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

作　　者：闫淑芳[1] 吴敬[1] 刘宏霞[1] 杜少斐 张会峰[1] 史晓阳[1] 袁倩[1] 袁慧娟[1] 

机构地区：[1]郑州大学人民医院内分泌科,郑州450003

出　　处：《郑州大学学报（医学版）》2016年第2期161-166,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目U1204805

摘　　要：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p EGFP-pc DNA3.1(+)-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 differentia-tion of bone marrow mesenchymal stem cells （BMSCs） into insulin-producing cells（IPCs）.Methods：Eukaryotic expres-sion 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 in-crease 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 .

关 键 词：胰十二指肠同源盒-1 利拉鲁肽 大鼠 骨髓间充质干细胞 胰岛样细胞 

分 类 号：R587[医药卫生—内分泌]