摘要
目的探讨多奈哌齐对脑梗死后室管膜下区(subventricular zone,SVZ)神经发生相关神经营养因子表达的影响。方法成年雄性昆明小鼠78只,按随机数字表法分为假手术+溶剂组(18只)、脑梗死+溶剂组(30只)、脑梗死+多奈哌齐组(30只)。线栓法建立大脑中动脉闭塞模型(middle cerebral artery occlusion,MCAO),采用尼氏染色法测脑梗死体积,改良神经功能缺损评分评估神经功能缺损程度,脑含水量测定脑水肿程度;用5-溴-2'-脱氧尿苷(5-bromodeoxyuridine,BrdU)标记增殖细胞,双肾上腺皮质激素(doublecortin,DCX)标记神经母细胞,免疫荧光观察各组小鼠SVZ中BrdU+/DCX+细胞的表达;Western blot分析各组小鼠SVZ胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)及神经生长因子(nerve growth factor,NGF)表达量的变化。结果脑梗死+多奈哌齐组脑梗死体积[(13.33±4.55)%]较脑梗死+溶剂组[(31.33±3.93)%]显著降低(t=7.34,P<0.05),神经功能缺损明显改善,且脑含水量[(71.82±10.18)%]较脑梗死+溶剂组[(85.93±7.54)%]明显减轻,均差异有统计学意义(F=13.480,P<0.05);脑梗死+溶剂组SVZ中BrdU+/DCX+细胞荧光面积[(6.16±1.79)%]较假手术+溶剂组[(2.25±1.09)%]显著增加,脑梗死+多奈哌齐组SVZ中BrdU^+/DCX^+细胞荧光面积[(16.19±2.16)%]较脑梗死+溶剂组显著增加,均差异具有统计学意义(F=102.756,P<0.05);脑梗死+多奈哌齐组小鼠SVZ中GDNF、BDNF及NGF的表达量较脑梗死+溶剂组显著增多,脑梗死+溶剂组上述蛋白表达量较假手术+溶剂组显著增加,均差异有统计学意义(F=15.114,27.121,27.398,均P<0.05)。结论多奈哌齐通过促进脑梗死后SVZ中GDNF、BDNF及NGF等神经营养因子的表达调控神经发生。
Objective To investigate the effect of donepezil on subventricular zone(SVZ)neurogenesis related neurotrophic factors after cerebral infarction.Methods Mice were randomly assigned into three groups: vehicle-treated sham group(Sham+vehicle,n=18),vehicle-treated middle cerebral artery occlusion(MCAO)group(MCAO + vehicle,n=30)and donepezil-treated MCAO group(MCAO + donepezil,n=30).Middle cerebral artery occlusion(MCAO)was induced by thread-occlusion method.Nissl staining was used to measure the infarct volume and the modified neurological severity score(mNSS)was used to assess neurologic function and brain water content was detected to assess brain edema degree.Proliferative cells and neuroblasts were labeled with 5-bromodeoxyuridine(BrdU)and doublecortin(DCX).The SVZ BrdU^+ /DCX^+ cells were detected by immunofluorescence.The expression of glial cell line-derived neurotrophic factor(GDNF),brain-derived neurotrophic factor(BDNF)and nerve growth factor(NGF)were detected by Western blot.Results The infarct volume of MCAO + donepezil group((13.33±4.55)%)was significantly lower than that of MCAO + vehicle group((31.33±3.93)%,t=7.34,P<0.05).The neurologic deficits were significantly ameliorated after donepezil treatment,and the brain water content of MCAO + donepezil group((71.82±10.18)%)was significantly less than that of MCAO + vehicle group((85.93±7.54)%,F=13.480,P<0.05).All differences were statistically significant(P<0.05).The area of BrdU^+ /DCX^+ cells within SVZ of MCAO + vehicle group((6.16±1.79)%)was significantly larger than that of sham + vehicle group((2.25±1.09)%),and was fewer than that of MCAO^+ donepezil group((16.19±2.16)%,F=102.756,P<0.05).MCAO significantly promoted the expression of GDNF,BDNF and NGF within SVZ compared with sham operation,and donepezil increased these protein levels(F=15.114,27.121,27.398,P<0.05).Conclusion Donepezil regulates neurogenesis via increasesing the expression of GDNF,BDNF and NGF within SVZ after cerebral infarction.
作者
张永鑫
王建平
崔可飞
付晓杰
张帝
鲁争芳
高宇峰
满江
刘贤良
李思佳
廖羚慧
李想
Zhang Yongxin;Wang Jianping;Cui Kefei;Fu Xiaojie;Zhang Di;Lu Zhengfang;Gao Yufeng;Man Jiang;Liu Xianliang;Li Sijia;Liao Linghui;Li Xiang(Department of Neurology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Ultrasound,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Nerve Intervention,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2018年第11期961-965,共5页
Chinese Journal of Behavioral Medicine and Brain Science
基金
国家自然科学基金项目(81571137,81771247)
2016~2020卫生计生科技创新型人才“51282”工程
河南省医学科技攻关计划项目(201501005).
关键词
多奈哌齐
脑梗死
室管膜下区
神经发生
神经营养因子
Donepezil
Cerebral infarction
Subventricular zone
Neurogenesis
Neurotrophic factors
