解毒通络法对急性脑缺血大鼠脑组织PKCδ/MARCKS信号通路的调节  

Regulation of therapy of dispelling toxin and dredging collaterals on PKCδ/MARCKS signal pathway in rats with acute cerebral ischemia

作　　者：白文[1] 杨嘉颐[1] 张允岭[2] 王今芳[1] 柳洪胜[1] 戴中[1] 王春玲[1] 郑宏[2] 王少杰[1] 

机构地区：[1]北京大学人民医院,北京100044 [2]北京中医药大学东方医院

出　　处：《北京中医药大学学报》2010年第2期88-93,F0003,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家重点基础研究发展计划(973计划)资助项目(No.2006CB504805);北京大学人民医院研究与发展基金资助项目(No.RDB2007-50)

摘　　要：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蛋白激酶Cδ/富含丙氨酸的豆蔻酰化蛋白激酶C的作用底物(PKCδ/MARCKS)信号通路的调节,探讨解毒通络法的脑保护作用机制。方法制备改良Zea Longa线栓法大脑中动脉闭塞(MCAO)局灶性脑缺血模型,随机分为4组:正常组,模型组,中药组(苦碟子组),西药组(盐酸法舒地尔组)。造模后12 h进行神经功能缺损评分,苏木精-伊红(HE)染色观察光镜下组织结构,免疫组织化学法、固定蛋白印迹法(Western Blot法)检测PKCδ、p-PKCδ、MARCKS、p-MARCKS的表达,实时荧光定量PCR检测PKCδ、MARCKS mRNA的表达。结果苦碟子注射液可明显降低急性脑缺血大鼠神经功能缺损评分(P<0.01),减轻脑组织缺血病理改变,降低缺血皮层、海马PKCδ、MARCKS及其mRNA的表达(P<0.05,P<0.01)。结论解毒通络法对急性脑缺血具有脑保护作用,其作用机制与抑制缺血脑组织PKCδ/MARCKS通路的活性有关。Objective To study the regulation of Kudiezi Injection with the actions of dispelling toxin and dredging collaterals on PKCδ/MARCKS signal pathway in rats with acute cerebral ischemia,and investigate the brain protective mechanism of the therapy of dispelling toxin and dredging collaterals.Methods The rat model of local cerebral ischemia induced by middle cerebral artery occlusion（MCAO） was established by using modified Zea Longa suture.All rats were randomly divided into the control group,model group,Chinese medicine group（with Kudiezi Injection） and Western medicine group（with hydroxyfasudil injection）.The neurological functional deficit score was reviewed in all groups 12 hours after model establishment.The pathology of cerebral tissue was observed by the light microscope after HE staining.The expressions of PKCδ,p-PKCδ,MARCKS and p-MARCKS were detected by using immunohistochemistry and Western blotting,and the expressions of PKCδ and MARCKS-mRNA were detected by using real-time fluorescence quantitative polymerase chain reaction（RT-PCR）.Results Kudiezi Injection decreased significantly the neurological functional deficit score（P0.01）,alleviated the pathological changes of cerebral ischemia,and reduced the expressions of PKCδ,MARCKS and MARCKS-mRNA（P0.05,P0.01） in rats with acute cerebral ischemia.Conclusion The therapy of dispelling toxin and dredging collaterals has a brain protective effect in the treatment of acute cerebral ischemia,and the mechanism may be related to that the therapy can inhibit the activity of PKCδ/MARCKS signal pathway in ischemia brain.

关 键 词：解毒通络 苦碟子注射液 局灶性脑缺血 PKCδ/MARCKS 脑保护 大鼠 

分 类 号：R285.5[医药卫生—中药学]