【摘要】 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。更多还原

【Abstract】 To explore the effects of ghrelin on the AMSCs proliferating and differentiating into adipocytes.CCK-8and 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 kinase1gene;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 proliferator-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 proliferation,especially 10-9 mol/L.qPCR demonstrated that the mRNA expression levels of c-myc and thymidine kinase 1gene significantly increased in ghrelin group.ghrelin can also promote the accumulation of triglycerides and the formation of lipid drops in the process of the AMSCs differentiating into adipocytes,and the mRNA expression levels of PPARγand C/EBPαsignificantly increased in ghrelin group.Results showed that ghrelin can promote proliferation of AMSCs via enhancing expression of c-myc and activing thymidine kinase,then leading to activation of the cell cycle.Ghrelin can also promote the differentiation of AMSCs into adipocytes by increasing expression of PPARγand C/EBPα.更多还原