苦柯胺B对脓毒症小鼠肺脏炎症反应的抑制作用  

Inhibitory effect of kukoamine B on lung inflammatory responses in mice with sepsis

作　　者：张锦丽[1] 秦魏婷 吕汪洄 沈唯长 王旭[1] 孙炳伟[1] 

机构地区：[1]江苏大学附属医院烧伤整形科,镇江212001

出　　处：《中华危重病急救医学》2014年第7期493-497,共5页Chinese Critical Care Medicine

基　　金：国家自然科学基金（30772256,81071546,81272148）;江苏省自然科学基金（BK2012703）

摘　　要：目的 探讨苦柯胺B（KB）对脓毒症小鼠肺部炎症反应的抑制作用及分子机制.方法 将28只雄性ICR小鼠按随机数字表法分为对照组（8只）、脂多糖（LPS）组（10只）、LPS ＋KB组（10只）.腹腔注射LPS 20 mg/kg制备脓毒症模型（LPS组）;对照组给予等体积生理盐水;LPS ＋KB组于LPS刺激4h后经尾静脉注射KB 20 μg/kg.于LPS刺激后8h检测动物血浆LPS浓度及肺组织髓过氧化物酶（MPO）活性;采用酶联免疫吸附试验（ELISA）检测血浆、肺泡灌洗液及肺组织匀浆中肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的含量;用蛋白质免疫印迹试验（Western Blot）检测肺组织核转录因子-κB（NF-κB）活性和诱导型一氧化氮合酶（iNOS）蛋白表达;用苏木素-伊红（HE）染色观察肺组织病理学改变;免疫组化法观察肺组织中细胞间黏附分子-1 （ICAM-1）蛋白表达.结果 与对照组比较,LPS组血浆LPS浓度（kEU/L：1 155.650±147.149比31.390±18.859）、MPO活性（U/g：1.177 ±0.093比0.775±0.166）、NF-κB活性（灰度值：1.557±0.105比0.824±0.032）、iNOS蛋白表达（灰度值：0.650±0.129比0.392±0.097）均显著升高（均P＜0.05）;KB干预后LPS浓度（624.461±149.012）、MPO活性（0.919±0.023）、NF-κB活性（1.127±0.074）、iNOS蛋白表达（0.425±0.066）均被明显抑制（均P＜0.05）.与对照组比较,LPS组血浆、肺泡灌洗液、肺组织匀浆中TNF-α（ng/L：47.325±13.864比6.534±0.544,13.382±2.231比3.748±0.692,31.127±7.399比14.948±4.673）、IL-1β（ng/L：74.329±11.890比29.921±6.487,9.422±2.674比1.105±0.364,528.509±32.073比109.945±13.561）浓度均显著增加（均P＜0.05）;而应用KB干预后TNF-α（20.331±7.789、7.145±1.202、15.966±2.946）、IL-1β（57.707±8.098、2.212±0.878、426.154±11.270）浓度均明显降低（血浆TNF-α：F=16.052、P=0.002,IL-1β：F=20.649、P=0.000;肺泡灌洗液TNF-α：F=31.134、P=0.001,IL-1β：F=22.792、Objective To investigate the inhibitory effect of kukoamine B （KB） on lung inflammatory responses in mice with sepsis and its possible molecular mechanism.Methods Twenty-eight male mice were randomly divided into control group （n=8）,lipopolysaccharide （LPS） group （n=10）,and LPS ＋ KB group （n=10）.Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg LPS,while equivalent normal saline was given in control group,and 20 μg/kg KB was injected through caudal vein 4 hours after LPS challenge in LPS ＋ KB group.After 8 hours of LPS challenge,the concentration of LPS in plasma and the activity of myeloperoxidase （MPO） in the lung tissue were determined.The contents of tumor necrosis factor-α （TNF-α） and interleukin-lβ （IL-1β） in plasma,alveolar lavage fluid and lung tissue homogenates were assessed by enzyme linked immunosorbent assay （ELISA）.The activation of nuclear factor-κB （NF-κB） and the expression of inducible nitric oxide synthase （iNOS） in lung tissue were determined by Western Blot.The pathological changes in lung tissues were observed with hematoxylin-eosin （HE） staining.The expression of intercellular adhesion molecule-1 （ICAM-1） in lung tissue was determined by immunohistochemistry.Results Compared with control group,the concentration of LPS in plasma （kEU/L：1 155.650 ± 147.149 vs.31.390 ± 18.859）,MPO activity （U/g：1.177 ±0.093 vs.0.775 ±0.166）,NF-κB activity （gray value：1.557 ±0.105 vs.0.824 ±0.032） and the expression of iNOS （gray value：0.650 ±0.129 vs.0.392 ±0.097） were significantly increased in LPS group （all P＜0.05）.After KB intervention,the concentration of LPS （624.461 ± 149.012）,MPO activity （0.919 ±0.023）,NF-κB activity （1.127 ±0.074） and the expression ofiNOS （0.425 ± 0.066） were significantly lowered （all P＜0.05）.Compared with control group,the contents of TNF-α （ng/L：47.325 ± 13.864 vs.6.534 ± 0.544,13.382 ± 2.231 vs.3.748 ± 0.692,31.127 ± 7.399

关 键 词：脓毒症 苦柯胺B 炎症反应 肺 

分 类 号：R459.7[医药卫生—急诊医学]