吉林大学学报(医学版) ›› 2013, Vol. 39 ›› Issue (1): 19-24.doi: 10.7694/jldxyxb20130106

• 基础研究 • 上一篇    下一篇

胎胰蛋白促进人脂肪来源的间充质干细胞向胰岛素及C-肽阳性细胞的分化及细胞鉴定

任莉莉1,陈丽娟1,齐晖1,李富荣2   

  1. (1.广东省深圳市人民医院临床医学研究中心,广东 深圳 518020;2.广东省深圳市老年医学研究所,广东 深圳 518020)
  • 收稿日期:2012-02-14 出版日期:2013-01-28 发布日期:2013-01-30
  • 作者简介:任莉莉(1975-),女,吉林省长春市人,副主任技师,免疫学博士,主要从事临床免疫学研究。
  • 基金资助:

    国家973计划前期研究专题资助课题(2007CB516811);广东省深圳市重点科技计划项目资助课题(200901001)

Protective effects of Gegencha on acute liver injury induced by acetaminophen in mice

REN Li-li1CHEN Li-juan1QI Hui1LI Fu-rong2   

  1. (1. Clinical Research Center,Shenzhen People’s Hospital,Guangdong Province,Shenzhen 518020,China;2. Shenzhen Institute of Geriatrics,Guangdong Province,Shenzhen 518020,China)
  • Received:2012-02-14 Online:2013-01-28 Published:2013-01-30

摘要: 目的:利用大鼠胚胎胰腺组织蛋白提取物诱导人脂肪来源的间充质干细胞(hASCs)向胰岛素分泌细胞分化,为临床胰岛移植寻找新的细胞来源。方法:将新生SD大鼠的胚胎胰腺匀浆后提取胎胰蛋白,重建胰腺微环境,诱导hASCs向胰腺方向分化。诱导组为胎胰蛋白培养组,对照组不添加胎胰蛋白,其余培养条件相同,诱导20 d后,荧光定量PCR技术检测2组胰腺发育相关基因的表达水平,ELISA法检测细胞悬液胰岛素水平,荧光免疫组织化学检测细胞C-肽蛋白表达。结果:在胎胰蛋白诱导过程中,hASCs重要标志基因Oct3/4及Nanog的表达持续降低,说明hASCs逐渐失去全能干细胞的特性;在诱导的第6天开始检测到PDX-1和CK-19的表达上调,诱导细胞出现了胰腺方向的分化,12 d开始检测到胰岛素的阳性表达。所获分化细胞基础相分泌胰岛素量为(287.17± 15.67) mIU•L-1,高糖刺激下为(401.09±28.94 )mIU•L-1,二者比较差异有统计学意义(P<0.01),且均明显高于对照组的(4.85±1.04) mIU•L-1 (P<0.01);免疫荧光检测,诱导组可见C肽阳性细胞,对照组未见C-肽阳性细胞。结论:模拟的微环境可诱导hASCs分化为有功能的胰岛素产生细胞,具有替代胰岛细胞治疗糖尿病的潜在可能性。

关键词: 间充质干细胞, 胰腺, 细胞分化, 细胞移植

Abstract: To use embryo pancreas extract (EPE) from rats to induce pancreatic differentiation of human adipose tissue-derived mesenchymal stem cells(hASCs) into insulin-secretion cells, and to find new cell source for islet transplantation in clinic.Methods  The pancreatic micro-circumstance was built for hASCs to differentiate into insulin-secretoin cells by extracting protein complex from new born SD rats.The hASCs in experiment group were induced by EPE and the cells in control group were cultured without EPE.20 d after induction,the expression levels of genes related with pancreas development  were detected by real-time PCR.ELISA  was used to detect the insulin secretion levels in experiment and control groups.The C-peptide expression was verified by fluorescent immunocytochemistry.Results The expressions of key markers of hASCs Oct 3/4 and Nanog were decreased in the process of EPE induction.On the 6th day after induction,the expression levels of POX-1 and CK-19 were up-regulated;and on the 12nd day,the insulin expression was positive.The insulin levels secreted by differentiated cells with and without glucose stimulation(401.09 mIU•L-1±28.94 mIU•L-1  vs   287.17 mIU•L-1± 15.67 mIU•L-1,P<0.01) were much higher than that of undifferentiated cells (4.85 mIU•L-1±1.04 mIU•L-1,P<0.01).The C-peptide positive cells were found in protein by fluorescent immunocytochemistry in experiment  group.Conclusion hASCs have the potential to differentiate into pancreas cell lineages with EPE induction.They could become substitution of islet cells in treatment of diabetes mellitus.

Key words: mesenchymal stem cells, pancreas, differentiation, cell transplantation

中图分类号: 

  • Q813.11