体外诱导骨髓基质干细胞治疗帕金森病大鼠模型的研究  

Study on bone marrow stromal cells induced in vitro to treat rat models of Parkinson’s disease

作　　者：叶民[1] 陈生弟[1] 汪锡金[1] 陆国强[1] 梁梁[1] 徐洁懿[1] 

机构地区：[1]上海第二医科大学附属瑞金医院神经科,200025

出　　处：《中华神经科杂志》2005年第10期624-627,共4页Chinese Journal of Neurology

基　　金：国家科技部"973"计划资助项目(G1999054008);国家自然科学基金资助项目(39970263);上海市卫生系统百名跨世纪优秀学科带头人培养计划资助项目(97BR001)

摘　　要：目的探讨骨髓基质干细胞(bonemarrowstromalcells,BMSCs)诱导分化为巢蛋白(nestin)阳性的神经干细胞后移植治疗帕金森病动物模型的疗效及作用机制。方法将建模成功的帕金森病SD大鼠模型分为3组,将溴尿嘧啶(brdu)标记的诱导分化的BMSCs(每只鼠共移植6×105个细胞)、未诱导的BMSCs(每只鼠共移植6×105个细胞)和生理盐水(12μl)分别注入模型鼠右侧纹状体,在治疗后1～5个月分别观察阿朴吗啡诱导的旋转行为的变化,并进行脑冰冻切片荧光免疫组织化学鉴定(免疫荧光检测双标细胞)以及脑切片酪氨酸羟化酶(tyrosinhydroxylase,TH)免疫组织化学检测TH阳性细胞并计数。结果细胞移植治疗后1个月,BMSCs诱导组和未诱导组旋转次数开始明显少于生理盐水组[前两组转数分别从移植前的(15.2±2.5)r/min、(14.4±3.8)r/min降到治疗后第5个月的(6.7±1.8)r/min和(8.5±3.2)r/min];而生理盐水组转数移植前后变化不明显,与前两组相比,差异有统计学意义(P<0.05);在移植治疗后,BMSCs诱导组移植区可见一定数量双标的神经和神经胶质细胞,而未发现明显的TH双标的细胞。通过对移植后TH阳性细胞的定量分析可发现:各组间黑质损毁侧TH阳性细胞生存率差异无统计学意义(P>0.05)。结论BMSCs诱导组治疗帕金森病模型的疗效总体好于未诱导的BMSCs组,前两组的疗效均好于生理盐水组。Objective To study curative effects and mechanism of bone marrow stromal cells （BMSCs） induced into nestin-positive cells transplantation on rat Parkinson＇s disease （PD） model. Methods The 6-hydroxydopamine （6-OHDA）-induced hemiparkinsonian rats were selected and allocated randomly into 3 groups. BMSCs with brdu-labeled induced into nestin positive cells （6 × 10^5cells/each model）, BMSCs without induced 6 × 10^5cells/each model and physiological saline （12 μl） were injected into right striatum of PD rat respectively. The rotation test was taken from the first month to the fifth month after transplantation. Brain frozen sections were carried out immunofluorescence double staining （brdu/ nestin, brdu/GFAP, brdu/NSE and brdu/TH ） after one week, one, three, five months of BMSCs transplantation, meanwhile immunohistochemical staining of the TH-immunoreactive （TH-IR） neurons and counting the number of TH-IR were done. Results Frequency of rotation was significantly less in induced or non-induced BMSCs groups than that in normal saline group between 1 and 5 months after transplantation （ Before transplantation, rotation times of induced and non-induced BMSCs groups were （ 15.2 ±2. 5 ） turns/ min, （14. 4 ±3.8） turns/min respectively, but after 5 months of transplantation rotation times of 2 groups were （6. 7 ± 1.8 ） turns/min and （8.5 ±3.2 ）turns/min ）;however, rotation times of normal saline group had no obviously change （ P 〈 0. 05 ）. Some brdu/nestin-positive cells could be found in the implanted region of BMSCs induction group at 1 week after transplantation, but disappeared a month later. At this time, brdu/GFAP and brdu/NSE-positive cells appeared except brdu/TH-positive cells. Between 1 and 5 months after transplantation, the number of TH-positive cells of the impaired side of the substantia nigra was not significantly different among all groups（ P 〉 0. 05 ）. Conclusions BMSCs induced in vitro had better effects on treating PD model rat than tha

关 键 词：间质干细胞移植 帕金森病 疾病模型 动物 

分 类 号：R742.5[医药卫生—神经病学与精神病学]