冰片对梓醇及葛根素透过局灶性脑缺血模型大鼠血脑屏障作用的研究  

Borneol promotes catalpol and puerarin crossing through blood-brain barrier in focal cerebral ischemic rats

作　　者：吴俊杰[1] 汪宏锦 杨帅[1] 刘杨[1] 徐晓玉[1] 

机构地区：[1]西南大学药学院&中医药学院中药研究所重庆市药效评价工程技术研究中心,重庆400715

出　　处：《中国中药杂志》2016年第21期3988-3995,共8页China Journal of Chinese Materia Medica

基　　金：国家级大学生创新创业训练计划项目(201510635017);国家自然科学基金项目(81473549);西南大学基本科研业务费专项(XDJK2014C089)

摘　　要：该研究通过观察冰片是否具有促进梓醇、葛根素透过局灶性脑缺血模型大鼠血脑屏障的作用,及其是否与激活β_2肾上腺素受体-eNOS-NO途径有关,从医学生物学作用角度阐释冰片"通窍引经"的传统功效及其机制。该研究采用大鼠大脑中动脉阻塞局灶性脑缺血模型,5 d后神经功能障碍评分达5分者为造模成功。将模型成功大鼠随机分为7组,用药前经统计学检验各组间无显著性差异后,灌服给予相应药物[梓醇45 mg·kg^(-1)、葛根素200 mg·kg^(-1)、冰片200 mg·kg^(-1)、布他沙明(Butoxamine,BTX)1.5 mg·kg^(-1)]:模型组(M组,溶剂)、梓葛组(ZG组)、梓葛冰组(ZGB组)、梓冰组(ZB组)、葛冰组(GB组)、β_2受体阻断剂+梓葛组(BTX+ZG组)、β_2受体阻断剂+梓葛冰组(BTX+ZGB组),另设假手术组(S组,溶剂),每组10只。各组给药10 min后取材,采用UPLC-MS测定脑脊液中梓醇和葛根素含量,Western blot法测定脑组织中eNOS含量、β_2受体表达,Griess法检测脑组织中NO含量。结果显示,与假手术组相比,造模大鼠神经功能得分均值均在5分以上,具有显著差异(P<0.05),显示造模成功。除假手术组外,经单因素方差分析,给药前各组之间的神经功能评分均无显著性差异。给药后与模型组相比,ZG组梓醇质量分数为26.673μg·L^(-1),葛根素含量在检测限以下,脑组织中eNOS表达和NO含量、β_2受体表达没有显著差异;与ZG组相比,ZGB组、ZB组、GB组脑脊液中梓醇质量分数增加至40~43μg·L^(-1)、葛根素质量分数增加至72~75μg·L^(-1),脑组织中eNOS表达和NO含量、β_2受体表达均有显著升高(P<0.05),显示冰片有促进作用;与ZG组相比,BTX+ZG组大鼠脑脊液中梓醇质量分数为21.401μg·L^(-1),葛根素检测不出,2组无显著差异,显示BTX无抑制作用;与ZGB组相比,BTX+ZGB组脑脊液中梓醇质量分数32.826μg·L^(-1)、葛根素质量分数47.820μg·L^(-1),脑组织中eNOS表达和NO含量均有显著下降(PPrevious studies showed that borneol could promote some drugs crossing through the blood-brain-barrier（ BBB） at certain conditions. However,the mechanism has not been clarified yet. This study aimed to investigate the effect of bornrol on promoting catalpol and puerarin through BBB and explore the relevant mechanism. The focal cerebral ischemic rats were divided into 7 groups randomly and then were administered corresponding drugs： model group（ M,solvent）,catalpol-puerarin group（ ZG,catalpol 45 mg·kg^（-1）＋ puerarin 200 mg·kg^（-1））,catalpol-puerarin-bornrol group（ ZGB,catalpol 45 mg·kg^（-1）＋ puerarin 200 mg·kg^（-1）＋ bornrol 200mg·kg^（-1））,catalpol-bornrol group（ ZB,catalpol 45 mg·kg^（-1）＋ bornrol 200 mg·kg^（-1））,puerarin-bornrol group（ GB,puerarin 200mg·kg^（-1）＋ bornrol 200 mg·kg^（-1））,butoxamine-ZG group（ BTX ＋ ZG,butoxamine 1. 5 mg·kg^（-1）＋ catalpol 45 mg·kg^（-1）＋ puerarin 200 mg·kg^（-1））,and butoxamine-ZGB group（ BTX ＋ ZGB,butoxamine 1. 5 mg·kg^（-1）＋ catalpol 45 mg·kg^（-1）＋ puerarin 200 mg·kg^（-1）＋ bornrol 200 mg·kg^（-1））. Another 10 sham-operated rats were set as control（ S）. Ten minutes after the administration,the cerebrospinal fluid was taken to test the content of catalpol and puerarin,and the brain tissue was taken to test the expression of β_2-adrenergic receptor,eNOS,and NO. Compared with the M group,the ZG group showed content of catalpol is 26. 673 μg·L^（-1）and the content of puerarin is below the detection limit; the expression levels of β_2-adrenergic receptor,eNOS and the contents of NO in brain tissue are no significant difference. Compared with the ZG group,the ZGB,ZB and GB groups showed significantly increased content of catalpol andpuerarin,as well as the expression of β_2-adrenergic receptor,eNOS and NO in the brain tissue（ P 0. 05）. The content of catalpol in BTX ＋ ZG group changed non-significantly. Compared with the ZGB group,the BTX �

关 键 词：冰片 梓醇 葛根素 血脑屏障通透性 Β2受体 eNOS NO 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]