黄芪和三七的主要有效成分配伍对脑缺血/再灌注小鼠NF-κB信号通路及炎性因子表达的影响  

Effects of main active component combinations between Astragalus and Panax notoginseng on NF-κB signaling pathway and expressions of inflammatory factors after cerebral ischemia-reperfusion in mice

作　　者：黄小平[1] 卢金冬[1] 丁煌[1] 邓冰湘[1] 唐映红[1] 邓常清[1] 

机构地区：[1]湖南中医药大学分子病理实验室,中西医结合心脑疾病防治湖南省重点实验室,细胞生物学与分子技术湖南省高校重点实验室,湖南长沙410208

出　　处：《中国药理学通报》2015年第1期141-146,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81102557);教育部2010年度高等学校博士学科点专项科研基金资助项目(No20104323110001);湖南省高校创新平台开放基金资助项目(No 11K050,14K068);湖南省科技厅一般项目(No2014SK3001);湖南省中医药管理局重点项目(No201301);湖南省教育厅一般项目(No 11C0963);“中西医结合防治心脑血管疾病的相关基础研究”湖南省高校科技创新团队;“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

摘　　要：目的从炎症反应探讨黄芪和三七主要有效成分配伍抗脑缺血/再灌注损伤的机制。方法将C57BL/6小鼠随机分为假手术组、模型组、黄芪甲苷(ASTⅣ)组、人参皂苷Rg1(Rg1)组、人参皂苷Rb1(Rb1)组、三七皂苷R1(R1)组、4种有效成分配伍组、ASTⅣ+Rg1组、ASTⅣ+Rb1组、ASTⅣ+R1组及阳性对照药依达拉奉组。预先给药3 d,末次给药1 h后,结扎双侧颈总动脉,造成脑缺血20 min,再灌注24h。RT-PCR法测定TNF-α、IL-1β、ICAM-1 mRNA表达,Western blot法测定脑组织p-IκBα蛋白及胞质、胞核NF-κB蛋白表达,计算NF-κB核转位率。结果 1脑缺血/再灌注后,脑组织TNF-α、IL-1β、ICAM-1 mRNA表达增加。ASTⅣ可降低TNF-αmRNA表达,Rg1可降低ICAM-1 mRNA的表达,R1能同时下调TNF-α、ICAM-1 mRNA的表达。ASTⅣ与Rg1、Rb1、R1配伍对TNF-α、IL-1β、ICAM-1 mRNA的表达均有不同程度的抑制作用,且以4种有效成分配伍组和ASTⅣ+R1组对炎性细胞因子表达的抑制效应更加明显。2脑缺血/再灌注后,脑组织p-IκBα蛋白表达明显增加,胞质NF-κB蛋白表达明显减少,而胞核NF-κB蛋白表达增加,NF-κB核转位率升高。ASTⅣ、Rg1、R1及各有效成分配伍均能抑制IκB磷酸化,减少NF-κB核转位的发生,且配伍组的效应大于各有效成分单用,4种有效成分配伍组的效应大于ASTⅣ+Rb1和ASTⅣ+Rg1组。结论黄芪和三七的主要有效成分配伍能通过抑制缺血/再灌注后脑组织NF-κB信号通路的激活,减少炎性细胞因子的生成,从而减轻脑缺血后脑组织的继发性炎症反应,发挥对脑组织的保护作用。Aim To probe the possible mechanisms of the main active component combinations between As-tragalus and Panax notoginseng antagonizing on ische-mia-reperfusion injury from the perspective of the in-flammatory response. Methods C57BL/6 mice were randomly divided into the following group：sham, mod-el, Astragaloside Ⅳ （ AST Ⅳ） , Ginsenoside Rg1 （ Rg1 ） , Ginsenoside Rb1 （ Rb1 ） , Notoginsenoside R1 （R1）, four active component combination, AST Ⅳ＋Rg1, AST Ⅳ ＋Rb1, AST Ⅳ ＋R1 and Edaravone, pretreated for 3 d. After 1 h of the last administration, the model of cerebral ischemic-reperfusion injury was established by bilateral common carotid artery （ CCA） ligation for 20 min followed by reperfusion for 24 h. The expression of TNF-α, IL-1β, ICAM-1 mRNA was detected by RT-PCR, the expression of p-IκBαas well as NF-κB proteins of cytoplasm and nucleus in brain tissues was tested by Western blot,and NF-κB nuclear translocation rate was caculated. Results ①After cerebral ischemia-reperfusion, the expressions of TNF-α, IL-1β, ICAM-1 mRNA in brain tissues were up-regulated. AST Ⅳ could down-regulate TNF-α mRNA expression, Rg1 could down-regulate ICAM-1 mRNA expression, R1 could decrease both TNF-αand ICAM-1 mRNA expressions, and AST Ⅳ combined with Rg1 , Rb1 , R1 had different degrees of inhibitions on TNF-a, IL-1β, ICAM-1 mRNA. Moreover, the inhibi-tory effects of four active component combination and AST Ⅳ＋R1 on the inflammatory cytokines were more obvious. ②After cerebral ischemia-reperfusion, the expression of p-IκBα protein in brain tissues was sig-nificantly increased, NF-κB protein was significantly down-regulated in cytoplasm while up-regulated in nu-cleus, and nuclear translocation rate raised. AST Ⅳ, Rg1 , R1 and the active component combinations could inhibit the phosphorylation of IκB, reduce the occur-rence of NF-κB nuclear translocation, furthermore, the effects of the combination were better than those of the active components alone, and the effects

关 键 词：黄芪 黄芪甲苷 三七 人参皂苷 RG1 人参皂苷Rb1 三七皂苷R1 有效成分配伍 脑缺血/再灌注 信号通路 炎性细胞因子 

分 类 号：R-332[医药卫生] R282.71