化痰通络法对急性脑梗死大鼠rt-PA溶栓后MMP-9酶原活化途径中金属蛋白酶MMP3与TIMP-1基因与蛋白表达的影响  

Effect of Huatan Tongluo on Expression of MMP-3 and TIMP-1 on Zymogen Activation pathway of MMP-9 in Rat Cortex Accepted Thrombolysis therapy after the Acute Cerebral Infarction

作　　者：周震[1] 张玉莲[1] 韩文文[1] 张琳琳[1] 宋宛珊[2] 王凯[2] 

机构地区：[1]天津中医药大学第二附属医院,天津300150 [2]天津中医药大学,天津300193

出　　处：《时珍国医国药》2014年第11期2802-2805,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81072796);天津自然科学基金(No.12JZDJC25300)

摘　　要：目的观察化痰通络法对急性脑梗死大鼠rt-PA溶栓后MMP-9酶原活化途径中金属蛋白酶MMP3与TIMP-1基因与蛋白表达的影响,为化痰通络法防治急性脑梗死溶栓后出血转化提供实验依据。方法选择健康雄性SD大鼠240只,随机分为6组,即假手术组、模型组、rt-PA组、化痰通络法(分高、常规、低中药剂量)联合rt-PA组,每组又分在6 h、24 h、72 h、7 d四个时相,采用自身栓子法制备大鼠大脑中动脉栓塞模型(MCAO),rt-PA组及中药组分别于血栓注入后3 h一次性由尾静脉缓慢注入5.67mg/kg rt-PA,并分别给予高、常规、低剂量中药灌胃治疗(药量分别为:18ml/kg、9ml/kg、4.5ml/kg),每日2次。采用Western Blot法检测化痰通络法干预rt-PA溶栓后MCAO大鼠皮质中MMP-9蛋白表达变化的基础上,分别采用RT-PCR法与Western Blot法检测其对MMP-9上游激活物MMP-3、TIMP-1基因与蛋白表达的影响。结果与假手术组相比,模型组内各个时相大鼠大脑皮质梗死区MMP-9含量均显著增加,具有统计学意义(P<0.05);与模型组相比,rt-PA组MMP-9蛋白、MMP-3基因及蛋白表达量显著升高,而TIMP-1基因及蛋白表达呈下降趋势,差异具有显著性(P<0.05)。同一组内不同时相比较,MMP-9蛋白表达量6h最低,并随时间延长逐渐递增,以72h时MMP-9蛋白表达量最高,7d时MMP-9蛋白表达量呈下降趋势,差异均具有统计学意义(P<0.05);同一时相,与模型组比较,rt-PA组、中药低剂量组、中药中剂量组、中药高剂量组MMP-9的蛋白表达量皆有显著下降趋势(P<0.01);与rt-PA组比较,中药中剂量组和中药高剂量组MMP-9的蛋白表达量仍有显著下降趋势(P>0.05),虽无统计学意义,但在6h、1d时相以中药高剂量组降低MMP-9蛋白表达量更为明显,72h、7d时相则以中药中剂量组降低MMP-9的蛋白表达量趋势明显。同时与rt-PA组比较,中药中剂量组和中药高剂量组MMP-3表达均有下降(P<0.05),且TIMP-1表达亦有上升趋势。Objective To observe the effect of Huatan Tongluo on expression of MMP-3 and TIMP-1 on zymogen activation pathway of MMP-9 in rat cortex accepted thrombolysis therapy after the Acute Cerebral Infarction,to provide experimental evidence for the prevention of bleeding after thrombolysis in acute cerebral infarction with Huatan Tongluo. Methods 240 SD rats were randomly divided into five groups：sham group,model group,rt-PA group,Huatan Tongluo combined rt-PA low-dose group,Huatan Tongluo combined rt-PA middle-dose group and Huatan Tongluo combined rt-PA high-dose group（each group was divided in 6h,24 h,72 h,7 d four-phase）. MCAO rats were prepared by self-embolus method,rt-PA group and rt-PA Huatan Tongluo group use 5. 67 mg/kg rt-PA thrombolytic therapy after 3h thrombosis. The latter uses gavage treatment with low,medium,high dose of Huatan Tongluo prescription two times a day. Then respectively observe the expression of MMP-9 protein in MCAO rats cortex after thrombolysis in acute cerebral infarction,on this basic,observe the expression of MMP-3,TIMP-1 which are MMP-9 upstream activators by RT-PCR and Western Blot method. Results Compared with sham group,the expression of MMP-9 increased significantly（P〈0. 05）,Compared with model group,the expression of MMP-9 protein,MMP-3 gene and protein in rt-PA group were significantly higher and TIMP-1 was significantly lower（P〈0. 05）. Within the same group different time,the expression of MMP-9 lowest in 6 h,highest in 72 h,which was gradually increasing over time,the expression of MMP-9 was on declining curve in 7d,which differences are statistically significant（P〈0. 05）; In the same time,compared with model group,the expression of MMP-9 were all on declining curve in rt-PA group,Huatan Tongluo combined rt-PA low-dose group,Huatan Tongluo combined rt-PA middle-dose group and Huatan Tongluo combined rt-PA high-dose group（P〈0. 01）; compared with rt-PA group,the expression of MMP-9 were also on declining curve in Huatan Tongluo combined rt-PA mid

关 键 词：化痰通络法 急性脑梗死 RT-PA溶栓 MMP-9 活化途径 出血转化 

分 类 号：R285.5[医药卫生—中药学]