9601颗粒对急性多发脑梗塞大鼠海马皮层MARCKS磷酸化调节作用的研究

目的研究急性多发脑梗塞大鼠海马皮层MARCKS及p-MARCKS蛋白的表达,进一步观察9601颗粒对MARCKS信号转导通路的影响,从蛋白水平探讨中药对急性脑缺血保护作用机制.方法采用改良Kaneko法建立急性多发脑梗塞动物模型,观察造模后各组大鼠的神经系统症状体征积分改变及海马皮层组织的病理变化及细胞超微结构的改变;应用免疫组织化学和固定化蛋白印迹法检测急性缺血后海马皮层MARCKS及p-MARCKS蛋白表达的改变以及中药的调节作用.结果 MARCKS及p-MARCKS蛋白在急性缺血后海马皮层中表达比正常对照组明显升高(P＜0.05),应用9601和尼莫地平可以使海马皮层异常升高的MARCKS及p-MARCKS表达下降,但中西药之间无显著差异(P＞0.05).结论在急性脑缺血状态下大鼠海马皮层MARCKS和p-MARCKS蛋白表达异常升高,与缺血损害关系密切,9601颗粒对此具有明显的下调作用.

Study on regulating effects of 9601-Granules on phosphorization of MARCKS in hippocampal cortex in the rats with multiple acute cerebral infarct

Objective To study the expressions of MARCKS and p-MARCKS in the hippocampal cortex in the rats with multiple acute cerebral infarct, observe the effects of 9601-Granules on MARCKS signal transduction pathway, and investigate the protective mechanism of TCM drugs against acute cerebral ischemia on the basis of proteins. Methods The rat model of multiple acute cerebral infarct was established by using modified Kaneko method; changes in the scoring of symptoms and signs of nervous system, and changes in the pathology and cell ultrastructure of hippocampal cortex, were observed in each experimental group; changes in the expressions of MARCKS and p-MARCKS in hippocampal cortex experienced acute ischemia and the regulating effects of TCM drugs on their expressions were detected by using immunohistochemical method and Western blot method. Results The levels of the expressions of MARCKS and p-MARCKS in hippocampal cortex were higher in the group experienced acute ischemia than those in the normal control group (P<0.05). Although both 9601-Granules and Nimodipine could decrease the abnormal elevation in the expressions of MARCKS and p-MARCKS, there was no significant difference between the effects of the two drugs. Conclusion Under the condition of acute cerebral ischemia, the expressions of MARCKS and p-MARCKS are abnormally increased in hippocampal cortex in the rats, which is closely related with ischemic defect; and 9601-Granules possess marked downregulating effects on the abnormal increase.