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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。
Objective To study the expression of MARCKS and p-MARCKS protein in hippocampal cortex of rats with acute multiple cerebral infarction, and further observe the effect of 9601 particles on MARCKS signal transduction pathway. To explore the protective mechanism of traditional Chinese medicine on acute cerebral ischemia from protein level. Methods The animal model of acute multiple cerebral infarction was established by the modified Kaneko method. The changes of neurological symptoms and signs and the changes of pathological changes and ultrastructure of the hippocampus in the hippocampal cortex were observed after immunization. The immunohistochemistry and immobilization were used. Western blotting was used to detect the changes of MARCKS and p-MARCKS protein expression in hippocampus after acute ischemia and the regulating effect of traditional Chinese medicine. Results The expression of MARCKS and p-MARCKS protein in hippocampus after acute ischemia was significantly higher than that in normal control group (P<0.05). MARCKS and p-MARCKS in abnormal hippocampal cortex could be increased by application of 9601 and nimodipine. The expression decreased, but there was no significant difference between Chinese and Western medicines (P>0.05). Conclusion The expression of MARCKS and p-MARCKS protein in rat hippocampus cortex is increased in acute cerebral ischemia, and it is closely related to ischemic damage. The 9601 particles have a significant down-regulation effect.