化痰通络方对急性脑梗死大鼠rt-PA溶栓后不同脑区星形胶质细胞特异性蛋白表达的影响

Effects of Huatan Tongluo Decoction on Astrocytes Related Specific Protein Expression of Different Brain Tissues in Rats Which Use rt-PA Thrombolysis after Acute Cerebral Infarction

摘要:

目的：观察化痰通络方对急性脑梗死大鼠重组组织纤溶酶原激活剂（rt-PA）溶栓后不同脑组织中星形胶质细胞相关特异性蛋白水通道蛋白4（AQP-4）及血清中S100β蛋白表达的影响。方法：将240只健康SD大鼠随机分为6组：假手术组、模型组、rt-PA组、化痰通络方联合rt-PA组（中药低、中、高剂量组，剂量分别为3.6，7.2，14.4 g·kg^-1），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 5.67 mg·kg^-1 rt-PA溶栓治疗，后者联合给予低、中、高剂量化痰通络方中药灌胃治疗，6 h组于造模成功后给予中药1次；24 h组于造摸成功后及处死前2 h各给中药1次，共2次；72 h组于造模成功后开始给药，每日给中药2次，持续3 d，共6次；7 d组于造模成功后开始给药，每日给中药2次，持续7 d，共14次。分别于4个时相应用Western blot法观察不同时相大鼠皮质及海马AQP-4蛋白的表达，ELISA法检测血清中S100β蛋白的含量。结果：各实验组不同脑组织AQP-4相对丰度值及血清S100β含量自6 h起逐渐升高，24 h最高，随后逐渐降低，7 d时接近6 h水平；同一时相与假手术组比较，各实验组AQP-4及S100β蛋白均显著升高，说明造模成功。与模型组相比，中药中、高剂量组AQP-4蛋白于24 h或72 h或7 d时分别出现显著下降（P<0.05），研究结果并无明显集中现象，但整体趋势表明中药中、高剂量组能明显减少AQP-4的表达；rt-PA组、中药低、中、高剂量组S100β虽未出现明显差异，但均有不同程度降低，且以中药组下降趋势明显。结论：①溶栓治疗后，血脑屏障结构及功能损害于24 h时最为严重；②化痰通络方联合rt-PA溶栓可降低不同脑组织中AQP-4表达及血清中S100β含量进而防治急性脑梗死溶栓后出血转化的发生与发展。

Abstract:

Objective: To observe the effect of Huatan Tongluo decoction on aquaporin-4 (AQP-4) protein expression of different brain tissues and S100β of blood serum in rats which use rt-PA thrombolysis after the acute cerebral infarction. Method: SD rats 240 were randomly divided into six groups, sham operation group, model group, rt-PA group, rt-PA Huatan Tongluo of Chinese medicine low dose group (3.6 g·kg^-1), rt-PA Huatan Tongluo of Chinese medicine middle dose group (7.2 g·kg^-1), rt-PA Huatan Tongluo of Chinese medicine high dose group (14.4 g·kg^-1). Using the embolus method prepares the MCAO rat model. rt-PA group and rt-PA Huatan Tongluo group use rt-PA thrombolytic therapy after thrombosis. In 6 hours group the Chinese medicine were given one time, after making mode successfully.In 24 hours group, given two times, after making mode successfully and 2 hours were killed.In 72 hours, given six times, from making mode successfully, two times one day, lasting three days.In 7 day group;given fourteen times, from making mode successfully, two times one day, lasting seven days. Then respectively observe the AQP-4 protein expression in rat cortex and hippocampus by Western blot method, and S100β of blood serum by ELISA method at 6, 24, 72 h, 7 days. Result: Every experimental group in different phases, the AQP-4 protein expression in different brain tissue and S100β of blood serum were highest at 6, 24 h maximum, 72 h, 7 d increased gradually to the level of 6 h. Compared with the model group at the same time the AQP-4 protein expression of middle dose group and high dose group of Chinese traditional medicine were significantly decreased at 24 h or 72 h or 7 d (P<0.05). There was no obvious concentration, but the overall trend showed that Chinese medicine middle dose group and high dose group can obviously decrease the expression of AQP-4.The S100β protein expression didn't show any significant differences in any group, but all have reduced in different degree, and the downward trend is more obvious in Chinese traditional medicine groups. Conclusion: The blood brain barrier structure and function was damaged most seriously at 24 h phase after thrombolysis treatment.Huatan Tongluo combined with rt-PA therapy can reduce AQP-4 expression of different brain tissues and S100β content of serum. So that it can protect the hemorrhagic transformation after thrombolytic treatment of acute cerebral infarction.