[1]闫淑芳,杜少斐,吴敬,等.高糖在联合转录因子诱导大鼠骨髓间充质 干细胞分化为胰岛样细胞中的作用[J].国际内分泌代谢杂志,2017,37(02):77-81.[doi:10.3760/cma.j.issn.1673-4157.2017.02.002]
　Yan Shufang*,Du Shaofei,Wu Jing,et al.Effects of high glucose on the differentiation of rat bone marrow mesenchymal stem cells into insulin-producing cells induced by transcriptional factors[J].International Journal of Endocrinology and Metabolism,2017,37(02):77-81.[doi:10.3760/cma.j.issn.1673-4157.2017.02.002]
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高糖在联合转录因子诱导大鼠骨髓间充质 干细胞分化为胰岛样细胞中的作用()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
37
期数:
2017年02期
页码:
77-81
栏目:
论著
出版日期:
2017-03-20

文章信息/Info

Title:
Effects of high glucose on the differentiation of rat bone marrow mesenchymal stem cells into insulin-producing cells induced by transcriptional factors
作者:
闫淑芳; 杜少斐; 吴敬; 刘宏霞; 袁慧娟
475000 开封,河南大学淮河医院内分泌科(闫淑芳); 450004 郑州市第一人民医院内分泌科(杜少斐); 476100 商丘市中心医院内分泌科(吴敬); 450003 郑州大学人民医院内分泌科(刘宏霞,袁慧娟)
Author(s):
Yan Shufang*;  Du Shaofei;  Wu Jing;  Liu Hongxia;  Yuan Huijuan.
*Department of Endocrinology, Huaihe Hospital of Henan University, Kaifeng 475000, China
关键词:
神经元素3;  胰-十二指肠同源盒因子-1;  高糖;  骨髓间充质干细胞;  胰岛样细胞
Keywords:
Neurogenin 3;  Pancreatic duodenal homeobox-1;  High glucose;  Bone marrow mesenchymal stem cell;  Insulin-producing cell
DOI:
10.3760/cma.j.issn.1673-4157.2017.02.002
摘要:
目的 探讨高糖在胰-十二指肠同源盒因子-1(Pdx-1)、神经元素3(Ngn 3)诱导大鼠骨髓间充质干细胞(BMSC)分化为胰岛样细胞(IPC)中的作用。方法 构建真核表达载体pcDNA3.1(+)/(zeo)-Ngn 3、pcDNA3.1(+)-Pdx-1,贴壁培养法分离Sprauge-Dawley雄性大鼠BMSC,流式细胞术鉴定BMSC,脂质体介导转染BMSC,并筛选稳定转染细胞培养于高糖(4.5 g/L)DMEM培养液,RT-PCR检测Ngn 3、巢蛋白、Pdx-1 mRNA的表达,ELISA法检测胰岛素浓度。结果 分离的细胞表面高表达CD44、CD105,而低表达CD34,符合BMSC特征。诱导过程中BMSC形态上趋向IPC分化,高糖培养组较低糖培养组细胞上清胰岛素浓度明显提高(F=25.77,P<0.05),高糖质粒培养组Pdx-1、Ngn 3及巢蛋白mRNA表达水平明显高于高糖培养组及质粒转染组(F=35.12、12.76、8.34, P均<0.05)。结论 高糖对Pdx-1联合Ngn 3诱导BMSC向IPC分化具有促进作用。
Abstract:
Objective To investigate the effects of high glucose on the differentiation of rat bone marrow mesenchymal stem cells(BMSC)into insulin-producing cells(IPC)induced by pancreatic duodenal homeobox-1(Pdx-1)and neurogenin 3(Ngn 3). Methods Eukaryotic expression vector of pcDNA3.1(+)/(zeo)-Ngn 3 and pcDNA3.1(+)-Pdx-1 were constructed. BMSC was separated and purified by attachment culture method in vitro. Pdx-1 and Ngn 3 were transfected to BMSC via Liposome. The stablely transfected cells were cultured in medium containing high glucose(4.5 g/L). The mRNA expression of Pdx-1, Ngn 3, Nestin were detected by RT-PCR. The secretion of insulin were measured by ELISA. Results The cell surface expressed high level of CD44 and CD105, but low level of CD34, which consistent with the characteristics of BMSC. Morphologically, BMSC had the trend to differentiate into islet cell during the induction process, and the level of insulin was higher in cells cultured in high glucose than in low glucose(F=25.77, P<0.05). The mRNA expression level of Pdx-1, Ngn 3, Nestin were increased in high glucose combined with liposome group than in high glucose group and liposome group(F=35.12, 12.76, 8.34, all P<0.05). Conclusion High glucose improves the differentition of BMSC into IPC induced by Pdx-1 and Ngn 3.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(U1204805) 通信作者:袁慧娟,Email: lmls3712@163.com
更新日期/Last Update: 2017-03-20