S100a10基因对小鼠前体脂肪细胞白色和棕色成脂分化的影响

Effects of S100a10 on White and Brown Adipogenic Differentiation of Mice Preadipocytes

旨在用小鼠模型探究全基因组关联分析(GWAS)筛选到的西门塔尔牛大理石花纹评分性状正相关基因S100钙结合蛋白A10( S100A10 )对小鼠前体脂肪细胞成脂分化的影响。本研究以来源于C57BL/6品系小鼠腹股沟两侧白色脂肪组织的前体脂肪细胞为材料,体外进行白色/棕色成脂分化。通过siRNA干扰 S100a10 基因表达,通过油红O染色检测前体脂肪细胞分化效果,通过荧光定量PCR(qRT-PCR)、蛋白免疫印迹和细胞免疫荧光检测基因和蛋白表达变化。结果显示,敲低 S100a10 后,通过油红O染色发现脂滴明显变少;通过qRT-PCR检测白色成脂分化标志基因 Pparg 、 C/ebpa 、 Fabp4 、 Glut4 、 Adiponectin 、 Leptin 表达量显著下降( P <0.01);棕色成脂分化基因 Ucp1、Pgc1a、Fabp4、Elovl3、Cox7a 表达量显著下降( P <0.01)。敲低 S100a10 后,通过蛋白免疫印迹检测发现,FABP4和PPARG在白色成脂分化中表达量显著下降( P <0.01),通过细胞免疫荧光检测发现FABP4在白色成脂分化中表达量下降;敲低 S100a10 后通过蛋白免疫印迹和细胞免疫荧光检测发现,UCP1和FABP4在棕色成脂分化中表达量显著下降( P <0.01)。综上表明,敲低 S100a10 基因后抑制小鼠前体脂肪细胞的白色/棕色成脂分化,本研究将为探究 S100A10 基因对于肉牛肌内脂肪沉积的影响提供科学依据。

The objective of this study was to investigate the effects of the S100A10 gene , which was identified as a candidate gene regulating Simmental marbling score by genome-wide association study (GWAS),on adipogenic differentiation of mice preadipocytes. In this study,white preadipocytes were isolated from inguinal white adipose tissue of C57BL/6 mice and induced white/brown adipogenic differentiation in vitro . The expression of the S100a10 gene was interfered using siRNA. The differentiation of preadipocytes were detected by Oil Red O staining. Gene and protein expression levels were measured by qRT-PCR,Western blot and immunofluorescence. The results showed that lipid droplets decreased significantly after S100a10 knocking down. And qRT-PCR assay showed that the expression level of white adipocyte-specific genes Pparg,C/ebpa , Fabp4 , Glut4 , Adiponectin and Leptin were significantly down regulated in white adipogenic differentiation( P <0.01). The expression level of brown adipocyte-specific genes Ucp1,Pgc1a,Fabp4,Elovl3 and Cox7a were significantly decreased in brown adipogenic differentiation( P <0.01). Western blot result showed that the expression of FABP4 and PPARG were significantly decreased in white adipogenic differentiation( P <0.01),the immunofluorescence assay showed that the expression of FABP4 was decreased in white adipogenic differentiation. The protein expression of brown adipogenic differentiation specific genes FABP4 and UCP1 were obviously decreased in brown adipogenic differentiation( P <0.01). The results showed that knocking down S100A10 inhibited the white/brown adipogenic differentiation in mice preadipocytes. This study provides scientific basis for the further research of the effect of S100A10 on intramuscular fat deposition in beef cattle.