三化汤对脑缺血再灌注损伤大鼠血脑屏障的保护作用  

Protective Effect of Sanhua Decoction on Blood-brain Barrier in Rats with Cerebral Ischemia-reperfusion Injury

作　　者：李雯洁 巩子汉 黄颖[1] 孙明杰[1] LI Wenjie;GONG Zihan;HUANG Ying;SUN Mingjie(Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院医学实验中心,北京100700 [2]中国中医科学院中医基础理论研究所,北京100700

出　　处：《中国中医基础医学杂志》2022年第7期1072-1076,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：北京市自然科学基金面上项目(7192142)-基于脑微血管“分子开关”KLF2探讨祛风药羌活在三化汤治疗脑卒中的增效作用机理;中央级公益性科研院所基本科研业务费专项资金(ZZ2018005)-基于脑微血管调节作用的祛风药羌活在三化汤中治疗缺血性中风机理研究。

摘　　要：目的:研究经典名方三化汤对脑缺血再灌注损伤(cerebral ischaemia-reperfusion injury,CIRI)大鼠血脑屏障(blood-brain barrier,BBB)的保护作用及其组方配伍机制。方法:将SD大鼠分为正常组、假手术组、造模组,造模组采用Longa线栓法建立脑缺血再灌注损伤大鼠模型,造模成功后随机分为模型组、三化汤组、三化汤去羌活组(简称去羌活组)、羌活组、尼莫地平组各14只。灌胃相应药物5 d后股静脉注射2%伊文思蓝(evans blue,EB)溶液检测BBB通透性,Western blot和荧光定量PCR检测脑组织Krüppel样因子2(krüppel-like factor 2,KLF2)、血栓调节蛋白(Thrombomodulin)和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)的蛋白和mRNA表达。结果:与正常组比较,模型组大鼠脑组织EB含量显著升高(P<0.01),KLF2蛋白与mRNA表达均显著下降(P<0.05,P<0.01),Thrombomodulin、eNOS蛋白与mRNA表达均显著上升(P<0.01);与模型组比较,三化汤组、羌活组和尼莫地平组大鼠脑组织EB含量、Thrombomodulin、eNOS的蛋白与mRNA表达均显著下降(P<0.05,P<0.01);与模型组比较,三化汤组、羌活组KLF2蛋白与mRNA表达均显著上升(P<0.05,P<0.01)。结论:三化汤对脑缺血再灌注大鼠BBB损伤具有保护作用,这种保护作用是通过调控KLF2、Thrombomodulin、eNOS的表达以保护内皮完整性、调节血管内皮功能实现的,祛风药羌活可能在三化汤保护CIRI大鼠BBB过程中发挥了重要作用。Objective:To explore the mechanism of the protective effects of Sanhua decotion on blood-brain barrier(BBB)in cerebral ischaemia-reperfusion injury(CIRI)rats and the compatible characteristics.Methods:Firstly,SD rats were randomly divided into normal group,sham-operation group,operation group.Using Longa suture method to establish the CIRI model.The successful CIRI model rats were divided into model group,Sanhua decoction group,Sanhua decotion without Qianghuo group(named as without Qianghuo group),Qianghuo group,and Nimodipine group,with 14 rats in each group.Distilled water or medicine was administrated for 5 days before 2%Evans blue dye was administrated via the femoral vein to evaluate the BBB permeability.The brain tissues were used to detect KLF2,Thrombomodulin,eNOS expression by RT-qPCR and Western blot methods.Results:Compared with normal group,the EB content increasd significantly(P<0.01),the expression of KLF2 protein and mRNA decreased significantly(P<0.05,P<0.01),and the expressions of eNOS,Thrombomodulin protein and mRNA increasd significantly(P<0.01)in the brain tissue of the model group.Compared with model group,the contents of EB,the expression of thrombomodulin,eNOS protein and mRNA decreased significantly in the brain tissue of rats in the Sanhua decoction group,Qianghuo group and Nimodipine group(P<0.05,P<0.01).Compared with model group,the expression of KLF2 protein and mRNA increased significantly in the brain tissue of rats in the Sanhua decoction group and Qianghuo group(P<0.05,P<0.01).Conclusion:Sanhua decoction exerted a strong protective effect on BBB in CIRI rats by regulating the expression of KLF2,eNOS,and thrombomodulin to protect vascular integrity and endothelial function.The wind-dispelling drug Qianghuo may play an important role in the protective effects of Sanhua decoction on BBB in CIRI rats.

关 键 词：三化汤 羌活 祛风药 脑缺血再灌注 血脑屏障 

分 类 号：R285.5[医药卫生—中药学]