南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (05): 564-.

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成年大鼠全脑缺血性脑损伤后神经前体细胞的增殖周期变化

李鸥; 朱心红; 高天明;   

  1. 南方医科大学神经生物学教研室; 南方医科大学神经生物学教研室 广东广州510515; 广东广州510515;
  • 出版日期:2006-05-20 发布日期:2006-05-20
  • 基金资助:
    国家自然科学基金;广东省自然科学基金;教育部“长江学者”奖励计划~~

Periodic quantity variation of proliferating neuronal progenitors in adult rats after global brain ischemia

LI Ou,ZHU Xin-hong,GAO Tian-ming Department of Neurobiology,Southern Medical University,Guangzhou 510515,China   

  1. 南方医科大学神经生物学教研室; 南方医科大学神经生物学教研室 广东广州510515; 广东广州510515;
  • Online:2006-05-20 Published:2006-05-20

摘要: 目的研究大鼠全脑缺血后内源性神经前体细胞增殖周期的变化规律,为确定促脑神经元再生药物干预时间窗的选择提供依据。方法采用改良的Pulsinelli四血管闭塞法制作前脑缺血模型,成年雄性Wistar大鼠随机分为8组(n=3)。分别于缺血后第7~14天,单日4次间隔2h腹腔注射BrdU75mg/kg。缺血第29天灌注固定,另取3只正常大鼠以同样方法注射BrdU,次日处死。取脑,连续冰冻冠状切片,分别做BrdU免疫组化标记及BrdU/NeuN荧光免疫组化双标记并计数BrdU+细胞密度及BrdU+/NeuN+细胞密度。采用单因素方差分析,比较缺血后不同时间增殖细胞数量的差异。结果缺血性脑损伤促进了大鼠神经前体细胞的分裂,海马齿状回DG区和CA1区的新生细胞在缺血后第7~10天处于较高的水平且逐渐减少,11d后降至稳定水平。神经前体细胞的分化未受缺血后时间变化的影响。结论全脑缺血对神经前体细胞的促分裂作用主要发生在缺血后10d内。 

Abstract: Objective To investigate the periodic quantity variation of proliferating neuronal progenitors after global brain ischemia and provide evidence for choosing the time-window of drug therapy to promote neuronal regeneration after ischemia.Methods Adult male Wistar rats were subjected to 15-min global brain ischemia(four-vessel occlusion model)and randomized subsequently into 8 groups(n=3).The rats were given intraperitoneal injections of BrdU(75 mg/kg)for 4 times daily(at a 2-hour interval)since day 7 till day 11 after ischemia,and on day 29,the rats were perfused transcardially for fixation.Another 3 normal rats were given BrdU in the same manner and killed the next day.Coronal sections of the brain tissue(30 μm)were prepared for immunocytochemical detection of BrdU-labeled cells and immunofluorescent detection of BrdU/NeuN double-labeled cells.The density of BrdU-positive cells and BrdU/NeuN double-labeled cells in the hippocampal dentate gyrus(DG)and CA1 region were counted and the density of proliferating cells at different days after ischemia were compared using one-way ANOVA.Results The proliferation of the neuronal progenitors increased after global brain ischemia.The number of BrdU-positive cells in the DG and CA1 region decreased gradually in 7-10 days after ischemia,and reached the normal level during 11-14 days.The differentiation of the progenitors did not vary after ischemia.Conclusion Increased proliferation of the neuroprogenitors occurs mainly within the initial 10 days after global ischemia in rats. 

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