化痰通络方对急性脑梗死大鼠溶栓后脑微血管内皮细胞构成蛋白基因表达的影响  

Effects of Huatan Tongluo on Gene Expression of Brain Microvascular Endothelial Cells in Rats with Acute Cerebral Infarction after Thrombolysis

作　　者：张玉莲[1] 周震[1] 刘爽[1] 宋宛珊[2] 马妍[2] 田涛涛[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：《中国实验方剂学杂志》2014年第2期157-162,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072796);天津市应用基础及前沿技术研究计划(10JCZDJC20500)

摘　　要：目的:观察化痰通络法对急性脑梗死大鼠重组组织型纤溶酶原激活剂(rt-PA)溶栓后脑微血管内皮细胞构成蛋白基因表达的影响,为化痰通络法防治急性脑梗死溶栓后缺血再灌注损伤提供实验依据。方法:240只健康SD大鼠随机分为6组:假手术组、模型组、rt-PA组、化痰通络方低、中、高剂量联合rt-PA组(联合组),每组40只,采用自身栓子法制备大鼠大脑中动脉栓塞模型(MCAO),rt-PA组与化痰通络方联合rt-PA组于血栓注入后3 h一次性予以rt-PA溶栓治疗,后者联合给予低、中、高剂量(生药3.6,7.2,14.4 g·kg-1)化痰通络方灌胃治疗,每日2次,分别于造模后6,24,72 h,7 d 4个时点,应用RTPCR法观察不同时相大鼠皮质及海马紧密连接蛋白Occludin,claudin-1,Zonula occludins protins-1(ZO-1)基因的表达。结果:在不同时点,各实验组在不同脑组织中Occludin,claudin-1,ZO-1基因表达量均在6 h最高,24 h最低,72 h,7 d数值逐渐升高,且联合组升高趋势更明显。与组内6 h时比较,rt-PA组、联合低、中、高剂量组Occludin,claudin-1,ZO-1基因表达量多于24,72h(P<0.05)。而同一时点与模型组相比,联合中、高剂量组各时点Occludin,claudin-1基因表达量均明显升高(P<0.05)。而ZO-1基因表达量虽具有相似变化趋势,但仅在24,72 h,7 d差异具有统计学意义(P<0.05)。结论:化痰通络方联合rt-PA溶栓,可通过调控微血管内皮细胞构成蛋白Occludin,claudin-1,ZO-1基因的表达,进而保护血脑屏障完整性,防治急性脑梗死溶栓后缺血再灌注损伤的发生与发展。Objective： To observe the effect of Huatan Tongluo therapy on gene expression of brain microvascular endothelial cells in rats with acute cerebral infarction after recombinant tissue-type plasminogen（rt-PA） thrombolysis, and provide experimental basis for preventive and therapeutict effects of Huatan Tongluo on ischemia-reperfusion injury induced by acute cerebral infarction after thrombolysis. Method： Two hundred and fouty healthy SD rats were randomly divided into six groups, sham operation group, model group, rt-PA group, rt-PA combined with Huatan Tongluo low dose group, rt-PA combined with Huatan Tongluo middle dose group, rt-PA combined with Huatan Tongluo high dose group. The middle cerebral artery occlusion（MCAO） rat model was prepared by autologous blood clots. rt-PA group and rt-PA combined with Huatan Tongluo groups used rt-PA thrombolytic therapy after thrombosis. The later were also given low, medium, high dose（3.6, 7.2, 14.4 g·kg-1） Huatan Tongluo prescription by gavage two times a day. Then observed the expression of occluding, claudin-1, ZO-1 in rat cortex and hippocampus by RT-PCR at 6, 24, 72 h and 7 days respectively. Result： In different phases of every experimental group, the expression of occluding, claudin-1, ZO-1 in different brain tissue were highest at 6 h, minimum at 24 h, increased gradually at 72 h and 7 days, and the combined with Huatan Tonglow groups increased more obviously. The expression of occluding, claudin-1, ZO-1 in rt-PA, low, middle, high dose groups at 6 h were significantly higher than that at 24, 72 h（P〈0.05）. Compared the model group at the same time, the expression of occluding, claudin-1 in middle dose and high dose groups of Chinese traditional medicine were significantly increasedin each phase （P〈0.05）. While the ZO-1 gene expression had the similar trends, but significant difference only existed at the 24, 72 h, 7 d （P〈0.05）. Conclusion： Huatan Tongluo combined with rt-PA therapy can regulate the expression of occluding, clau

关 键 词：化痰通络法 急性脑梗死 OCCLUDIN CLAUDIN-1 ZO-1 

分 类 号：R285.5[医药卫生—中药学]