苦碟子注射液对急性脑缺血大鼠脑组织ROKα表达的影响

Influence of Kudiezi Injection on brain ROKα expression in rats with acute cerebral ischemia

目的研究具有解毒通络作用的苦碟子注射液对急性脑缺血大鼠脑组织Rho相关卷曲螺旋形成蛋白激酶α(ROKα)表达的影响,探讨解毒通络法的脑保护作用机制。方法采用免疫组织化学法检测大脑中动脉闭塞(MCAO)致局灶性脑缺血模型大鼠脑组织中ROKα蛋白的表达及分布,实时荧光定量PCR法检测ROKα mRNA的表达,及其在中药苦碟子注射液、西药ROK选择性抑制剂盐酸法舒地尔注射液干预后的变化。结果 ROKα在模型大鼠缺血脑组织中异常高表达(P<0.01),苦碟子注射液可明显降低缺血皮层、海马ROKα及其mRNA的表达(P<0.01),与盐酸法舒地尔注射液比较无明显差异(P>0.05)。结论具有解毒通络作用的苦碟子注射液对急性脑缺血大鼠有脑保护作用,其作用机制与降低ROKα在缺血脑组织中的表达有关。

Objective To study the influence of Kudiezi Injection, having the actions of detoxification and dredging collaterals, on the expression of Rho-associated coiled-coil containing protein kinase α （ROKα） in rats with acute cerebral ischemia, and to investigate the brain protection mechanism of the therapy of detoxification and dredging collaterals. Methods The expression and distribution of brain ROKα were detected by using immunohistochemistry in rats with focal cerebral ischemia induced by middle cerebral artery occlusion （MCAO）. The expression of ROKα-mRNA and its changes after intervened by Kudiezi Injection or Hydroxyfasudil Injection （a selective inhibitory agent of ROK） were detected by applying real-time fluorescence quantitative polymerase chain reaction （RT-PCR）. Results The expression of ROKα was high abnormally in ischemia brain of rat model （ P 〈 0. 01 ）. Kudiezi Injection decreased significantly the expressions of ROKα and ROKα-mRNA in ischemia cortex and hippocampus （P 〈0.01 ）. The comparison between Kudiezi Injection and Hydroxyfasudil Injection did not show significant difference （P 〉 0. 05 ）. Conclusion Kudiezi Injection, having the actions of detoxification and dredging collaterals, has a brain protective effect in rats with acute cerebral ischemia, and the mechanism may be related to that it decreases the expression of ROKα in ischemia brain.