目的 探讨p53蛋白在电针预处理改善急性脑缺血再灌注期大鼠神经功能中的作用。方法 将72只Sprague-Dawley大鼠随机分成假手术组、模型组和电针预处理组,每组24只,每组再分为再灌注2 h、72 h亚组,每亚组12只。建立模型后对再灌注后2 h、72 h大鼠行神经行为学评估,HE与TUNEL染色观察缺血区病理损伤及细胞凋亡情况,Western blotting检测缺血区p-p53 (ser392)、p53、微管相关蛋白轻链3(LC3)Ⅱ的表达。结果 与模型组相比,再灌注2 h、72 h,电针预处理组神经功能缺损评分均降低(P<0.05),缺血区病理损伤程度下降,缺血半暗区皮层细胞凋亡数在再灌注72 h减少(P<0.05),缺血区p-p53、LC3 Ⅱ蛋白水平降低(P<0.05)。再灌注2 h,电针预处理组p53蛋白水平较模型组比较无显著性差异(P>0.05);而再灌注72 h时,电针预处理组p53蛋白水平降低(P<0.05)。结论 电针预处理可以改善急性脑缺血再灌注损伤大鼠神经功能缺损,其机制可能与p53蛋白参与细胞自噬及凋亡调控有关。
Objective To investigate the effect of p53 protein on the nerve function of rats with acute cerebral ischemia-reperfusion by electro-acupuncture pretreatment.Methods A total of 72 male Sprague-Dawley rats were randomly divided into sham group (n=24), model group (n=24) and electroacupuncture pretreatment group (n=24). Each group was equally divided into two hours and 72 hours subgroups. The neurological behavior was assessed with modified Neurological Severity Score (mNSS). HE staining and TUNEL assay were used to observe the injury and cell apoptosis in ischemic brain, and Western blotting was used to detect the expression of p-p53 (ser392), p53 and microtubule associated protein 1 light chain 3 (LC3)II in ischemic area.Results Compared with the model group, the mNSS scores, the injury of brain tissue, the number of TUNEL-positive cells, and the level of p-p53 and LC3II decreased (P<0.05) in the electroacupuncture pretreatment group. Two hours after reperfusion, there was no significant difference in the level of p53 between the model group and the electroacupuncture pretreatment group (P>0.05), however, 72 hours after reperfusion, the level of p53 decreased in the electroacupuncture pretreatment group (P<0.05).Conclusion Electroacupuncture pretreatment can improve neurological deficit in cerebral ischemia-reperfusion injury rats, which may be related to the involvement of p53 protein in autophagy and apoptosis
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