摘要
目的:从基因水平了解黄芪在诱导大鼠骨髓间充质干细胞向神经细胞分化过程中的作用途径。方法:实验于2002-09/2005-06在广州医学院病理生理学教研室和解剖教研室完成。①间充质干细胞的定向诱导分化:实验组加入预诱导液(20mL/L天麻注射液含体积分数为0.1的胎牛血清的L-DMEM),对照组只换液不加天麻。24h后吸去预诱导液,实验组加入诱导液(100mL/L黄芪注射液不含胎牛血清的L-DMEM),诱导细胞分化1h,对照组换液为不含胎牛血清的L-DMEM。②细胞总RNA的提取。③间充质干细胞诱导分化过程中差异表达基因的筛选。结果:①骨髓间充质干细胞及其诱导分化细胞:培养5～10代的细胞在黄芪诱导液中诱导1h后分化为神经元样细胞,免疫组织化学染色显示巢蛋白阳性,神经元特异性烯醇酶阳性,胶质纤维酸性蛋白阴性。②两组细胞的总RNA:紫外分光光度检查其吸光度(A值),A260/A280为1.8～2.0;甲醛变性凝胶电泳显示28S,18S,5S3条带。③在9052个基因中,与对照组相比,实验组检测到有379个基因表达下调,其中下调倍数大于2倍的有19个;表达上调的基因共有151个,其中有7个基因的上调倍数大于2,包括bHLH,epiregulin,fibroblastgrowthfactor9等基因。结论:黄芪可有效地诱导骨髓间充质干细胞分化神经元,其作用过程涉及多种基因。
AIM： To investigate the role of Astragale in the process of neural cell differentiation of rat mesonchymal stem cells from gene level.
METHODS： This experiment was carried out at the Department of Pathophysiology and Department of Anatomy of Guangzhou Medical College from September 2002 to June 2005. ① Directional induction and differentiation of mesenchymal stem cells： Pre-induction solution （20 mL gastredia rhizome in each liter L-DMEM parenteral solution containing 0.1 volume fraction of fetal bovine serum）, was added in experimental group. Solution was changed in control group, but gastredia rhizome was not added. Pre-induction solution was blotted 24 hours later. Another preinduction solution （100 mLAstragale in each liter L-DMEM not containing fetal bovine serum） was added to induce and differentiate into cells for one hour. L-DMEM without fetal bovine serum was given in control group. ② Extraction of cell total RNA. ③Screening of differential expression gene in the process of induction and differentiation of mesenchymal stem cells with Astragal, e and without Astragal, e by gene chip technology.
RESULTS： ① Mesonchymal stem cells and induced and differentiated cells： Astragal, e induced the cells of 5^th to 10^th generations to differentiate into neurons, immunohistochemistry showed nidogen and neuron specific enolase positive but glial fibrillary acidic protein negative. ②Total RNA of the cells in two groups ： Absorbance （A） was measured with a spectrophotometer, and results showed that A 260/A280 was about 1.8 to 2.0; 28S,18S,5S three bands appeared in the denaturing formaldehyde gel electrophoresis ③As compared with control group, among 9 052 rat genes, 530 genes had differential expression （up 151 genes, down 379 genes, including 〉 2 fold 7 and 〉 -2 fold 19）,including bHLH, epiregulin, fibroblast growth factor 9, etc.
CONCLUSION： Astragal, e can effectively induce mesenchymal stem cells to differentiate into neurons. The effect of Astragole during diff
出处
《中国临床康复》
CSCD
北大核心
2006年第21期1-3,共3页
Chinese Journal of Clinical Rehabilitation
基金
广东省自然科学基金资助项目(04009583)
广州市教育局基金资助项目(2004-1003)~~
