脐血源神经干细胞在SDF-1/CXCR4趋化下对脑出血大鼠神经损伤恢复的作用  

Effects of umbilical cord blood neural stem cells via stromal cell-derived factor-1/CXC chemokine receptor 4 signaling on neural recovery in rat models of intracerebral hemorrhage

作　　者：马建华 李鑫 戴宜武 Ma Jianhua;Li Xin;Dai Yiwu(Department of Vascular Surgery,Feicheng People's Hospital of Shandong Province,Feicheng 271600,China;Department of Neurosurgery,Seventh Medical Center,General Hospital of People's Liberation Army,Beijing 100700,China)

机构地区：[1]山东省肥城市人民医院血管外科,271600 [2]解放军总医院第七医学中心神经外科,北京100700

出　　处：《中华神经医学杂志》2019年第4期344-350,共7页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金(81271391).

摘　　要：目的探讨移植的脐血源神经干细胞(UCBNSCs)在基质细胞衍生因子-1(SDF-1)/CXC趋化因子受体4(CXCR4)作用下对脑出血大鼠神经功能修复的影响。方法(1)体外迁移实验中,将UCBNSCs置入Transwell小室的上室,在下室中分别加入0、30、60和120ng/mL浓度的SDF-1。(2)体内实验中,将60只脑出血模型大鼠按照随机数字表法分为UCBNSCs移植组和对照组,每组30只。造模后2d分别向2组大鼠颅内移植UCBNSCs悬液10μL和等量的磷酸盐缓冲液(PBS),同时腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)标记内源性神经干细胞。分别于移植后1d、1周、2周对2组大鼠进行改良神经功能缺损评分(mNSS)。移植后2周处死全部大鼠,进行SDF-1、BrdU、血管内皮生长因子(VEGF)、神经胶质纤维酸性蛋白(GFAP)及双肾上腺皮质激素(DCX)免疫荧光染色;同时采用转移酶介导的三磷酸脱氧鸟苷-生物素刻痕末端标记(TUNEL)检测试剂盒检验细胞凋亡情况。结果(1)体外实验中,UCBNSCs中能够检测到CXCR4的表达;60ng/mLSDF-1对UCBNSCs迁移作用最大,与其他浓度SDF-1比较,差异有统计学意义(P<0.05)。(2)体外实验中,移植2周后,UCBNSCs组大鼠室管膜下区BrdU标记的细胞数量增多,BrdU/DCX、BrdU/GFAP细胞数量明显比对照组增多,差异有统计学意义(P<0.05);UCBNSCs移植组脑损伤区神经营养因子VEGF阳性细胞数[(88.30±7.21)个/视野]明显高于对照组[(53.20±4.45)个/视野],差异有统计学意义(t=4.144,P=0.000);UCBNSCs移植组脑损伤区凋亡细胞的数量[(34.30±2.44)个/视野]明显低于对照组[(47.70±1.98)个/视野],差异有统计学意义(t=4.266,P=0.001)。移植后2周时UCBNSCs组mNSS评分[(6.40±0.163)分]明显低于对照组[(7.50±0.17)分],差异有统计学意义(t=4.714,P=0.002)。结论SDF-1/CXCR4可以趋化移植的UCBNSCs到达脑出血损伤区,促进大鼠神经功能恢复,其作用机制可能与促进神经发生、VEGF的分泌和抑制细胞凋亡有关。Objective To investigate the effects of umbilical cord blood neural stem cells(UCBNSCs)via stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)signaling on neural recovery in rat models of intracerebral hemorrhage.Methods(1)In migration assay in vitro,UCBNSCs were distributed in the upper wells of Transwell plates,and SDF-1 at concentrations of 30,60 and 120 ng/mL was placed in the lower wells.(2)Sixty rat models of intracerebral hemorrhage were randomly divided into UCBNSCs-transplanted group and phosphate buffer(PBS)-transplanted group(n=30);two d after modeling,10 L UCBNSCs suspension and same amount of PBS were,respectively,transplanted into the two groups,and intraperitoneal injection of deoxyuridine(BrdU)labeled endogenous neural stem cells was performed;neurological functions were assessed with modified Neurological Severity Scale(mNSS)one d,and one and two weeks after cell transplantation;the expressions of SDF-1,vascular endothelial growth factor(VEGF),glial fibrillar acidic protein(GFAP),and doublecortin(DCX)were detected by immunofluorescence when the rats were sacrificed two weeks after cell transplantation;cell apoptosis was detected by TUNEL.Results(1)In the in vitro experiment,CXCR4 expression could be detected in UCBNSCs;60 ng/mL SDF-1 had the greatest migration effect on UCBNSCs,and this effect showed statistically significant difference as compared with that at other concentrations(P<0.05).(2)In the in vivo experiment,two weeks after transplantation,the UCBNSCs-transplanted group had significantly increased number of BrdU-labeled cells in the subventricular zone,and significantly larger number of BrdU/DCX and BrdU/GFAP cells than the PBS-transplanted group(P<0.05);the VEGF expression in the brain injury area of the UCBNSCs-transplanted group([88.30±7.21]/field)was significantly higher than that of PBS-transplanted group([53.20±4.45]/field,t=4.144,P=0.000);the number of apoptotic cells in the brain injury area of the UCBNSCs-transplanted group([34.30±2.44]/field)was significant

关 键 词：脐血源神经干细胞 基质细胞衍生因子-1/CXC趋化因子受体4 脑出血 神经损伤 

分 类 号：R743.34[医药卫生—神经病学与精神病学]