镇肝熄风汤预处理对MCAO大鼠脑组织病理学变化和神经元凋亡的影响

Effect of Zhengan Xifeng Decoction Pretreatment on Pathological Changes and Neuron Apoptosis of Cerebral Tissue in Rat

摘要:

目的: 研究镇肝熄风汤对大脑中动脉梗死(MCAO)大鼠脑组织的病理学变化和神经元凋亡的影响。 方法: MCAO术前给予高、低剂量镇肝熄风汤(15,30 g·kg^-1)灌胃14 d,采用线栓法制备MCAO。术后6 h称重法检测脑水肿程度,TTC染色观察脑梗死范围、HE染色观察缺血组织病理形态和海马神经元密度,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)及Western bolt检测凋亡效应蛋白半胱胺酸蛋白酶蛋白(Caspase 3)和聚ADP核糖聚合酶(PARP)表达情况。 结果: 与假手术组比较,模型组大鼠出现行为障碍,梗死面积及缺血侧脑组织含水量显著增加,海马CA1区神经元密度明显降低。与模型组比较,镇肝熄风汤高、低剂量均能有效改善大鼠行为学积分,可明显改善MCAO大鼠缺血侧脑组织含水量,减少梗死面积,改善局灶性脑缺血引起的海马CA1区神经元密度降低。镇肝熄风汤预处理后,明显减少MCAO大鼠缺血侧脑组织神经元细胞凋亡率,明显降低缺血侧脑组织活化Caspase-3和PARP蛋白表达量。 结论: 镇肝熄风汤可能通过影响神经细胞凋亡,发挥保护缺血大鼠脑组织的作用。

Abstract:

Objective: To study the effect of Zhengan Xifeng decoction (ZGXFD) on pathological changes and neuron apoptosis in cerebral tissuein of middle cerebral artery occlusion(MACO) in rats. The effect of ZGXFD on the behavior obstade in rats was observed. Method: The model of focal cerebral ischemia in rats was established with the suture-occuluded method and was used to intervene with different dose of ZGXFD (15,30 g·kg^-1,respectively)before MCAO. The effect of ZGXFD on the behavior obstacle in rats was observed, the infarct size and the pathological change were observed by TTC staining, and HE dyeing,respectively. TUNEL was used to evaluate the apoptosis of neurons, and Western blot was used to observe the expression of Caspase 3 and poly ADP-ribose polymerase(PARP). Result: After modeling, behavior obstacle with different degree appeared in all rats. The infarct size could be observed in all groups except sham group. The nerve cell apoptosis could be found in all rats after 6 h of MCAO. The result showed that the ZGXFT improved the behavior obstacle, reduced the infarct size and cerebral edema obviously;and the AI and apoptosis of Caspase-3 and PARP were decreased significantly after 6 h MCAO. Conclusion: ZGXFD pretreatment has obvious protective effects on the focal cerebral ischemia in rats.