川芎嗪防治大鼠脑缺血/再灌注损伤后炎症反应与细胞凋亡的作用  

Effect of tetramethylpyrazine on inflammatory reaction and apoptosis after cerebral ischemia reperfusion injury in rats

作　　者：谢明 钟朕 欧阳训彦 郑延益 朱俊德 XIE Ming;ZHONG Zhen;OUYANG Xun-yan;ZHENG Yan-yi;ZHU Jun-de(Department of Human Anatomy,School of Basic Medical Sciences,Guizhou Medical University,Guizhou Guian 550025,China;Department of Infectious Diseases,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Orthopedics,the First Affiliated Hospital of Gannan Medical University,Jiangxi Ganzhou 341000,Chirm;Department of Nursing,Guizhou Forestry School,Guiyang 550201,Guiyang,China)

机构地区：[1]贵州医科大学基础医学院人体解剖学教研室,贵州贵安550025 [2]贵州医科大学附属医院感染科,贵阳550004 [3]赣南医学院第一附属医院骨科,江西赣州341000 [4]贵州省林业学校护理学教研室,贵阳550201

出　　处：《解剖学报》2021年第6期845-854,共10页Acta Anatomica Sinica

基　　金：国家自然科学基金(81660243);贵州省科技计划项目(黔科合基础-ZK[2021])一般415;贵州医科大学国家自然科学基金培育项目(20NSP006)。

摘　　要：目的探讨川芎嗪(TMP)防治大鼠炎症反应与细胞凋亡的作用机制。方法选取180只健康雄性SD大鼠,将其随机分为假手术组(sham)、脑缺血/再灌注损伤组(CIRI)、尼莫地平组(N)和TMP组,TMP再分为低剂量组(L)、中剂量组(M)和高剂量组(H) 3个亚组,CIRI组采用改良线栓法制备CIRI模型;TMP各组于术前30 min分别尾静脉注射5 mg/kg、10 mg/kg和30 mg/kg TMP进行干预,N组尾静脉注射尼莫地平(1 mg/kg),sham组和CIRI组给予等剂量的生理盐水。各组SD大鼠于构建CIRI模型苏醒后立即进行神经功能缺损评分,同时各组再灌注24 h进行2,3,5-氯化三苯基四氮唑(TTC)染色、苏木精-伊红(HE)染色及尼氏(Nissl)染色检测顶叶皮质缺血半暗带形态变化;ELISA检测顶叶皮质白细胞介素(IL)-1β及IL-8的表达,TUNEL检测顶叶皮质中的神经细胞凋亡,免疫荧光染色法检测β-catenin的阳性细胞在顶叶皮质表达,Western blotting检测顶叶皮质Bax和Bcl-2的表达。结果与sham组相比,CIRI组神经功能缺损评分显著增高(P<0.01),HE与Nissl染色见神经细胞肿胀变性,部分呈空泡样改变,细胞核固缩深染,神经元数量减少(P<0.01),尼氏体数量显著减少(P<0.01);炎症因子IL-1β和IL-8的浓度显著上升(P<0.01),凋亡细胞与β-连环蛋白(β-catenin)阳性细胞及其平均吸光度值均显著增多、增高(P<0.01);Bcl-2蛋白表达下降,而Bax蛋白表达显著增加(P<0.01)。与CIRI组比较,N组及TMP干预组大鼠的神经功能缺损评分降低(P<0.01),HE及Nissl染色发现大神经元水肿减轻,少许神经元被破坏,神经元数量增多(P<0.01),尼氏体数增多(P<0.01);炎症因子IL-1β和IL-8浓度显著下降(P<0.01),凋亡细胞与β-catenin阳性细胞及平均吸光度值均显著减少(P<0.01);Bcl-2蛋白表达增加,而Bax蛋白表达显著下降(P<0.01);与N组比较,随着TMP浓度的递增,神经功能、炎症反应与神经元病理变化呈现量效关系(P<0.05)。结论 TMP干预处理后可以减轻�Objective To explore the mechanism of tetramethylpyrazine( TMP) in preventing and treating inflammation and cell apoptosis in rats. Methods Totally 180 healthy male SD rats were selected and randomly divided into sham operation group( sham),cerebral ischemia reperfusion injury( CIRI) group,nimodipine group( nimodipine,N),and TMP subdivided into low-dose group( low). There were three subgroups: low-dose( L),medium dose( M),and high dose( H).In CIRI group a modified suture method was used to prepare the CIRI model;each TMP group was given tail injection 30 minutes before surgery. Intervention was given by intravenous injection of 5 mg/kg,10 mg/kg,and 30 mg/kg TMP. N group was given tail vein injection of nimodipine( 1 mg/kg),sham group and CIRI group were given the same dose of normal saline. SD rats in each group were scored for neurological deficits immediately after the CIRI model was constructed.At the same time,after 24 hours of reperfusion in each group,2,3,5-triphenyltetrazole chloride( TTC)staining,HE staining and Nissl staining were performed to detect the morphological changes of the parietal cortex ischemic penumbra;ELISA to detect the expression of IL-1β and IL-8 in the parietal cortex,TUNEL detects neuronal cell apoptosis in the parietal cortex,immunofluorescence detected the expression of β-catenin positive cells in the parietal cortex,and Western blotting detected the expression of Bax and Bcl-2 in the parietal cortex. Results Compared with the sham group,the neurological deficit score in the CIRI group was significantly higher( P < 0. 01). The HE and Nissl staining showed neuronal swelling and degeneration,some of which showed vacuole-like changes,pyknosis and deep staining of the nucleus,and a decrease in the number of neurons( P< 0. 01),the number of Nissl bodies was significantly reduced( P < 0. 01);the concentrations of inflammatory factors IL-1β and IL-8 increased significantly( P < 0. 01),apoptotic cells and β-cateninpositive cells and their average absorbance values both increased signifi

关 键 词：川芎嗪 脑缺血再灌注损伤 Β-连环蛋白 免疫印迹法 大鼠 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]