胸腺嘧啶DNA糖基化酶基因敲低会影响猪前脂肪细胞的分化。

Thymine DNA Glycosylase Gene Knockdown Can Affect the Differentiation of Pig Preadipocytes.

作者信息

Zhang Lian-Jiang, Liu Si-Yuan, Zhu Ya-Nan, Gao Yan, Chen Jian, Yuan Bao, Jiang Hao, Dai Li-Sheng, Zhang Jia-Bao

机构信息

College of Animal Science and Technology, Jilin Agricultural Science and Technology College, Jilin, China.

出版信息

Cell Physiol Biochem. 2016;39(3):975-84. doi: 10.1159/000447805. Epub 2016 Aug 12.

DOI:10.1159/000447805

PMID:27513857

Abstract

AIMS

To study the effect of thymine DNA glycosylase (TDG) gene knockdown on the differentiation of pig preadipocytes.

METHODS

Preadipocytes were obtained from subcutaneous adipose tissue from the neck of 1- to 7-day-old pigs. The TDG gene was knocked down using siRNA, and cell differentiation was induced. The mRNA expression level was measured using fluorescence quantitative PCR, and the protein expression level was determined using Western blot analysis. The DNA methylation levels in promoter regions of differentiation-related genes were also evaluated.

RESULTS

TDG gene knockdown decreased the mRNA expression levels of the peroxisome proliferator-activated receptorγ (PPARγ) and Fatty acid binding proteins 4(FABP4 Also known as aP2) genes (P<0.01), while the mRNA expression level of the CCAAT/enhancer binding protein alpha(C/EBPα) gene did not change significantly (P>0.05). In addition, after induced differentiation, the lipid droplet production significantly decreased, and the percentages of methylation in the promoter regions of C/EBPα, PPARγ, and aP2 genes were 0.9%, 80%, and 76%, respectively. In contrast, the percentages of methylation in the negative control groups were 0.5%, 67.5%, and 58%, respectively.

CONCLUSION

TDG gene knockdown could inhibit the differentiation of pig preadipocytes and affect the DNA methylation levels of some transcription factors.

摘要

目的

研究胸腺嘧啶DNA糖基化酶（TDG）基因敲低对猪前体脂肪细胞分化的影响。

方法

从1至7日龄仔猪颈部皮下脂肪组织获取前体脂肪细胞。使用小干扰RNA（siRNA）敲低TDG基因，并诱导细胞分化。采用荧光定量PCR检测mRNA表达水平，利用蛋白质免疫印迹分析测定蛋白质表达水平。同时评估分化相关基因启动子区域的DNA甲基化水平。

结果

TDG基因敲低使过氧化物酶体增殖物激活受体γ（PPARγ）和脂肪酸结合蛋白4（FABP4，也称为aP2）基因的mRNA表达水平降低（P<0.01），而CCAAT/增强子结合蛋白α（C/EBPα）基因的mRNA表达水平无显著变化（P>0.05）。此外，诱导分化后，脂滴生成显著减少，C/EBPα、PPARγ和aP2基因启动子区域的甲基化百分比分别为0.9%、80%和76%。相比之下，阴性对照组的甲基化百分比分别为0.5%、67.5%和58%。