摘要
目的:探讨化痰通络方对重组组织纤溶酶原激活剂(rt-PA)溶栓后的急性脑梗死大鼠内质网应激(endoplasmic reticulum stress,ERS)后期神经细胞凋亡途径中Caspase-3蛋白以及Caspase-9蛋白表达的影响。方法:将健康160只SD大鼠随机分为假手术组、模型组、rt-PA组和实验组,每组各40只。采用自身栓子法以制备大鼠大脑中动脉栓塞模型(MCAO),rt-PA组与中药组分别给予SD大鼠尾静脉注入rt-PA(浓度:5.67 mg·kg-1)及联合化痰通络中药(浓度:7.2 g·kg-1)干预,,每日2次。每组分为6 h、1 d、3 d和7 d四个时相,并分别采用蛋白印迹法观察脑组织神经细胞中Caspase-9与Caspase-3蛋白的表达,采用TUNEL法观察各组大鼠脑神经元细胞的凋亡情况。结果:凋亡信号途径中Caspase-9蛋白以及Caspase-3蛋白的表达:与假手术组比较,模型组在4个时相中Caspase-9蛋白与Caspase-3蛋白表达量增加明显,差异具有统计学意义(P<0.05);而与模型组比较,rt-PA组和实验组在4个时相中Caspase-9蛋白与Caspase-3蛋白的表达量均降低差异具有统计学意义(P<0.05),与rt-PA组比较,实验组Caspase-9蛋白在1 d和7 d时表达量明显下低,Caspase-3蛋白在6 h、1 d、7 d时降低显著差异具有统计学意义(P<0.05)。结论:化痰通络法可以通过降低内质网应激后期神经细胞凋亡途径中Caspase-9与Caspase-3蛋白的表达,并且部分抑制神经细胞的凋亡,从而防治急性脑梗死溶栓后缺血再灌注的发生与发展。
Objective： To discuss the influence of Phlegm-Resolving and Collateral-Dredging Formula on the expression of Caspase-3 protein and Caspase-9 protein during nerve cell apoptosis in the late stage of ERS in rats with acute cerebral infarction after thrombolysis with rt-PA. Methods： Totally 160 healthy SD rats were randomly divided into sham-operation group,model group,rtPA group and experiment group,with 40 cases in each group. Middle cerebral artery occlusion model were made by self-embolus method,rt-PA group and Chinese medicine group. The SD rates were conducted caudal venous injection of rt-PA（ concentration：5. 67 mg·kg- 1） and combined with intervention of Phlegm-Resolving and Collateral-Dredging Chinese medicine（ concentration：7. 2 g · kg- 1）,for twice every day. The phases are respectively 6h,1d,3d and 7d for each group. And the expression of Caspase-3 protein and Caspase-9 protein was observed by the method of protein western blotting. The neuronal cells apoptosis in each group was observed by TUNEL method. Results： In the apoptosis signal pathway,the expression of Caspase-3 protein and Caspase-9 protein： compared with sham-operation group,the expression of the two proteins in model group in the four phases increased markedly,with statistical significance（ P 〈 0. 05）; compared with model group,the expression in of the two proteins the four phases in rt-PA group and experiment group declined,with statistical significance（ P 〈 0. 05）; compared with rt-PA group,the expression of Caspase-9 on the 1d and 7d got obvious decrease while the expression of Caspase-3 at 6h and on the 1d and 7d got statistically significant decrease（ P 〈 0. 05）. Conclusion： Phlegm-Resolving and Collateral-Dredging method can reduce the expression of Caspase-3 protein and Caspase-9 protein during nerve cell apoptosis in the late stage of ERS,thus preventing the occurrence and development of ischemia reperfusion after thrombolysis for acute cerebral infarction.
出处
《河南中医》
2016年第5期793-796,共4页
Henan Traditional Chinese Medicine
基金
国家自然科学基金项目(编号:81273942)
天津市应用基础及前沿技术研究计划(编号:12JZDJC25300)
天津市教委"十二五"创新团队培养计划资助(编号:TD12-5035)
