脂肪组织源性基质细胞经侧脑室立体定向移植治疗大鼠局灶性脑缺血的实验研究  

Stereotaxic transplantation of adipose tissue derived stromal cells into lateral cerebral ventricle for treatment of focal cerebral ischemia in rats

作　　者：周进[1] 田国萍 朱春艳[1] 吴丽华[1] 王静娥[1] 朱峰[1] 丛雪峰[1] 徐冰[1] 胡凤杰[1] 

机构地区：[1]辽宁省沈阳市第一人民医院沈阳脑科医院,沈阳110041

出　　处：《中华神经医学杂志》2006年第7期649-653,共5页Chinese Journal of Neuromedicine

基　　金：沈阳市科学技术计划基金(1053111-3-04)

摘　　要：目的探讨脂肪组织源性基质细胞(ADSCs)经侧脑室立体定向移植治疗大鼠局灶性脑缺血的机制。方法体外培养、扩增ADSCs,常规细胞免疫化学染色鉴定其种类；细胞移植前3d,加入含BrdU的培养液培养3d。缺血再灌注大鼠模型第4天经侧脑室移植ADSCs,于移植前、移植后第1、7、14、21、28天进行评分。移植后28 d,断头取脑,各组一部分脑片经红四氮唑(TTC)染色法检测脑梗死体积,另一部分切片用原位TUNEL法检测细胞凋亡；用免疫荧光双标法检测脑源性神经营养因子(BDNF)在有BrdU活性的ADSCs的表达。结果大多数ADSCs呈CD44(间质干细胞特征性标志物)阳性,CD114抗体(单核巨嗜细胞标志物)及CD34抗体(造血干细胞标志物)呈阴性。ADSCs组和对照组神经功能损害评分于脑缺血后21d、28d呈显著性差异(P＜0．01),脑梗死体积无显著性差异(P＜0．05)：免疫荧光双标法检测显示大部分BrdU标记的ADSCs迁移到梗死周边区,ADSCs组细胞凋亡数明显低于对照组(P＜0．01)。结论经侧脑室立体定向移植的ADSCs可能是通过减少梗死周边区细胞的凋亡促进大鼠局灶性脑缺血的神经功能恢复。Objective To explore the effect and mechanism of adipose tissue derived stromal cells （ADSCs） stereotaxically transplanted into the lateral cerebral ventricle for the treatment of focal cerebral ischemia in rats. Methods ADSCs were cultured and proliferated in vitro, then identified with immunofluorescent staining by CD44, CD114, CD34. ADSCs had been marked with BrdU for 3 d. After the rats underwent the middle cerebral artery occlusion （MCAO） for 3 d, ADSCs were stereotaxically transplanted into the lateral ventricle. Neurological Severity Scores （NSS） were evaluated before and 1, 7, 14, 21, 28 d after transplanted. Rats were euthanized at 28 d after transplanted. Brain sections were stained in TTC for surveying the infarct volume. Other sections were stained by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） method for apoptosis detection in situ. Double-staining immunohistochemistry of brain section was used to identify the cell-specific brain-derived neurotrophic factor （BDNF） expression in BrdU-reactive ADSCs. Results The majority of ADSCs expressed CD44 （mesenchymal cell marker） and were negative for CD114 （mononuclear macrophage marker）, CD34 （hematopoietic stem cell marker）. NSS in ADSCs-transplanted group at day 21 and 28 of MCAO was significantly lower than that in control group （P〈0.05）. The infarct volume showed no significant change （P〉0.05）. Double immunostaining showed that mostly grafted BrdU-reactive ADSCs had migrated to the ischemic boundary zone, survived and expressed BDNF. The number of apoptotic cells in the ADSCs-transplanted group decreased significantly compared with that of control group （P〈0.01）. Conclusion ADSCs stereotaxically transplanted into the lateral ventricle can promote the recovery of neurological function in rats with cerebral ischemia through reducing the apoptotic cells in the ischemic boundary zone.

关 键 词：脂肪组织源性基质细胞 立体定向 移植 脑缺血 

分 类 号：R651[医药卫生—外科学]