bFGF联合香丹注射液诱导间充质干细胞向神经元样细胞分化  

The Effect on Differentiation and Functions of Rat Bone Marrow Mesenchymal Stem Cells Induced to Neuron-like Cells by bFGF and Xiangdan Injection

作　　者：陈丹丹[1] 付文玉[1] 庄文欣[2] 吕翠[3] 

机构地区：[1]潍坊医学院组织胚胎学教研室,山东潍坊261053 [2]潍坊医学院医学研究试验中心,山东潍坊261053 [3]潍坊医学院附属医院干细胞实验室,山东潍坊261053

出　　处：《辽宁医学院学报》2011年第3期213-216,I0002,共5页Journal of Liaoning Medical University (LNMU) Bimonthly

基　　金：山东省自然科学基金项目;编号:Y2003C01Y2008C129;留学回国人员科研启动基金;编号:教外司留[2009]1001号

摘　　要：目的研究碱性成纤维细胞生长因子(bFGF)联合香丹注射液诱导大鼠骨髓间充质干细胞(BMSCs)向神经元样细胞分化的可行性及分化后细胞功能的改变。方法体外分离、扩增和鉴定大鼠BMSCs;在使用Neurobasal medium培养的同时,采用bFGF预诱导2 d后,实施香丹注射液诱导3 d。免疫细胞化学方法鉴定诱导后细胞的性质,计数阳性细胞阳性率;荧光分析法测定诱导后细胞培养上清液中儿茶酚胺含量。结果经过诱导,大鼠BMSCs能够部分分化为神经元样细胞。诱导后免疫细胞化学染色呈神经巢蛋白(Nestin)、微管相关蛋白-Ⅱ(MAP-Ⅱ)、神经元特异性烯醇化酶(NSE)、酪氨酸羟化酶(TH)阳性,阳性表达率分别为(71.5±4.6)%,(43.05±9.6)%,(49.9±4.3)%和(30.4±5.3)%;SHH配体蛋白SMO在细胞中高表达,并且与对照组比较差异具有统计学意义。细胞上清液中儿茶酚胺类物质(CA)含量为(42.24±2.42)μg/L。结论 bFGF+香丹注射液组合能够诱导大鼠BMSCs分化为DA能神经元样细胞。Objective To investigate the differentiation feasibility of rat bone marrow mesenchymal stem cell （BMSCs） induced by basic fibroblast growth factor （bFGF） and Xiangdan Injection to neurons and the changes of the induced cells＇ function. Methods BMSCs derived from rat bone marrow were separated, amplified and identified in vitro. After being induced by bFGF for 2 days, BMSCs were incubated with bFGF and Xiangdan Injection for three days in Neurobasal medium. Then the cell properties were detected by immunocytochemical methods. The content of catecholamine （CA） in supernatant fluid was determined by fluorescent analysis, respectively. Results Under the inverted phase contrast microscope, the induced BMSCs in this combination showed typical neuronal morphological characteristics after induction. The expression rates of Nestin, microtubule - associated protein - Ⅱ （ MAP -Ⅱ） , neuron - specific enolase （ NSE ） and Tyrosine hydroxylase （TH） were （71.5±4.6） %, （43.05 ±9.6） %, （49.9 ±4. 3） % and （30. 4 ±5. 3） % respectively. The expression of Smoothened （SMO） of the cells induced with bFGF and Xiangdan Injection was higher than the normal cells （P 〈 0. 01 ）. The content of catecholamine （CA） in snpernatant fluid was （42. 24 ± 2.42） μg/L induced with bFGF and Xiangdan Injection. Conclusion bFGF and Xiangdan Injection could induce rat BMSCs to neuron -like cells effectively.

关 键 词：间充质干细胞 神经元 细胞分化 大鼠 

分 类 号：R321.5[医药卫生—人体解剖和组织胚胎学]