PDX-1联合利拉鲁肽诱导大鼠骨髓间充质干细胞分化为胰岛样细胞的效果*
Effect of PDX-1 combined with liraglutide on differentiation of bone marrow mesenchymal stem cells into insulin-producing cells

目的:探讨胰十二指肠同源盒-1(PDX-1)联合利拉鲁肽诱导大鼠骨髓间充质干细胞(BMSCs)分化为胰岛样细胞(IPCs)的效果。方法:构建真核表达载体pEGFP-pcDNA3.1(+)-PDX-1,脂质体介导其转染贴壁培养法分离得到的大鼠BMSCs,用潮霉素筛选稳定转染细胞PDX-1-BMSCs。将BMSCs和PDX-1-BMSCs分别用50 nmol/L利拉鲁肽或培养液培养10 d,然后ELISA法检测细胞上清胰岛素浓度,real-time PCR法检测细胞中Tle、Nkx6.1、Ngn3、Nestin、PDX-1 mRNA的表达。结果:4组细胞诱导过程中形态上趋向胰岛样细胞分化; 利拉鲁肽和PDX-1均可诱导BMSCs胰岛素分泌水平增加(P<0.05); PDX-1可促进BMSCs中PDX-1、Ngn3、Nkx6.1、Tle mRNA的表达(P<0.05); 利拉鲁肽可促进PDX-1、Ngn3 mRNA的表达(P<0.05); 两者有协同作用的趋势。结论:PDX-1 联合利拉鲁肽可以诱导BMSCs向IPCs分化并分泌胰岛素。
Aim: To investigate the effect of pancreatic duodenal homeobox-1(PDX-1)and liraglutide on differentiation of bone marrow mesenchymal stem cells(BMSCs)into insulin-producing cells(IPCs).Methods: Eukaryotic expression vector pEGPF-pcDNA3.1(+)-PDX-1 was constructed, then transfected via liposome into rat BMSCs separated and purified by attachment culture method in vitro, and stable transfection cells PDX-1-BMSCs were screened by homomycin. PDX-1-BMSCs and BMSCs were cultured in the medium with or without 50 nmol/L liraglutide for 10 days. The secretion of insulin was measured by ELISA. The mRNA expressions of Tle, Nkx6.1,Ngn3,Nestin, and PDX-1 were detected by real-time PCR.Results: The morphology of induced cells trended to islet-like cluster cells in the differentiation process. PDX-1 transfection and liraglutide inducing could increase the secretion of insulin of BMSCs(P<0.05). PDX-1 transfection could increase the mRNA expressions of Tle, Nkx6.1,Ngn3, and PDX-1(P<0.05),and liraglutide inducing could increase the mRNA expressions of Ngn3 and PDX-1(P<0.05); PDX-1 transfection and liraglutide inducing had synergistic effect.Conclusion: PDX-1 combined with liraglutide could promote BMSCs differentiating into IPCs and secreting insulin