异氟醚对局灶性脑缺血/再灌注大鼠血管生成及Smad信号通路的影响  

Effects of isoflurane on angiogenesis and Smad signaling pathway in rats with focal cerebral ischemia/reperfusion

作　　者：彭莉 王胜[2] 殷姜文 葛明月 张贵星 徐锋 张庆桐 PENG Li;WANG Sheng;YIN Jiang-wen;GE Ming-yue;ZHANG Gui-xing;XU Feng;ZHANG Qing-tong(Dept of Anesthesiology,the First Affiliated Hospital of Henan University,Kaifeng Henan 475000,China;Dept of Anesthesiology,the First Affiliated Hospital,University of Science and Technology of China,Hefei 230001,China;Dept of Anesthesiology,the First Affiliated Hospital of Shihezi University,Shihezi Xingjiang 832000,China)

机构地区：[1]河南大学第一附属医院麻醉科,河南开封475000 [2]中国科学技术大学附属第一医院麻醉科,安徽合肥230001 [3]石河子大学医学院第一附属医院麻醉科,新疆石河子832000

出　　处：《中国药理学通报》2019年第12期1731-1738,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81860249,81360203)

摘　　要：目的探讨异氟醚对脑缺血/再灌注大鼠血管生成的影响及其可能机制。方法健康成年♂SD大鼠40只,随机分为假手术组(Sham组)、缺血/再灌注组(I/R组)、异氟醚后处理组(ISO组)和异氟醚后处理+Smad3特异性抑制剂SIS3 HCl组(ISO+SIS3组)。线栓法建立大鼠大脑中动脉栓塞模型(MCAO),24 h后Zea-Longa法评估大鼠神经功能损伤,HE染色评估脑组织病理学损伤情况,尼氏染色评估缺血脑组织中健存神经元情况,TUNEL法评估脑组织凋亡情况,免疫荧光检测脑组织中VEGF蛋白和CD34蛋白的表达水平,Western blot对脑组织中p-Smad3、Smad3、VEGF和CD34进行定量分析。结果异氟醚能明显降低大鼠的神经行为学评分,减轻脑组织病理学损伤,增加缺血脑组织中健存神经元数量,减少损伤脑组织中的凋亡细胞,增强p-Smad3、VEGF和CD34蛋白的表达;Smad3抑制剂逆转了异氟醚的脑保护作用,加重了脑缺血/再灌注损伤,抑制p-Smad3、VEGF和CD34的蛋白表达。结论异氟醚能改善大鼠脑缺血/再灌注损伤,其保护机制与激活Smad信号通路,促进VEGF和CD34蛋白表达,进而促进血管生成有关。Aim To investigate the effects of isoflurane on angiogenesis in rats with cerebral ischemia/reperfusion and the possible mechanism.Methods Forty healthy adult male Sprague-Dawley rats were randomly divided into sham operation group(Sham group),ischemia-reperfusion group(I/R group),isoflurane post-treatment group(ISO group)and isoflurane post-treatment+Smad3 specific inhibitor SIS3 HCl group(ISO+SIS3 group).Rat middle cerebral artery occlusion model(MCAO)was established by suture method.After 24 h,Zea-Longa method was used to evaluate the neurological deficit of rats.HE staining was used to evaluate the pathological damage of brain tissues.Nissl staining was used to evaluate the surviving neurons in ischemic brain tissues.TUNEL staining was employed to assess the apoptosis of brain tissues.Immunofluorescence was applied to evaluate the expression levels of VEGF and CD34.Western blot analysis was used to detect the expression levels of p-Smad3,Smad3,VEGF and CD34.Results Isoflurane significantly reduced the neurobehavioral score of rats,reduced the pathological damage of brain tissues,increased the number of normal neurons in the ischemic brain tissues,reduced the apoptotic cells in injured brain tissues,and enhanced the expression levels of p-Smad3,VEGF and CD34.Smad3 inhibitor reversed the brain protective effect of isoflurane,aggravated cerebral ischemia-reperfusion injury,and inhibited the protein expression levels of p-Smd3,VEGF and CD34.Conclusions Isoflurane can improve cerebral ischemia/reperfusion injury in rats,and its protective mechanism is related to activation of Smad signaling pathway,promotion of VEGF and CD34 protein expression,and promotion of angiogenesis.

关 键 词：异氟醚 后处理 局灶性脑缺血/再灌注 血管生成 SMAD3 信号通路 

分 类 号：R-332[医药卫生] R322.81