摘要
目的研究重组人促红细胞生成素（rhEPO）对慢性脑缺血大鼠学习记忆能力及凋亡相关蛋白P53和Bcl-2表达的影响。方法16只健康雄性SD大鼠结扎双侧颈总动脉（2VO）建立永久性慢性脑缺血模型，将其按照随机数字表法分为对照组和实验组，每组各8只。模型建立3d后，实验组每7天经鼻腔给予150U／125μLrhEP0，至建模后8周末；对照组于同一时间鼻腔给予等量的生理盐水。8周后应用Morris水迷宫观测2组大鼠的运动及空间学习记忆能力。采用HE染色观察大脑皮层及海马CAl区神经元形态学的变化，并采用图像分析软件对大脑皮层厚度与神经元数目进行比较。免疫组化法检测P53及Bcl-2蛋白的表达水平。原位末端标记（TUNEL）法检测凋亡细胞数。结果（1）行为学测试结果显示：实验组平均逃避潜伏期及穿越平台次数均较对照组有所改善，差异有统计学意义（P〈0．05）。（2）HE染色结果显示：与实验组相比，对照组大鼠皮层及海马CAl区锥体神经元胞体萎缩、核固缩且皮层厚度变薄，数目减少，差异有统计学意义（P〈0．05）。f3）免疫组化结果显示：与对照组相比，实验组Bcl．2的表达明显增强，P53平均灰度值明显增加，差异有统计学意义（P〈0．05）。f4）TUNEL染色结果显示：对照组凋亡细胞数较实验组明显增加，差异有统计学意义（P〈0．05）。结论rhEPO能改善慢性脑缺血大鼠运动、记忆及空间定向能力，这种神经功能保护作用的机制可能与减少神经元凋亡相关。
Objective To study the effects of recombinant human erythropoietin （rhEPO） on learning and memory functions and expressions of apoptosis-related proteins （P53 and Bcl-2） in rats induced by chronic cerebral ischemia. Methods Sixteen healthy male SD rats were randomly divided into control group and experimental group （n=8）; the permanent bilateral occlusion of common carotid arteries in these rats was adopted to establish the chronic cerebral ischemia models; rats in the experimental group weekly received intranasal rhEPO delivery at the dose of 150 U/125 p.L after chronic ischemia for 3 days, whereas models in the control group accepted equivalent volume of saline at the same time. Eight weeks after the inducement, Morris water maze test was performed to evaluate the movement and the spatial learning and memory capabilities. Morphology changes of cerebral cortex and hippocampal CA1 nerve cells were observed by HE staining. P53 and Bcl-2 proteins levels were detected by immunohistochemistry. The number ofapoptotic cells was detected by means of TUNEL. Results Morris water maze test showed that shorter escape latency and higher frequency through platform in the experimental group were noted as compared with those in the control group （P〈0.05）. HE stainingindicated that less pyramidal cells and more serious karyopyknosis changes of cerebral cortex and hippocampal CA1 nerve cells and thinner cerebral cortex in control group were noted as compared with those in the experimental group （P〈0.05）. Immunohistochemistry indicated that the experiment group had increased Bal-2 expression and mean gray value of P53 as compared with control group （P〈0.05）. TUNEL showed that the apoptotic cells of control group were significantly increased as compared with those in the experimental group （P〈0.05）. Conclusion The rhEPO can improve the abilities of movement, memory and spatial orientation in rats induced by chronic cerebral ischemia, whose mechanism might be related to the restrain apoptosis.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2013年第6期565-569,共5页
Chinese Journal of Neuromedicine
