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目的:氟桂利嗪是通过血脑屏障的细胞钙超载阻滞剂。探其对迟发性神经元死亡发生后沙土鼠空间分辨学习的影响及对脑海马CA1区神经元的保护作用。方法:实验于2004-03/06在大连医科大学基础医学院生理实验室完成。将健康雄性蒙古沙土鼠56只随机分3组:假手术组(n=8),缺血再灌注组(n=24),氟桂利嗪组(n=24),后两组又分为缺血5min后再灌注2d及7d组和Y迷宫组3个亚组,每亚组8只动物。缺血再灌注组和氟桂利嗪组夹闭双侧颈总动脉制备缺血再灌注模型,假手术仅暴露双侧颈总动脉,不夹闭。氟桂利嗪组术后胃灌氟桂利嗪5mg/kg,1次/d,直至处死前1d;其他两组胃灌生理盐水0.5mL。术后第4天利用Y迷宫检测动物空间分辨学习能力,包括学习获得所需天数及条件反射次数。实验沙土鼠脑组织结构变化及细胞内显微结构变化以免疫组织化学染色法及电镜技术桧测观察。结果:经补充56只动物进入结果分析。①Y迷宫学习获得所需天数:缺血再灌注组多于假手术组和氟桂利嗪组(5.88±0.99,2.00±0.76,2.13±0.64,P<0.01),假手术组和氟桂利嗪组比较无差异(P>0.05)。②Y迷宫学习获得条件反射次数:3组间无差异(P>0.05)。③术后7d脑海马CA1区存活神经元数:缺血再灌注组少于假手术组和氟桂利嗪组犤(8.95±2.29),(65.04±8.09),(42.13±5.90)个/视野,P<0.01犦,氟桂利嗪组仍少于假手术组(q=10.94,P<0.01)。④脑组织病理变化:电镜观察可见迟发性神经元死亡早期形成大量的多泡体,并广泛分布于CA1区神经元树突,是凋亡早期的特征性表现,5mg/kg氟桂利嗪可明显减少CA1区神经元的脱失。结论:脑缺血5min造成海马CA1区神经元选择性迟发性死亡,同时产生空间分辨学习能力的可逆性损害。5mg/kg的氟桂利嗪治疗性用药在一定程度上可以阻止迟发性神经元死亡,同时对学习、记忆等高级神经功能具有保护性作用。
Purposive: Flunarizine is a calcium overload blocker that crosses the blood-brain barrier. Explore its impact on the spatial differentiation of gerbils after delayed neuronal death and its protective effect on the hippocampal CA1 neurons. Methods: The experiment was performed at the Physiology Laboratory, School of Basic Medicine, Dalian Medical University from March to June 2004. Fifty-six healthy Mongolian gerbils were randomly divided into three groups: sham operation group (n = 8), ischemia reperfusion group (n = 24) and flunarizine group (n = 24) After ischemia for 5min, rats were reperfused with 3 subgroups of 2d and 7d and Y-maze, 8 animals in each subgroup. Ischemia / reperfusion group and flunarizine group were occluded bilateral common carotid artery ischemia-reperfusion model, bilateral surgery only exposed carotid artery, not clamping. Flunarizine group postoperative stomach irrigation Flunarizine 5mg / kg, 1 / d until 1d before death; the other two groups gastric perfusion saline 0.5mL. On the 4th postoperative day, the Y maze was used to detect the spatial learning ability of animals, including the number of days required for learning and the number of conditioned reflexes. Changes of brain tissue structure and intracellular microarchitecture in experimental gerbils were observed by immunohistochemical staining and electron microscopy. Results: 56 animals were included in the result analysis. (1) The number of days required for learning the Y-maze was higher in the ischemia-reperfusion group than in the sham-operated and flunarizine groups (5.88 ± 0.99, 2.00 ± 0.76, 2.13 ± 0.64, P <0.01) There was no difference between the two groups (P> 0.05). ②Y maze learning to obtain conditioned reflex times: no difference between the three groups (P> 0.05). (3) The number of neurons in hippocampal CA1 area at 7th day after operation was less than that in sham group and flunarizine group (8.95 ± 2.29, 65.04 ± 8.09, 42.13 ± 5.90) / field , P <0.01 犦, flunarizine group is still less than the sham group (q = 10.94, P <0.01). ④ Pathological changes of brain tissue: Electron microscopy showed that a large number of multifollicular bodies were formed in the early stage of delayed neuronal death and were widely distributed in the dendrites of neurons in CA1 area. It is a characteristic manifestation of apoptosis in the early stage. Flunarizine 5 mg / kg Can significantly reduce the loss of neurons in the CA1 area. CONCLUSIONS: Cerebral ischemia 5 min caused selective late death of neurons in hippocampal CA1 region, and at the same time resulted in reversible damage of spatial discrimination learning ability. Flunarizine 5mg / kg therapeutic drug to a certain extent, can prevent delayed neuronal death, while learning and memory, and other senior neurological protective effect.