摘要
前期研究中采用双萤光素酶报告基因验证了mi R-124与脂解因子猪TNF-α之间的靶关系,以此为基础研究mi R-124是否影响猪皮下脂肪细胞的分化。采用mi R-124模拟物mimics和抑制物inhibitor分别转染猪脂肪前体细胞并诱导其分化成成熟脂肪细胞,检测细胞的聚脂情况,甘油及甘油三酯的含量变化,荧光定量检测脂肪细胞关键转录因子PPARγ,脂肪合成和分解的主要酶FASN和HSL基因的表达变化。结果显示,过表达mi R-124能抑制TNF-α蛋白的表达,脂肪细胞脂滴多于对照组,甘油三酯(TG)含量显著增加(P<0.01),甘油含量亦显著增加(P<0.05),PPARγ、FASNT和HSL的表达显著上调(P<0.01);抑制细胞mi R-124的表达脂滴则较少,TG含量显著减少(P<0.05),PPARγ和FASN的表达均显著下调(P<0.05)。mi R-124可能通过抑制TNF-α调节猪脂肪细胞的分化,为后续研究mi R-124调节脂肪代谢的相关机制奠定基础。
In the previous research, porcine TNF-α was found to be the target of miR-124 by dual lueiferase assay. Here we verify whether miR-124 has the effect on the differentiation of porcine adipoeyte. Porcine pre-adipocyte was transfected with miR-124 mimics or inhibitor to over-express or to repress miR-124, and then induced into mature adipocytes. The accumulation of lipid droplets was observed using Oil Red O staining and the amount of glycerol and triglycerides （ TG ） were detected by TG assay and glycerol assay. The expressions of PPART （ proliferator-activated receptor-T ）, FASN （ fatty acid synthase ）, and HSL （ hormone-sensitive lipase ） were detected by real time fluorescence quantitative PCR. The result showed that overexpression of miR-124 repressed the expression of TNF-α protein, therefore, lipid droplets were more than that in the control, TG increased significantly （ P 〈 0.01 ）, also the same for glycerol （ P 〈 0.05 ）, and the expressions of PPART, FASNT, and HSL significantly up-regulated （P 〈 0.01 ） . Repressing miR-124 resulted in the reduction of lipid droplets and the significant decrease of TG, and the expressions of PPART and FASNT significantly down-regulated （ P 〈 0.05 ） . In conclusion, miR-124 might regulate the differentiation of porcine adipocyte by suppressing TNF-α, laying a foundation for future studies on the mechanism of miR-124 regulating lipid metabolism,
作者
李虹仪
李家标
郑艺林
吉卢琳
张茂
郑恩琴
LI Hong-yi LI Jla-biao ZHENG Yi-lin JI Lu-lin ZHANG Mao ZHENG En-qin(Animal Nutrition Scientific Research Innovation Team, College of Life Science, Longyan University, Longyan 364012 Guangdong Provincial Key Lab of Agro-animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642)
出处
《生物技术通报》
CAS
CSCD
北大核心
2017年第3期175-179,共5页
Biotechnology Bulletin
基金
福建省大学生创新创业训练计划项目(201511312055)
福建省自然科学基金项目(2014J05043)
龙岩学院博士启动基金项目(LB2013012)
