Ghrelin对鸡脂肪间充质干细胞增殖及分化为脂肪细胞的影响

Effects of ghrelin on the proliferation and differentiation of chicken adipose-derived mesenchymal stem cells (AMSCs)

为探讨Ghrelin对鸡脂肪间充质干细胞(AMSCs)增殖及分化为脂肪细胞的影响。本研究采用CCK-8法检测不同浓度的Ghrelin对AMSCs增殖的影响,再通过实时荧光定量PCR检测Ghrelin对AMSCs中c-myc和胸苷激酶1(TK1)基因mRNA表达水平的影响。然后,采用化学法对AMSCs进行成脂分化诱导,在此过程中添加不同浓度的Ghrelin,观察AMSCs的形态学变化,油红染色测定甘油三酯的累积情况,并通过实时荧光定量PCR检测脂肪细胞分化转录因子过氧化物酶体增殖剂活化受体γ(PPARγ)和CAAT/增强子结合蛋白α(C/EBPα)基因mRNA表达水平的变化。结果显示,10-7~10-11 mol/L浓度的Ghrelin均能显著或极显著促进AMSCs的增殖,其中10-9 mol/L浓度的Ghrelin的促增殖作用最强;Ghrelin也能显著或极显著升高c-myc和TK1基因mRNA的表达量。同时,Ghrelin促进AMSCs分化为脂肪细胞过程中甘油三酯的累积和脂滴的形成,显著或极显著升高PPARγ和C/EBPα基因mRNA的表达水平。结果说明,Ghrelin能够促进AMSCs增殖及分化为脂肪细胞。其分子调节机制可能是,Ghrelin通过增加c-myc的含量,进而引起TK1的活化,从而导致细胞周期的激活,促进AMSCs增殖;Ghrelin可能通过促进PPARγ和C/EBPα的表达,从而促进AMSCs分化为脂肪细胞。

Abstract：To explore the effects of ghrelin on the AMSCs proliferating and differentiating into adi- pocytes. CCK-8 and qPCR were used to detect the proliferation potentials of AMSCs treated with different concentrations of Ghrelin and the mRNA expression levels of c-myc and thymidine kinase 1 gene;and during adipogenic differentiation of AMSCs via chemically induction,the change of cell morphology, the accumulation of triglycerides and the mRNA expression levels of peroxisome pro- liferator-activated receptor gamma （PPARγ） and CCAAT enhancer binding protein alpha （C/ EBPα） were detected. 10-7- 10 11 mol/L of Ghrelin can significantly promote AMSCs prolifera- tion,especially 10 9 mol/L, qPCR demonstrated that the mRNA expression levels of c-myc and thymidine kinase 1 gene significantly increased in ghrelin group, ghrelin can also promote the accu- mulation of triglycerides and the formation of lipid drops in the process of the AMSCs differentia- ting into adipocytes,and the mRNA expression levels of PPARγ and C/EBPα significantly in- creased in ghrelin group. Results showed that ghrelin can promote proliferation of AMSCs via en- hancing expression of c-myc and activing thymidine kinase,then leading to activation of the cell cy- cle. Ghrelin can also promote the differentiation of AMSCs into adipocytes by increasing expression of PPARγ and C/EBPα.