丹龙醒脑片对大鼠局灶性脑缺血再灌注后海马区肿瘤坏死因子α表达和细胞凋亡影响.docVIP

丹龙醒脑片对大鼠局灶性脑缺血再灌注后海马区肿瘤坏死因子α表达和细胞凋亡的影响 作者：何倩,周小青,李花,刘旺华,刘建新 【摘要】 目的 探讨丹龙醒脑片对大鼠脑缺血再灌注后海马区神经细胞凋亡和肿瘤坏死因子α(TNF-α)表达的影响,探讨其作用机制。方法 应用线栓法制作大鼠局灶性脑缺血再灌注模型,用原位末端标记法和免疫组化法分别检测假手术组、模型组、丹龙醒脑片组和尼莫地平组的凋亡细胞数和TNF-α阳性细胞数。结果 与假手术组相比,模型组凋亡细胞数和TNF-α表达明显升高(P＜0.05,P＜0.01)；与模型组相比,丹龙醒脑片组凋亡细胞数和TNF-α表达明显降低(P＜0.05)。结论 TNF-α表达下降可能是丹龙醒脑片减少大鼠脑缺血再灌注损伤后细胞凋亡的分子机制之一。 【关键词】 脑缺血再灌注；丹龙醒脑片；细胞凋亡；肿瘤坏死因子α；大鼠 Abstract：Objective To study the effects of Danlong Xingnao Tablet on neuronal apoptosis and expression of TNF-α after focal cerebral ischemia reperfusion in rat hippocampus. Methods The model of middle cerebral arteries occlusion (MCAO) were performed with intraluminal filament blockade. By using in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotor nick end labeling (TUNEL) and immunohistochemistry staining, the number of apoptotic cells and TNF-α positive cells were detected in the sham-operated group, the ischemia reperfusion group, Danlong Xingnao Tablet group and nimodipine group after the middle cerebral arteries of SD rats occluded for 1 h and reperfused for 24 h．Results Compared with the sham operation group, the number of apoptotic cells and TNF-α positive cells of the model group increased significantly (P＜0.05, P＜0.01). Compared with the model group, DLXNT intragastric administration could reduce the number of apoptotic cells and TNF-α positive cells (P＜0.05). Conclusion The decreased expression of TNF-α may be one of the molecular mechanisms of decreased apoptotic cell after rat cerebral ischemia reperfusion injurying treated with Danlong Xingnao Tablet． Key words：cerebral ischemia reperfusion；Danlong Xingnao Tablet；apoptosis；TNF-α；rat 丹龙醒脑片是临床治疗缺血性卒中的有效复方。以往的研究证实,丹龙醒脑片具有增加脑组织局部血流量、抗神经细胞凋亡等作用[1-3]。在局灶性脑缺血再灌注后存在大量的神经细胞凋亡,尤其在缺血半暗带区及对缺血较为敏感的海马区。肿瘤坏死因子α(TNF-α)是凋亡诱导家族的重要成员之一,本实验选用大鼠局灶性脑缺血再灌注模型,观察丹龙醒脑片对脑缺血再灌注后海马区TNF-α蛋白表达的影响,探讨丹龙醒脑片抗局灶性脑缺血再灌注神经细胞凋亡、减轻病理性损伤的机制。 　　1 实验材料 　　1.1 动物 清洁级SD大鼠72只,雌雄各半,体重280～300 g,中南大 学湘雅医学院动物学部提供,合格证号：医动字第20-010号。 　　1.2 药物 丹龙醒脑片由丹参10 g、三七6 g、地龙6 g、远志