黄芩苷诱导大鼠骨髓间充质干细胞体外分化为胰岛素分泌细胞  

Transdifferentiation of rat bone marrow mesenchymal stem cells into islet-secreting cells by baicalin 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肾病研究所,210002 [2]中南大学湘雅医院 [3]郑州大学第一附属医院

出　　处：《中华实验外科杂志》2007年第4期413-414,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(30200128);中国博士后科学基金(20060390283)

摘　　要：目的体外诱导大鼠骨髓间质干细胞(BMSCs)分化为胰岛样细胞。方法采用分步法体外诱导后,间接免疫荧光法鉴定诱导前后细胞nestin、胰岛素蛋白表达;RT-PCR法检测诱导前后胰岛转录因子mRNA表达;EHSA检测诱导后细胞葡萄糖刺激的胰岛素分泌。结果免疫荧光染色显示诱导5 h,nestin阳性细胞为(44.6±7.3)%。诱导24 h,nestin阳性细胞增至(61.8±8.4)%。此后,nestin阳性细胞数目开始下降,诱导第14天后,nestin表达基本消失;同时诱导后的细胞可以表达胰岛素蛋白。另一方面,RT-PCR结果显示诱导后细胞可以表达胰岛素-1、葡萄糖转运子-2及其转录因子mRNA。ELISA结果显示不同浓度的葡萄糖刺激的胰岛素分泌量不同,5 mmol/L和25mmol/L葡萄糖刺激的胰岛素分泌量分别为(25.53±6.49)和(53.26±7.56)mU/L,而诱导前MSCs不具备上述特点。结论大鼠BMSCs体外可以诱导成为胰岛素分泌细胞。Objective To induce the differentiation of marrow mesenchymal stem cells （BMSCs） into islet-secreting cells in vitro. Methods Using a defined culture medium and technique for transdifferentiation, BMSCs from adult SD rats were guided into specific insulin- secreting cells. The expression of insulin was detected by indirect immunofluorescence cytoehemistry staining before and after induction. And the expression of pancreatic islet cell differentiation-related transcripts, such as insulin Ⅰ, glucose transporter2 （GLUT2）,Isl-1 ,Pdx-1 ,Pax-4 and Pax-6 ,was detected by reverse transcription-PCR （RT-PCR）. In addition,insulin secretion was examined using EL/SA. Results After induction for 5 h, （44.6 ±7.3 ） % differentiated BMSCs expressed nestin, a marker of neural stem cells, （ 61.8 ± 8.4） % at 24 h, but 0 at day 14. Meantime, islet-like cellular clusters appeared after day 14 and became more Apparent by day 28. Differentiated cells were found to be immtmoreactive to insulin and expressed insulin 1, GLUT2 mRNA and other islet-associated transcription facrors. Glucose-induced insulin secretion in 5 and 25 mmol/L glucose was （25.53 ± 6.49） and （ 53.26 ±7.56） U/mL respectively. Conclusion Adult rat BMSCs can be differentiated into insulin-secreting cells in vitro.

关 键 词：骨髓 间充质干细胞 胰岛 糖尿病 

分 类 号：R285.5[医药卫生—中药学]