3T3-L1前脂肪细胞中Ghrelin对miRNAs表达的影响

为探讨3T3-L1前脂肪细胞增殖分化过程中Ghrelin对microRNAs(miRNAs)表达的影响,将pcDNA3.1(+)-Ghrelin转染至3T3-L1前脂肪细胞,real-time PCR检测10种miRNA的表达变化,并对显著差异表达的miRNA进行生物信息学分析。结果显示,pcDNA3.1(+)-Ghrelin的表达明显上调,约是对照组的5倍;real-time PCR结果显示,miR-125b-5p(P<0.05)表达显著上调,miR-21a-5p(P<0.01)、miR-30c-5p(P<0.05)、miR-181-5p(P<0.01)表达显著下调;生物信息学分析结果显示,预测调节的1062个靶基因与细胞的生长、分化和发育功能相关,核因子1b(Nfib)是其共同调节的重要潜在靶基因;丝裂原活化蛋白激酶(MAPK signaling pathway)和成脂基因信号通路是其调节差异最为显著的信号通路,与脂肪细胞的增殖和分化密切相关。结果表明,Ghrelin可通过调节miRNA的表达影响3T3-L1前脂肪细胞的增殖和分化。研究结果为进一步探讨Ghrelin发挥效应的分子调节机制和分子调控网络奠定了基础。

Effect of Ghrelin on miRNAs Expression in 3T3-L1 Preadipocytes

To investigate the effect of ghrelin on the expression of microRNAs(miRNAs)during the proliferation and differentiation of 3T3-L1 preadipocytes,pcDNA3.1(+)-Ghrelin was transfected into 3T3-L1 preadipocytes,and the expressions of 10 miRNAs were detected by qPCR,the differentially expressed miRNAs were analyzed by bioinformatics.The results showed that the expression of pcDNA3.1(+)-Ghrelin was significantly up-regulated,about five times higher than that of the control group;the qPCR results showed that the expression of miR-125b-5p(P<0.05)was significantly up-regulated,and the expression of miR-21a-5p(P<0.01),miR-30c-5p(P<0.05)and miR-181-5p(P<0.01)were significantly down-regulated;the bioinformatics analysis showed that the 1062 target genes predicted for regulation were related to the cell growth,differentiation and developmental functions,and nuclear factor 1 b-type(Nfib)was a potential important target gene for its co-regulation.Both the MAPK and adipogenesis gene pathways are the most significant signaling pathways that regulated by miRNAs,which are closely related to adipocyte proliferation and differentiation.These results suggest that Ghrelin can affect the proliferation and differentiation of 3T3-L1 preadipocytes by regulating the expression of miRNAs.This study laid a foundation for further exploring the molecular regulatory mechanism and network of Ghrelin effect.