摘要
目的:研究蛭龙活血通瘀胶囊通过Ras同源基因/Rho相关螺旋卷曲蛋白激酶(Rho/ROCK)信号通路改善大鼠脑出血后脑水肿的分子机制。方法:将雄性SD大鼠随机分为模型对照组、蛭龙活血通瘀胶囊1.2 g/kg组、盐酸法舒地尔注射液12 mg/kg组,每组15只,自体血注入法复制脑出血模型,另设正常对照组,干预3 d后检测脑含水量及脑系数、血脑屏障通透性(EB含量)及超微结构,WB法检测脑组织闭锁小带蛋白(ZO-1)、咬合蛋白(Occludin)、RhoA、ROCK2、p-MLC蛋白表达,qRT-PCR法检测ZO-1、Occludin、RhoA、ROCK2 mRNA水平。结果:与正常对照组比较,模型对照组脑含水量及脑系数、脑组织EB含量、RhoA、ROCK2、p-MLC蛋白及RhoA、ROCK2 mRNA表达均明显升高(P<0.05或P<0.01),血脑屏障超微结构损坏明显,脑组织ZO-1、Occludin蛋白及mRNA表达均明显降低(P<0.05或P<0.01);与模型对照组比较,各药物组脑含水量及脑系数显著减少(P<0.01),脑组织EB含量显著降低(P<0.01),血脑屏障结构明显改善,ZO-1、Occludin蛋白表达显著上调(P<0.01),ZO-1、Occludin mRNA水平均明显上调(P<0.05),RhoA、ROCK2、p-MLC蛋白表达显著下调(P<0.01),RhoA、ROCK2 mRNA水平明显下调(P<0.05)。结论:蛭龙活血通瘀胶囊可能通过抑制Rho/ROCK通路调节血脑屏障功能,改善脑出血后脑水肿。
Objective:To research the molecular mechanism of Zhilonghuoxuetongyu Capsule on improving the brain edema of intracerebral hemorrhage in rats based on Rho/ROCK pathway. Methods:SD male rats were randomly divided into the control group, the model group, 1.2 g/kg Zhilonghuoxuetongyu Capsule group and 12 mg/kg Fasudil hydrochloride injection group, 15 rats in each group. The autologous blood injection was used to establish intracerebral hemorrhage model. 3 days after intervention, the brain water content and brain coefficient, blood-brain barrier permeability(EB content) and ultrastructure were detected, the protein expressions of ZO-1,occluding, RhoA,Rock2,P-MLC were detected by western blot method, and the mRNA expressions of ZO-1,occluding, RhoA and Rock2 were detected by qRT-PCR method. Results:Compared with the control group, the brain water content, brain coefficient, EB content, the protein and mRNA expressions of RhoA, Rock2, P-MLC and RhoA, Rock2 in the model group were significantly increased(P<0.01 or P<0.05). The ultrastructure of blood brain barrier was obviously damaged, and the protein and mRNA expressions of ZO-1, occludin in brain tissues were significantly decreased(P<0.01 or P<0.05). Compared with the model group, the brain water content and brain coefficient in each drug group were significantly decreased(P<0.01). The content of EB in the brain tissue was significantly decreased(P<0.01). The structure of blood-brain barrier was significantly improved. The protein expressions of ZO-1 and occludin were significantly increased(P<0.01),the mRNA levels of ZO-1 and occludin were significantly increased(P<0.05), the protein expressions of RhoA, Rock2 and P-MLC were significantly decreased(P<0.01), and the mRNA levels of RhoA and Rock2 were significantly decreased(P<0.05). Conclusion: Zhilonghuoxuetongyu Capsule may regulate the blood-brain barrier function by inhibiting Rho/ROCK pathway and improve the brain edema after intracerebral hemorrhage.
作者
王蔚
杜渊
王洪连
钟霞
孙长侦
刘增金
谭睿陟
杨思进
Wang Wei;Du Yuan;Wang Honglian;Zhong Xia;Sun Changzhen;Liu Zengjin;Tan Ruizhi;Yang Sijin(The Affiliated TCM Hospital of Southwest Medical University,Luzhou 646000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2021年第1期142-147,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金青年基金项目(编号:8170141602)
2018年泸州市酒城科技英才培育计划-中西医结合诊疗方案对出血性中风病的疗效研究
泸州市人民政府高端人才引进项目(刘新民团队)
四川省科技计划重点研发项目(编号:2019YFS0543)。
