EGCG对脑缺血再灌注损伤大鼠炎症反应的影响  

Effect of EGCG on inflammatory reaction in rats suffered cerebral ischemia/reperfusion injury

作　　者：贺芳[1] 叶蓓[1] 陈建珍[1] 李畅[2] HE Fang;YE Bei;CHEN Jianzhen;LI Chang(Department of Basic Medicine,Suzhou Vocational Health College,Suzhou Jiangsu 215009;Department of Thoracic Surgery,First Affiliated Hospital of Soochow University,Suzhou Jiangsu 215006,China)

机构地区：[1]苏州卫生职业技术学院基础医学教研室,江苏苏州215009 [2]苏州大学附属第一医院胸外科,江苏苏州215006

出　　处：《中南大学学报（医学版）》2021年第12期1325-1331,共7页Journal of Central South University ：Medical Science

基　　金：江苏省教育厅高校“青蓝工程”([2019]3);苏州市科技局科技发展计划项目(SYSD2018073);苏州卫生职业技术学院科技创新团队(SZWZYTD201806)。

摘　　要：目的:表没食子儿茶素没食子酸酯(epigallocatechingallate,EGCG)是茶叶中茶多酚生物活性的主要成分,具有多种生物学效应。脑缺血再灌注(ischemia/reperfusion,I/R)导致的神经损伤与炎症反应密切相关,促炎细胞因子白细胞介素-1β(interleukin-1β,IL-1β)与抗炎细胞因子白细胞介素-10(interleukin-10,IL-10)在炎症反应过程中起重要的调节作用。本研究旨在探讨EGCG对脑I/R损伤大鼠脑组织含水量、髓过氧化物酶(myeloperoxidase,MPO)活性以及IL-1β、IL-10的影响。方法:采用线栓法建立SD大鼠大脑中动脉栓塞模型,大鼠缺血1.5 h后,将尼龙线拔出,行再灌注24.0 h。将SD大鼠随机分成假手术(Sham)组、I/R组、EGCG1组(I/R+12.5 mg/kg EGCG)、EGCG2组(I/R+25.0 mg/kg EGCG)、EGCG3组(I/R+50.0 mg/kg EGCG)5组。Sham组大鼠除不予栓塞大脑中动脉外,其余与I/R组相同。采用干湿重法检测各组大鼠脑组织含水量,比色法检测MPO活性,RT-PCR测定IL-1β及IL-10 mRNA水平,ELISA测定IL-1β和IL-10含量。另取原代培养的SD大鼠大脑皮层神经元随机分成对照(Control)组、氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)组、OGD/R+EGCG组(OGD/R+50.0μmol/L EGCG)3组,采用蛋白质印迹法检测神经元IL-1β和IL-10蛋白质的表达水平。结果:与sham组比较,I/R组大鼠脑组织含水量、MPO活性、IL-1β含量、IL-10含量均显著增加(P<0.05或P<0.01),IL-1β及IL-10 mRNA表达水平均明显上调(均P<0.01)。与I/R组比较,EGCG3组大鼠脑组织含水量、MPO活性均显著降低(均P<0.01),IL-1β含量显著减少,IL-10含量显著增加(均P<0.01)。与I/R组比较,EGCG2组、EGCG3组大鼠缺血区脑组织IL-1βmRNA水平显著下调、IL-10 mRNA水平显著上调(P<0.05或P<0.01)。与Control组比较,OGD/R组神经元IL-1β及IL-10蛋白质表达水平均显著上调(均P<0.01)。与OGD/R组比较,OGD/R+EGCG组神经元IL-1β蛋白质表达明显下调,IL-10蛋白质表达明显上调(均P<0.01)。结论:EGCObjective:Epigallocatechin gallate(EGCG)is the main bioactive component of polyphenols in tea,which has a variety of biological effects.The neurologic injury caused by cerebral ischemia/reperfusion(I/R)is closely related to the inflammatory reaction.The pro-inflammatory factor interleukin-1β(IL-1β)and the anti-inflammatory factor interleukin-10(IL-10)play important roles in the regulation of inflammatory reaction.This study aims to explore the effect of EGCG on water content,myeloperoxidase(MPO)activity,IL-1βand IL-10 in brain tissues of rats suffered cerebral I/R injury.Methods:Middle cerebral artery occlusion model of SD rats was established by suture embolic method.After ischemia for 1.5 h,the nylon thread was pulled out and reperfusion was performed for 24.0 h.SD rats were randomly divided into 5 groups:a sham group,an I/R group,a EGCG1 group(I/R+12.5 mg/kg EGCG),a EGCG2(I/R+25.0 mg/kg EGCG),and a EGCG3 group(I/R+50.0 mg/kg EGCG).The sham group was the same as the I/R group except that the middle cerebral artery was not blocked.The water content in brain tissues of rats was measured by dry and wet weight method,and MPO activity was detected by colorimetry.The levels of IL-1βand IL-10 mRNA were determined by RT PCR,and the contents of IL-1βand IL-10 were determined by ELISA.In addition,primary cultured cortical neurons of SD rats were randomly divided into 3 groups:a control group,an oxygen-glucose deprivation/reperfusion(OGD/R)group,and an OGD/R+EGCG group(OGD/R+50.0μmol/L EGCG).The effects of EGCG on the levels of IL-1βand IL-10 protein in neurons were assessed byWestern blotting.Results:Compared with the sham group,water content,MPO activity,the contents of IL 1βand IL-10 were significantly increased(P<0.05 or P<0.01),the mRNA expression of IL 1βand IL-10 were obviously up-regulated(both P<0.01)in cerebral tissues of rats in the I/R group.Compared with the I/R group,water content and MPO activity in cerebral tissues of rats were significantly decreased(both P<0.01),the content of IL-1β(P<0.01)were

关 键 词：表没食子儿茶素没食子酸酯 缺血再灌注 炎症反应 白细胞介素-1Β 白细胞介素-10 

分 类 号：R743.3[医药卫生—神经病学与精神病学]