0.05).Conclusion GPR120 plays a regulatory role in the expression of IRS-1 in insulin signaling pathway.