胰腺十二指肠同源框-1基因促进大鼠骨髓间充质干细胞体外横向分化为胰岛素分泌细胞  

Pancreatic duodenal homeobox-1 advances transdiffenentiation of bone marrow mesenchymal stem cells into insulin-producing cells in vitro

作　　者：孙吉平[1] 杨于嘉[2] 贾延劼[3] 

机构地区：[1]南京军区南京总医院解放军肾脏病研究所,江苏南京210002 [2]中南大学湘雅医院儿科,湖南长沙410008 [3]郑州大学第一附属医院神经内科,河南郑州450052

出　　处：《医学研究生学报》2006年第11期980-983,I0008-I0009,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助项目(批准号:30200128);中国博士后科学基金资助项目(批准号:20060390283)

摘　　要：目的:研究胰腺十二指肠同源框-1基因(Pdx-1)在骨髓间充质干细胞(MSC)体外分化中的作用。方法:构建含有Pdx-1基因的真核表达载体,转染介质Superfect介导重组载体转染MSC,G418筛选阳性细胞后对转染组(Pdx-1+MSC)和未转染组(Pdx-1-MSC)均体外进行诱导分化,细胞免疫组化染色鉴定诱导后胰岛素分泌阳性细胞。结果:限制性酶切分析和序列测定证实成功构建了含有Pdx-1基因的真核表达载体,荧光显微镜观察证实Superfect能高效介导重组载体转染MSC;细胞免疫组化染色显示Pdx-1+MSC分化为胰岛素分泌细胞的数量较Pdx-1-MSC明显增多,阳性率分别为(28.23±2.56)%和(7.08±2.69)%。结论:Pdx-1能增强大鼠MSC体外分化为功能性胰岛素分泌细胞的能力,为胰岛移植开辟新的思路,对1型糖尿病治疗有重要的应用价值。Objective： To investigate the possible effects of Pancreatic duodenal homeobox-1 （Pdx-1） expression in transdifferentiation of bone marrow mesenchymal stem cells （MSC） in vitro. Methods： The eukaryotic expression vector containing Pdx-1 was constructed. After such vector was transfected into MSC using Superfect, G418 was used to select the positive cells. Then both （ Pdx-1^＋ MSC ） and （Pdx-1^ -MSC） were induced to transdifferentiate in vitro. The expressions of insulin were analyzed by immunohistochemistry staining. Results： Restricted enzyme analysis and sequencing showed that interest gene segment was consistent with that in Gen Bank, Recombination vector was effectively transformed into MSC demonstrated by fluorescence microscope; insulin-producing cells from Pdx-1^＋ MSC were higher than that from Pdx-1 ^- MSC [ （28.23± 2.56 ） % and （ 7.08 ±2.69 ） %, respectively ]. Conclusion：Pdx-1 can promote adult rat MSC to transdifferentiate into insulin-producing cells in vitro,and this approach might lead to a widespread cell replacement therapy for type Ⅰ diabetes.

关 键 词：胰腺十二指肠同源框-1基因 骨髓间充质干细胞 胰岛 糖尿病 

分 类 号：Q784[生物学—分子生物学]