葛根素对大鼠脑复苏后海马CA1区神经细胞凋亡相关基因的影响

背景:Fas及 P53是重要的促进凋亡的调控基因,属于肿瘤坏死因子 /神经生长因子受体家族,其表达产物具有传递凋亡信号的作用,可调控脑缺血再灌注损伤时细胞的凋亡,而葛根素则能够减轻细胞的凋亡程度。目的:观察葛根素对大鼠脑复苏后海马 CA1区神经细胞凋亡相关基因Fas及 P53的影响。设计:随机对照实验。单位:华中科技大学同济医学院附属同济医院急诊科。材料:实验于 2001-09/2002-02在华中科技大学同济医学院附属同济医院急诊科实验室完成。选取清洁级 3个月龄 W istar大鼠 45只,随机分为假手术组、模型对照组、葛根素治疗组,15只 /组。方法:葛根素治疗组及模型对照组建立大鼠急性全脑缺血再灌注模型。假手术组只分离第一颈椎两侧翼小孔,但不电凝两侧椎动脉,只分离两侧颈总动脉,但不夹闭之。葛根素治疗组于缺血前 1h 给予葛根素注射液 100m g/kg,模型对照组给予等量生理盐水,假手术组不给药。各组大鼠分别于脑缺血再灌注后 3,6,12,24,48h 即速处死,每次每组 3只。分离海马组织,制备组织切片,利用免疫组化法、原位末端标记法检测各组大鼠脑缺血再灌注后不同时间点 Fas及 P53蛋白表达的水平及凋亡细胞数的变化。主要观察指标:①各组脑缺血再灌注后不同时间点海马 CA1区 Fas及P53蛋白表达的阳性细胞数。②各组脑缺血再灌注后不同时间点海马CA1区凋亡细胞数的比较。结果:实验纳入大鼠 45只,全部进入结果分析。①各组脑缺血再灌注后不同时间点海马 CA1区 Fas蛋白表达的阳性细胞数:假手术组未见明显Fas 基因表达。与模型对照组比较,葛根素治疗组于脑缺血再灌注后各时间点均明显降低,6,12,24,48h 时差异显著 (15.0±4.3),(13.5±4.9);(40.7±3.4),(27.2±3.1);(37.0±4.8),(22.0±2.1);(24.7±4.1),(18.9±5.3)个 /m m ;P <0.05,P <0.01]。②各组脑缺血再灌注后不同时间点海马 CA1区P53蛋白表达的阳性细胞数:假手术组未见明显 P53基因表达。与模型对照组比较,葛根素治疗组于脑缺血再灌注后 24,48h 均明显降低(25.3±4.4),(12.8±2.7);(24.3±3.6),(10.9±3.0)个 /m m ;P <0.01]。③各组脑缺血再灌注后不同时间点海马 CA1区凋亡细胞数的比较:与模型对照组比较,葛根素治疗组于脑缺血再灌注后 12,24,48h 均明显降低(34.0±3.7),(21.0±3.7);(41.0±4.2),(33.0±4.8);(71.0±5.5),(41.0±3.4)个 /m m ;P <0.01]。结论:给予葛根素治疗的大鼠缺血再灌注 6～48h 时 Fas的表达显著降低,缺血再灌注 24～48h 时 P53的表达亦明显减少,且凋亡细胞数也呈下降趋势,进一步证实葛根素的脑保护作用,提示葛根素抑制脑复苏后的细胞凋亡可能与其减少促凋亡基因 Fas及 P53蛋白表达有关。

Effect of puerarin on apoptosis-related gene of nerve cell in hippocampus CA1 region of rats after cerebral resuscitation

BACKGROUND: Being a traditional Chinese herb, huangqi (astragalus membranaceus) resists free radical, enhances immunity, promotes microcirculation and protects vascular endothelial cell and nervous system; of which, the protection of huangqi (astragalus membranaceus) on nervous vessel is related to its regulation on local cerebral cortical blood flow rate. OBJECTIVE: To observe the effects of huangqi (astragalus membranaceus) on local cerebral cortical blood flow rate after traumatic injury. DESIGN: Randomized paired experiment.SETTING: Neurological Surgical Institute of Lanzhou Military Area Command of Chinese PLA.MATERIALS: The experiment was performed in Traumatic Experimental Room of Neurological Surgical Institute of Lanzhou Military Area Command of Chinese PLA from February to May 2000, in which, 75 healthy male SD rats were employed and randomized into the control (5 rats), saline control (35 rats) and experimental group (35 rats). The latter two groups were subdivided into 7 time spots of 1, 3, 6, 8, 12, 24 and 48 hours after traumatic injury, 5 rats were involved in each spot.METHODS: In normal control, the model was not prepared and in the other two groups, the modified Feeney's method was used to establish craniocerebral traumatic model. In experimental group, right after injury, huangqi injection (5 g/kg) was applied abdominally and in saline control, physiological saline 0.5 mL was injected abdominally. MAIN OUTCOME MEASURES: It was to determine the blood flow rates in local cerebral cortex at corresponding time spots after traumatic injury. Fenestration was done in normal control for direct determination. RESULTS: By supplemented, 75 rats entered result analysis. Blood flow rates in local cerebral cortex: the rate in saline control 1 hour after trauma was lower than the control (6.90±0.68), (7.94±0.65), P < 0.05], it was de creased to the minimum in 24 hours and began increasing in 48 hours (5.86±0.61), (6.15±0.60)]; the rate in experimental group at every time spot was higher than saline control (P < 0.05). CONCLUSION: Huangqi (astragalus membranaceus) increases remark ably blood flow rate in local cerebral cortex, which is associated with its neurovascular protection.