摘要
目的探讨骨髓间充质干细胞（BMSCs）移植治疗脑梗死大鼠后梗死灶周围及室管膜下区神经元增殖与大鼠神经功能恢复的关系。方法采用密度梯度离心法分离、贴壁培养法培养大鼠BMSCs，并进行流式细胞仪鉴定。24只SD大鼠应用电凝法制作大脑中动脉闭塞（MCAO）模型成功后按随机数字表法分为对照组及移植组（每组各12只），其中移植组经静脉注射Hoechst33342标记BMSCs（浓度为5×10^6个细胞／μL），对照组注入lmL无血清DMEM／F12。BMSCs移植前、移植后1周、2周时采用横木行走试验评价2组大鼠神经功能状况。移植后1周、2周时脑组织取材，选择梗死灶周围及室管膜下区脑组织切片，荧光显微镜下观察BMSCs存活及分布情况，免疫组织化学染色检测Kj67阳性细胞的表达。结果移植后1周、2周时，移植组大鼠横木行走试验评分中位数分别为5、7，对照组大鼠分别为3、5，比较差异均有统计学意义（P〈0．05）。移植后1周、2周时，移植组大鼠梗死灶周围可见Hoechst33342荧光染色阳性细胞[分别为（7．78±1．19）个／视野、（8．15±0．90）个／视野]；同时梗死灶周围及室管膜下区Ki67阳性细胞数[（15．01±1．58）个／视野、（13．26±1．10）个／视野；（42．57±3．11）个／视野、（39．70±1．87）个／视野]明显多于对照组[（6．29±0．83）个／视野、（6．21±0．75）个／视野；（23．22±1．08）个／视野、（21．82±1．78）个／视野]，差异均有统计学意义（P〈0．05）。结论静脉移植BMSCs治疗脑梗死大鼠能促进大鼠神经功能恢复，其机制可能主要与其促进内源性神经元增殖有关。
Objective To explore the relation between neurologic function recovery and neurogenesis in subventricular zone （SVZ） and perinfarct area after bone marrow stromal cells （BMSCs） transplantation in stroke rats. Methods BMSCs were harvested from rats by density gradient centrifugation and adherent culture methods, and identified by flow cytometry. Middle cerebral artery occlusion （MCAO） rat models were established by electric coagulation method. They were randomly devided into control group and transplantation group （n=12）. The rats in the transplantation group were injected by tail vein with BMSCs （5×106 cells/μL） labeled with Hoechst 33342. Rats in the control group were injected with 1 mL of serum flee DMEM/F12. The functional recovery of rats in two groups was evaluated by using beam walking test before transplanation and i week and 2 weeks after BMSCs transplanation. The survival and distributions of transplated BMSCs in the subventricular zone （SVZ） and in peri-infarct area were observed by fluorescence microscope. Ki67 expression in SVZ and peri-infarctarea was determined by immunohistochemisty. Results Scores of beam walking test in transplantation group （median are 5 and 7 one and two weeks after transplantations, respectively） were significantly higher than thoses in control group （median are 3 and 5, respectively, P〈0.05）. One week and two weeks after transplantations, Hoechst positive cells were found in the peri-infarct area of rats in transplantation group （7.78±1.19 cell/field and 8.15±0.90 cell/field, respectively）; the number of Ki-67 positive cells in the peri-infarct area and SVZ of rats in transplantation group （15.01.19±1.58 cell/field and 13.26±1.10 cell/field; 42.57±3.11 cell/field and 39.70± 1.87 cell/field） was significantly increased as compared with those in the control group （6.29±0.83 cell/field and 6.21±0.75 cell/field; 23.22±1.08 cell/field and 21.82± 1.78 cell/field, P〈0.05）. Conclusions Neurogenesis in SVZ and perinfarct reg
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2014年第4期348-352,共5页
Chinese Journal of Neuromedicine
基金
广东省科技厅资助项目（20098030801354）
广州市教育局基金（61092）
浙江省医学会临床科研基金（2013zyc-A146）
番禺区科信局课题（2013-z-03-44）
关键词
脑梗死
骨髓间充质干细胞
KI67
细胞移植
神经功能
Cerebral infarction
Mesenchymal stem cell
Ki67
Cell transplantation
Neurological function
