摘要
目的 从血脑屏障(blood-brain barrier,BBB)转运蛋白角度探讨冰片配伍黄芪甲苷(astragalosides IV,AST IV)和三七总皂苷(Panax notoginseng saponins,PNS)在脑缺血再灌注状态下促进药物成分入脑的作用。方法 采用大鼠局灶性脑缺血再灌注模型,ig给予冰片、AST IV、PNS及其配伍药物,2,3,5-氯化三苯四氮唑(2,3,5-triphenyl tetrazolium chloride,TTC)染色法测定脑组织损伤,Western blotting法检测脑组织外排蛋白P-糖蛋白(P-glycoprotein,P-gp)和多药耐药相关蛋白1(multidrug resistance protein-1,MRP-1)、MRP-2、MRP-4、MRP-5及摄取蛋白有机阳离子转运体3(organic cation transporter3,OCT3)、有机阴离子转运多肽-2(organicaniontransportingpolypeptides-2,OATP-2)蛋白的表达,real-timePCR法检测脑组织多药耐药(multidrug resistance,mdr)基因mdr1a、mdr1b和mrp-1、mrp-2、mrp-4、mrp-5 mRNA表达。结果 TTC染色结果显示,脑缺血再灌注后,大鼠脑组织出现明显梗死灶。各药物能显著减少脑梗死体积,ASTIV+PNS的效应强于AST IV与PNS单用,冰片+AST IV+PNS的效应强于各药物单用及AST IV+PNS。外排蛋白和基因检测显示,脑缺血再灌注后,大鼠脑组织P-gp、MRP-1、MRP-2、MRP-4、MRP-5蛋白表达均显著增多。与模型组比较,冰片能显著下调大鼠脑组织P-gp、MRP-2、MRP-4蛋白表达水平,PNS能显著下调MRP-4、MRP-5蛋白表达水平,AST IV、AST IV+PNS与冰片+AST IV+PNS能显著下调P-gp、MRP-2、MRP-4、MRP-5蛋白表达水平,且冰片+AST IV+PNS的效应显著强于各药物单用及AST IV+PNS,AST IV+PNS的效应显著强于AST IV或PNS单用。基因表达结果与蛋白表达结果类似。摄取蛋白检测结果显示,与对照组比较,脑缺血再灌注后大鼠脑组织OCT-3蛋白表达在模型组及各给药组变化均不明显,而模型组OATP-2蛋白表达水平显著降低。PNS、AST IV+PNS及冰片+AST IV+PNS能显著上调OATP-2蛋白表达水平,且AST IV+PNS的效应显著强于AST IV、PNS单用,冰片+ASTIV+PNS的效�
Objective To investigate the effects of borneol combined with astragaloside IV(AST IV)and Panax notoginseng saponins(PNS)on promoting the active components into the brain and anti-brain injury through the regulation of transporter proteins of blood-brain barrier(BBB)in the state of cerebral ischemia-reperfusion.Methods Focal cerebral ischemia-reperfusion model in rats was established,borneol,AST IV,PNS and the combination were administered by gavage,brain infarction rate was evaluated by 2,3,5-triphenyl tetrazolium chloride(TTC)staining,the expressions of efflux proteins such as p-glycoprotein(P-gp),multidrug resistance protein(MRP)-1,MRP-2,MRP-4,MRP-5 and uptake proteins such as organic cation transporter(OCT)-3,organicanion transporting polypep-tides(OATP)-2 in brain tissues were detected by Western blotting.The expressions of multidrug resistance(MDR)such as mdr1a,mdr1b and mrp-1,mrp-2,mrp-4,mrp-5 mRNA in brain tissues were determined by real-time PCR method.Results The results of TTC staining showed that brain infarct was found after cerebral ischemia-reperfusion.Each drug could significantly reduce brain infarction volume and decrease infarction rate,and the effect of AST IV+PNS was better than that of AST IV and PNS alone,the effect of borneol+AST IV+PNS was better than that of single drug and AST IV+PNS.The results of major efflux proteins and genes detection showed that the protein expressions of P-gp,MRP-1,MRP 2,MRP-4,and MRP-5 were significantly increased in rats after cerebral ischemia-reperfusion.Borneol could significantly down-regulate the expressions of P-gp,MRP-2,MRP-4 proteins,PNS could significantly down-regulate the levels of MRP-4,MRP-5 proteins;AST IV,AST IV+PNS and borneol+AST IV+PNS could significantly down-regulate P-gp,MRP-2,MRP-4,MRP-5 proteins,and the effects of borneol+AST IV+PNS were significantly better than those of single drug and AST IV+PNS;The effects of AST IV+PNS were significantly better than those of AST IV or PNS alone.The results of gene expressions were similar to those of
作者
唐三
丁煌
杨筱倩
陈仙蕾
黄小平
邓常清
TANG San;DING Huang;YANG Xiao-qian;CHEN Xian-lei;HUANG Xiao-ping;DENG Chang-qing(Molecular Pathology Laboratory,Key Laboratory of Hunan Province for Prevention and Treatment of Integrated Traditional Chinese and Western Medicine on Cardio-Cerebral Diseases,Key Laboratory of Hunan Universities for Cell Biology and Molecular Techniques,Hunan University of Chinese Medicine,Changsha410208,China)
出处
《中草药》
CAS
CSCD
北大核心
2019年第18期4389-4397,共9页
Chinese Traditional and Herbal Drugs
基金
湖南中医药大学“十三五”一级学科基础医学建设项目(No:06)
国家自然科学基金资助项目(81573875)
湖南省科技厅科技创新平台与人才计划-中医脑病临床研究中心(2017SK4005)
2017年度湖南省大学生研究性学习和创新性实验计划项目(1029)
“中医药防治心脑血管疾病基础研究”湖南省自然科学创新群体基金
关键词
冰片
黄芪甲苷
三七总皂苷
脑缺血再灌注
血脑屏障
转运蛋白
borneol
astragaloside IV
Panax notoginseng saponins
cerebral ischemia-reperfusion
blood-brain barrie
transporter proteins
