重组人粒细胞集落刺激因子对缺血缺氧性脑损伤新生大鼠凋亡诱导因子表达的影响

摘要

目的:研究分析重组人粒细胞集落刺激因子对新生SD大鼠凋亡诱导因子表达的影响.方法:随机选取7日龄SD大鼠36只,随机分为假手术组(A组)、脑缺血再灌注组(B组)和药物治疗组(C组),每组各12只.采用Longa线栓法制作大鼠大脑动脉闭塞再灌注模型,C组大鼠在脑缺血2 h后及24 h时给予药物灌注,其余2组给予大鼠注射等量的0.9%氯化钠注射液.实验结束后,采用Longa 5分制标准评分法在24 h时对大鼠的神经功能进行评分,采用免疫组织化学法对大鼠的组织磷酸化c-jun氨基末端激酶、磷酸化细胞外信号调节蛋白激酶进行检测,同时采用原位末端转移酶标记对神经细胞凋亡、TTC染色测脑梗死体积进行相关检测.对凋亡相关基因的表达及凋亡水平进行评价.结果:A组仅有一小部分凋亡细胞,B、C 2组在灌注24 h后可以发现大量的凋亡细胞,细胞核均呈棕褐色,主要分布在缺血脑组织中周围;C组脑梗死程度较B组减轻(P<0.01).结论:重组人粒细胞集落刺激因子可以起到保护新生SD大鼠神经细胞的作用,避免发生缺血缺氧性脑损伤,这种糖蛋白与细胞凋亡相关基因的表达有关.%Objective:To study the effects of recombinant human granulocyte colony stimulating factor( rhG-CSF) on the expression of apoptosis inducing factor( AIF) in neonatal SD rats. Methods:Thirty-six healthy neonatal SD rats with 7 days old were randomly divided into the sham operation group(group A),cerebral ischemia reperfusion group(group B) and drug treatment group(group C) (12 rats each group). The rat cerebral arterial occlusion reperfusion model was made using Longa suture blot method. The group C were injected with drug after two hours and 24 hours of cerebral ischemia,and the other two groups were injected with the same amount of normal saline. At the end of the experiment, the neural function of rat was evaluated by Longa 5 score method, the expressions of extracellular signal regulated protein kinase(p-ERK) and phosphorylated extracellular signal regulated protein kinase(p-JNK) were detected by immunohistochemical method,and the neuronal apoptosis and cerebral infarction volume were detected using Tunel and TTC staining,respectively. The levels of expression and apoptosis of apoptosis related genes were evaluated. Results:A small part of the apoptotic cells in A group and large number of apoptotic cells and brown nuclei in groups B and C after 24 h of reperfusion were found, which distributed mainly around the ischemic brain tissue,and which in C group was less than that in group B(P < 0. 01). Conclusions:rhG-CSF can protect the neuron of neonatal rat, and avoid the occurrence of ischemic hypoxic brain damage, which is related to the expression of the apoptosis related gene.