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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 method on gene expression of cerebral microvascular endothelial cells after thrombolysis of recombinant tissue plasminogen activator (rt-PA) in acute cerebral infarction rats, Experimental infarction thrombolysis after ischemia-reperfusion injury to provide experimental evidence. Methods: 240 healthy SD rats were randomly divided into 6 groups: Sham operation group, model group, rt-PA group, Huatan Tongluo Recipe low, medium and high dose rt-PA group Only the middle cerebral artery occlusion model (MCAO) was established by the auto embolectomy. The rt-PA thrombolysis was given to rt-PA group and Huatan Tongluo decoction combined with rt-PA group 3 h after thrombosis, The latter combination of low, medium and high doses (crude drug 3.6,7.2,14.4 g · kg-1) Huatan Tongluo intragastric administration, 2 times a day, respectively, 6,24,72 h after modeling, 7 At 4 d, the expressions of Occludin, claudin-1 and Zonula occludins protins-1 (ZO-1) genes in cortex and hippocampus of rats at different phases were observed by RTPCR. Results: At different time points, the expression of Occludin, claudin-1 and ZO-1 in different experimental groups were highest at 6 h, lowest at 24 h, and gradually increased at 72 h and 7 d Increasing trend more obvious. Compared with the 6 h group, the expression of Occludin, claudin-1 and ZO-1 in the rt-PA group was significantly higher than that in the low, medium and high dose groups 24 h and 72 h (P <0.05). At the same time, compared with the model group, the expression of Occludin and Claudin-1 in the medium and high dose groups were significantly increased (P <0.05). Although the expression of ZO-1 gene had similar trend, the difference was statistically significant only at 24, 72 and 7 d (P <0.05). Conclusion: Huatan Tongluo combined with rt-PA thrombolysis can regulate the expression of Occludin, claudin-1 and ZO-1 genes in microvascular endothelial cells to protect the integrity of blood-brain barrier and prevent and treat acute cerebral infarction The occurrence and development of ischemia reperfusion injury.