Ⅲ型纤连蛋白组件包含蛋白5通过抑制ERK1/2磷酸化抑制C3H10T1/2细胞成脂分化  

FNDC5 Regulates the Adipogenic Differentiation of C3H10T1/2 Cells by Inhibiting the Phosphorylation of ERK1/2

作　　者：黑伟 何志强 张燕伟 张万锋 蔡春波 杨阳 高鹏飞[1] 郭晓红[1] 曹果清[1] 李步高[1] HEI Wei;HE Zhi-Qiang;ZHANG Yan-Wei;ZHANG Wan-Feng;CAI Chun-Bo;YANG Yang;GAO Peng-Fei;GUO Xiao-Hong;CAO Guo-Qing;LI Bu-Gao(College of Animal Sciences,Shanxi AgriculturalUniversity,Taigu 030801,Shanxi,China)

机构地区：[1]山西农业大学动物科学学院动物遗传育种与繁殖系实验室,山西太谷030801

出　　处：《中国生物化学与分子生物学报》2021年第5期644-652,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.31872336);三晋学者支持计划专项经费(2016,2017);山西省应用基础研究项目(No.201901D211376,201901D211369)。

摘　　要：旨在探究Ⅲ型纤连蛋白组件包含蛋白5(typeⅢdomain-containing protein5,FNDC5)对C3H10T1/2细胞成脂分化的调控作用。利用qRT-PCR和Western印迹检测FNDC5在C3H10T1/2细胞成脂分化过程中的时序性表达规律;构建慢病毒包被的过表达/干扰FNDC5载体,转染C3H10T1/2细胞,采用qRT-PCR检测成脂分化关键基因的表达情况,油红O染色检测脂滴含量,利用Western印迹检测细胞外信号调节激酶(extracellularregulatedkinase1/2,ERK1/2)及磷酸化ERK1/2(P-ERK1/2)的表达水平。C3H10T1/2细胞成脂诱导分化8d,Fndc5的表达量明显升高(P<0.05);C3H10T1/2细胞中过表达FNDC5,成脂分化关键基因过氧化物酶体增殖物激活受体-γ(peroxisome proliferator-activated receptor-γ,PPARγ)、脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)和CCAAT增强子结合蛋白α(CCAAT enhancer binding protein alpha, C/EBPα)的表达量显著降低(P<0.01),CCAAT增强子结合蛋白β(CCAAT enhancer binding protein beta, C/EBPβ)表达量明显降低(P<0.05),脂滴含量明显减少,P-ERK1/2的含量明显降低(P<0.05)。C3H10T1/2细胞中干扰FNDC5,成脂分化关键基因PPARγ、C/EBPβ、FABP4和C/EBPα的表达量显著升高(P<0.01),脂滴含量明显增加,P-ERK1/2的含量明显升高(P<0.05)。本研究发现,FNDC5可以通过抑制ERK1/2的磷酸化水平,抑制C3H10T1/2细胞的成脂分化,为FNDC5调控脂肪沉积的机制研究提供参考数据。The aim of this study was to explore the regulatory mechanism of Type Ⅲ domain-containing protein5(FNDC5) on adipogenic differentiation in C3 H10 T1/2 cells. qRT-PCR and Western blot were used to detect the expression of FNDC5 during adipogenic differentiation of C3 H10 T1/2 cells. The lentivirus-coated overexpression and interference vector of FNDC5 were constructed and transfected into C3 H10 T1/2 cells. qRT-PCR was used to detect the expression of the key genes of adipogenic differentiation. Oil red O staining was used to detect the formation of lipid droplets;Western blot was used to detect the content of ERK1/2 and ERK1/2 phosphorylated protein(P-ERK1/2). After 8 days of adipogenic differentiation of C3 H10 T1/2 cells, the expression of Fndc5 increased significantly. After overexpression of FNDC5 in C3 H10 T1/2 cells, the expression of key genes for adipogenic differentiation, including peroxisome proliferator-activated receptor-γ(PPARγ), CCAAT enhancer binding protein beta(C/EBPβ), fatty acid binding protein 4(FABP4) and CCAAT enhancer binding protein alpha(C/EBPα), all decreased significantly. The content of lipid droplets and P-ERK1/2 also decreased significantly. On the contrary, after interference of FNDC5 in C3 H10 T1/2 cells, the expression of key genes for adipogenic differentiation, including PPARγ, C/EBPβ, FABP4 and C/EBPα were significantly increased. Meanwhile, the content of lipid droplets and P-ERK1/2 also increased significantly. This study found that FNDC5 can inhibit the adipogenic differentiation of C3 H10 T1/2 cells by inhibiting the phosphorylation level of ERK1/2, which can provide reference data for the mechanism of FNDC5 in regulating fat deposition.

关 键 词：C3H10T1/2细胞 成脂分化 Ⅲ型纤连蛋白组件包含蛋白5基因 ERK1/2 

分 类 号：Q291[生物学—细胞生物学]