虎杖苷联合生长因子诱导人脐带间充质干细胞向神经元样细胞分化的研究  

Study on neuron-like cells differentiation of human umbilical cord mesenchymal stem cells induced by polydatin combination with growth factors

作　　者：阮玲娟[1,2] 崔戈 顾旭东 莫娟芬 傅建明 鲍轶 

机构地区：[1]嘉兴市第二医院,嘉兴314000 [2]浙江省安吉县人民医院,湖州313300

出　　处：《中华中医药杂志》2015年第1期284-287,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省中医药科学研究基金项目(No.2011ZA104);国家自然青年科学基金项目(No.811010707);嘉兴科技局项目(No.2012AY1071-2)~~

摘　　要：目的:探讨虎杖苷联合生长因子诱导人脐带间充质干细胞(h UC-MSC)向神经元样细胞分化的过程。方法:采用原代组织贴壁法,体外分离培养h UC-MSC,流式细胞术鉴定h UC-MSC表面标志物。不同剂量虎杖苷联合生长因子诱导后显微镜下观察细胞形态变化,应用荧光定量PCR方法检测神经元特异性标志物3型β微管蛋白(β-tubulin-Ⅲ)、微管相关蛋白2(MAP2)和孤束核受体相关因子1(Nurr1)的表达,细胞免疫荧光技术检测β-tubulin-Ⅲ的蛋白表达。结果:h UC-MSC高表达CD29、CD44、CD105,极低表达CD14、CD34、CD45、HLA-DR。诱导后部分h UC-MSC分化为典型的神经元样细胞,并且β-tubulin-Ⅲ、MAP2基因表达经诱导后显著上调。Nurr1 m RNA仅在高剂量虎杖苷联合生长因子组显著上调。不同剂量虎杖苷联合生长因子组均有部分细胞表达β-tubulin-Ⅲ,对照组几乎不表达。结论:虎杖苷能够诱导h UC-MSC分化为神经元样细胞,且存在剂量依赖性,为h UC-MSC移植治疗神经系统疾病提供了实验依据。Objective： To investigate the process of neuron-like cells differentiation of human umbilical cord mesenchymal stem cells （hUC-MSC） induced by polydatin combination with growth factors. Methods： The hUC-MSC were isolated and cultured in vitro with explant technique. The surface marker of hUC-MSC was determined by flow cytometry method. The cellular morphology induced by different dose polydafin combination with growth factors was observed under microscope. The qRT-PCR was used to detect the expression of Class Ⅲ beta tubulin （β -tubulin-Ⅲ） mRNA, microtubule-associated protein 2 （MAP2） mRNA, and orphan nuclear receptor correlation factor I （Nurrl） mRNA. Immunofluorescence technique was used to detect the expression of β -tubulin-Ⅲ protein. Results： It showed that CD29, CD44 and CD105 were overexpression and CD34, CD45 and HLA-DR were low expression on hUC-MSC. The part of hUC-MSC induced by polydatin combination with growth factors had differentiated typical neuron-like cells. The expression of β -tubulin-Ⅲ mRNA, MAP2 mRNA both showed obviously up-regulation, while the expression of Nurrl mRNA up-regulation induced by high dose polydatin combination with growth factors. The β -tubulin- Ⅲ protein were expressed on a part of hUC-MSCs induced by different doses of polydatin combination with growth factors, while scarcely on the control group. Conclusion： The polydatin could induce neuron-like cells differentiation of hUC-MSC, the effect of which may depend on the dose of polydatin. To some extent, it could provide experimental basis for the treatment of nervous system diseases by hUC-MSC transplantation.

关 键 词：虎杖苷 生长因子 人脐带间充质干细胞 神经元样细胞 细胞分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]