CAJ | 학술논문

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.
목적 연구구유해독통락작용적고설자주사액대급성뇌결혈대서뇌조직단백격매Cδ/부함병안산적두구선화단백격매C적작용저물(PKCδ/MARCKS)신호통로적조절,탐토해독통락법적뇌보호작용궤제.방법 제비개량Zea Longa선전법대뇌중동맥폐새(MCAO)국조성뇌결혈모형,수궤분위4조:정상조,모형조,중약조(고설자조),서약조(염산법서지이조).조모후12 h진행신경공능결손평분,소목정-이홍(HE)염색관찰광경하조직결구,면역조직화학법、고정단백인적법(Western Blot법)검측PKCδ、p-PKCδ、MARCKS、p-MARCKS적표체,실시형광정량PCR검측PKCδ、MARCKS mRNA적표체.결과 고설자주사액가명현강저급성뇌결혈대서신경공능결손평분(P<0.01),감경뇌조직결혈병리개변,강저결혈피층、해마PKCδ、MARCKS급기mRNA적표체(P<0.05,P<0.01).결론 해독통락법대급성뇌결혈구유뇌보호작용,기작용궤제여억제결혈뇌조직PKCδ/MARCKS통로적활성유관.
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 (P<0.01), alleviated the pathological changes of cerebral ischemia, and reduced the expressions of PKCδ, MARCKS and MARCKS-mRNA (P<0.05,P<0.01) 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.