脑室移植骨髓基质细胞对脑缺血损伤后神经功能恢复的影响  

Improvement of neurological functional recovery following bone marrow stromal cells transplantation to brain ventricle after cerebral ischemia

作　　者：王爽[1] 郑殿魁[2] 孙锋[2] 魏敏杰[1] 张朝东[3] 

机构地区：[1]中国医科大学药理学教研室,辽宁省沈阳市110001 [2]沈阳市第五人民医院神经内科,辽宁省沈阳市110023 [3]中国医科大学附属第一医院神经内科,辽宁省沈阳市110001

出　　处：《中国临床康复》2006年第21期27-30,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:采用脑室系统注入体外培养的骨髓基质细胞,观察其对神经功能损伤修复的影响及其在脑内的存活、迁移、分化情况。方法:实验于2004-03/12在中国医科大学神经内科实验室完成。①将28只Wistar大鼠随机分为假手术组(n=6),模型组(n=8),梗死半球移植组(n=7),健侧半球移植组(n=7)。②除假手术组外,其他3组大鼠采取longa法制备大脑中动脉梗死模型,梗死后次日将培养的骨髓基质细胞悬液(10μL,约1×105个细胞)注入梗死半球移植组和健侧半球移植组大鼠侧脑室,模型组注入等量的磷酸盐缓冲液。③通过神经功能评分观察大鼠脑神经功能恢复情况,采用免疫组织化学方法观察移植入脑的骨髓基质细胞的存活、迁移及分化情况。结果:28只大鼠全部进入结果分析。①神经功能缺损评价:假手术组动物无神经功能缺损。移植前各组梗死后大鼠神经功能损害评分差异无显著性(P>0.05),移植2周后梗死大鼠神经功能均有不同程度恢复,但梗死半球移植组、健侧半球移植组评分明显低于模型组(6.71±0.49,6.86±0.38,8.63±0.52,P<0.05)。②大鼠脑病理改变:苏木精-伊红染色显示大脑中动脉梗死大鼠的缺血区脑组织稀疏,细胞数量明显减少且间隙增大。假手术组则无此表现,表明大鼠脑梗死模型制作成功。③脑组织切片免疫组织化学染色结果:可见来源于骨髓基质细胞的细胞(5-溴脱氧尿嘧啶核苷阳性细胞经免疫组织化学染色呈暗棕色)在脑组织内存活,并且聚集于缺血梗死区域。在梗死半球移植组对侧半球脑组织内未发现5-溴脱氧尿嘧啶核苷阳性细胞;在健侧半球移植组仅在移植损伤道周围发现少量5-溴脱氧尿嘧啶核苷阳性细胞。应用免疫双标染色随机选取10个5-溴脱氧尿嘧啶核苷阳性细胞区域计数发现大鼠脑梗死后14d约1.2%的5-溴脱氧尿嘧啶核苷阳性细胞同时表达神经元标志蛋白特异性烯醇AIM： To observe the effect of neurological functional recovery after transplanting the cultured in vitro bone marrow stromal cells （BMSCs） into brain ventricle, and study the survival, migration and differentiation of BMSCs. METHODS： The experiment was conducted at the Neurological Laboratory of China Medical University between March and December 2004. ① Twenty-eight Wistar rats were randomized into sham group （n=6）, model group （n=8）, infarcted hemisphere transplantation group （infarcted group, n=7） and uninjured hemisphere transplantation group （uninjured group, n=7）. ②Middle cerebral artery occlusion （MCAO） models were prepared with longa method in other 3 groups except sham group. Next day after infarction, the cultured BMSCs suspension （10μL containing about 1×10^5 cells） was injected into the lateral ventricles of rats in infarcted group and uninjured group, while phosphate buffer of equal dosage was given in modal group. ③The recovery of neurological function in rats was assayed through neurological severity score （NSS）, and immunohistochemical method was applied to observe the survival, migration and differentiation of transplanted BMSCs in varied brain areas. RESULTS： Totally 28 rats were involved in the result analysis.①Evaluation of neurological impairment： No neurological impairment was found in rats of sham group. And there was no significant difference in NSS scores of all the infarcted rats before transplantation （P 〉 0.05）. Two weeks later, the neurological function of rats all recovered to some degrees, and rats transplanted with BMSCs had significantly lower NSS scores compared with rats of control group （6.71±0.49, 6.86±0.38, 8.63±0.52, P 〈 0.05）.②Pathological changes of rats＇ brains： The result of hematoxylin-eosin staining displayed sparse cerebral tissues in ischemia area of MCAO rats, with the decreased amount of cells and enlarged interspace. No manifestation was found in sham group, which indicated the MCAO models

关 键 词：骨髓细胞 移植 脑缺血 侧脑室 

分 类 号：R457.7[医药卫生—治疗学]