心脉隆注射液诱导大鼠骨髓间充质干细胞分化为神经元样细胞的研究

山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (7): 32-36.

心脉隆注射液诱导大鼠骨髓间充质干细胞分化为神经元样细胞的研究

罗珊珊1，于君2，王晓云1，刘震1，满潇潇1，谢兆宏1，毕建忠1

1.山东大学第二医院神经内科, 济南 250033； 2.山东中医药大学附属医院，济南 250011

收稿日期:2012-03-08 出版日期:2012-07-10 发布日期:2012-07-10
通讯作者: 毕建忠(1961- )，男，博士，教授，博士生导师，主要从事老年性痴呆及脑血管病研究。 E-mail:bjz@sdu.edu.cn
作者简介:罗珊珊(1986- )，女，硕士研究生，主要从事老年性痴呆研究。 E-mail:xmluoshanshan@126.com
基金资助:
国家自然科学基金(81171214)；山东省优秀中青年科学家科研奖励基金(BS2009SW054)。

Differentiation of rat bone marrow mesenchymal stem cells into neuron like cells induced by Xinmailong injection

LUO Shan-shan1, YU Jun2, WANG Xiao-yun1, LIU Zhen1, MAN Xiao-xiao1, XIE Zhao-hong1, BI Jian-zhong1

1. Department of Neurology, Second Hospital of Shandong University, Jinan 250033, China;
2.The Affiliated Hospital of Shandong Traditional Chinese Medicine University, Jinan 250011, China

Received:2012-03-08 Online:2012-07-10 Published:2012-07-10

摘要/Abstract

摘要：

目的探讨心脉隆注射液体外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。方法采用贴壁培养法分离纯化大鼠骨髓间充质干细胞，流式细胞仪检测细胞表面抗原的表达，免疫细胞荧光法检测诱导后巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)、微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP)的表达。结果流式细胞仪检测显示CD29、CD90阳性，CD34、CD45阴性。实验组诱导4、12h后Nestin阳性表达率分别为(81.0±1.6)%、(22.5±1.9)%，36h后Nestin阴性；诱导4h后NSE、MAP2阴性，12、36h后NSE阳性表达率分别为(73.5±2.2)%、(94.3±1.8)%，MAP2阳性表达率分别为(80.0±2.2)%、(96.4±2.8)%；诱导4、12、36h后GFAP阴性。结论心脉隆注射液可在体外快速、高效地诱导大鼠骨髓间充质干细胞向神经元样细胞分化。

关键词: 干细胞；细胞分化；神经元；大鼠；大蠊属

Abstract:

Objective To explore the feasibility of Xinmailong injection inducing rat bone marrow mesenchymal stem cells (BMSCs) to differentiate into neuronlike cells. Methods The rat BMSCs were separated and purified with the adherent culture method. Following passages, the cell surface markers CD29, CD90, CD34 and CD45 were measured by flow cytometry. The expressions of Nestin, Neuron specific enolase (NSE), Microtubule associated protein2(MAP2) and Glial fibrillary acidic protein (GFAP) were detected by immunofluorescence assay. Results Flow cytometry showed positive expression of CD29 (99.06%) and CD90 (99.61%) and negative expression of CD34 (2.03%) and CD45 (2.00%). At 4h, 12h after induction, the positive rates of Nestin were (81.0±1.6)%, (22.5±1.9)%, while at 36h after induction, Nestin was negative. At 4h after induction, NSE and MAP2 were negative. At 12h, 36h after induction, the positive rates of NSE were (73.5±2.2)%, (94.3±1.8)% and those of MAP2 were (80.0±2.2)%, (96.4±2.8)%. At 4h, 12h, 36h after induction, GFAP was nagative. Conclusion Xinmailong injection can rapidly and efficiently induce rat BMSCs to differentiate into neuron like cells.

Key words: Stem cells; Cell differentiation; Neurons; Rats; Periplaneta

中图分类号:

R741.05

罗珊珊1，于君2，王晓云1，刘震1，满潇潇1，谢兆宏1，毕建忠1. 心脉隆注射液诱导大鼠骨髓间充质干细胞分化为神经元样细胞的研究[J]. 山东大学学报(医学版), 2012, 50(7): 32-36.

LUO Shan-shan1, YU Jun2, WANG Xiao-yun1, LIU Zhen1, MAN Xiao-xiao1, XIE Zhao-hong1, BI Jian-zhong1. Differentiation of rat bone marrow mesenchymal stem cells into neuron like cells induced by Xinmailong injection[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(7): 32-36.

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