Toll样受体在高迁移率族蛋白B1诱导严重烧伤后枯否细胞分泌促炎性细胞因子中的作用  

Role of toll-like receptor in high mobility group box protein 1-induced proinflammatory cytokines production from Kupffer cells in rats with severe burn

作　　者：陈旭林[1] 孙立[1] 郭峰[1] 王飞[1] 刘晟[1] 梁勋[1] 王仁素[1] 王永杰[1] 孙业祥[1] 

机构地区：[1]安徽医科大学第一附属医院烧伤科,安徽合肥230022

出　　处：《感染．炎症．修复》2015年第2期73-76,共4页Infection Inflammation Repair

摘　　要：目的:探讨Toll样受体(TLRs)在高迁移率族蛋白B1(HMGB1)诱导严重烧伤后枯否细胞(KCs)产生促炎性细胞因子TNF-α和IL-1β中的作用。方法:32只健康成年雄性SD大鼠随机分为两组。烫伤组大鼠采用背部置于98℃水中12 s的方法制成30%TBSAⅢ度烧伤模型并立即行液体复苏,假烫组以室温水替代98℃热水且不予液体复苏。伤后24 h,分离出肝脏KCs,分别以0、50、100、200 ng/ml HMGB1刺激48 h,ELISA法检测上清液中TNF-α和IL-1b含量。另将烫伤大鼠肝脏KCs随机分为对照组(正常培养48 h)、HMGB1组(100 ng/ml HMGB1刺激48 h)、HMGB1+TLR2/TLR4抗体组(以20μg/ml TLR2抗体或TLR4抗体培养2 h后加入100ng/ml HMGB1刺激48 h),ELISA法检测培养上清液中TNF-α和IL-1b含量,Northern blot法检测各组KCs中TNF-α和IL-1βm RNA的表达。结果:不同浓度HMGB1刺激严重烧伤及假烫大鼠KCs后,上清液中TNF-α和IL-1b含量呈剂量依赖性升高,且烧伤组水平明显高于假烫组(P<0.05或P<0.01);同时,KCs中TNF-α和IL-1b m RNA表达也显著升高。TLR2抗体和TLR4抗体均明显抑制HMGB1诱导的KCs产生TNF-α和IL-1b水平的升高,且TLR2抗体对HMGB1诱导的IL-1b升高的抑制作用强于TLR4抗体,而TLR4抗体对HMGB1诱导的TNF-α升高的抑制作用强于抗TLR2抗体。结论:TLR2和TLR4介导了HMGB1诱导严重烧伤大鼠KCs促炎性细胞因子TNF-α和IL-1b的表达。Objective： To investigate the role of toll-like receptor （TLR） in high mobility group box protein 1 （HMGB1）-induced production of proinflammatory cytokines, tumor necrosis factor-a （TNF-α） and interleukin-1β （IL-1β ）, from Kupffer cells （KCs） after severe burn injury in rats. Methods： Thirty-two healthy adult male Sprague- Dawley rats were randomly divided into two groups. In the scald group, model with full thickness burn injury involving 30% total body surface area （TBSA） was replicated in rats by immersing the dorsal skin in 98℃ water for 12 seconds followed by immediate resuscitation, In sham scald group, rats underwent sham burn in water of room temperature without fluid resuscitation. KCs were isolated from rats 24 hours after injury, and they were stimulated with HMGB1 （0, 50, 100, or 200 ng/ml） for 48 hours. The supernatant was collected for the determination of TNF-a and IL-lβ from KCs by enzyme-linked immunoadsorbent assay（ELISA）. The KCs from burned rats were divided into 4 groups randomly： （i） control group, KCs were left untreated and cultured for 48 hours; HMGB1 group, KCs were challenged with 100 ng/ml HMGB 1 for 48 hours; HMGBI＋anti-TLR2 antibody group, KCs were pre-incubated for 2 hours with anti-TLR2 monoclonal antibody and then challenged with 100 ng/ml HMGB1 for 48 hours; @ HMGBI＋ anti-TLR4 antibody group, KCs were pre-incubated for 2 hours with anti-TLR4 monoclonal antibody and challenged with 100 ng/ml HMGB1 for 48 hours. The mRNA expressions of TNF-a and IL-1β s were determined by Northern blot. Results： HMGB1 significantly increased TNF-a and IL-1β productions in a dose-dependent manner in supernatant of KCs from scald and sham scald rats. HMGB 1 stimulated more profuse production of TNF-a and IL- 1β by KCs from scald rats than those from animals of sham group （P〈0.05 or P〈0.01）. In addition, the expression of TNF-a and IL-β mRNAs were also significantly elevated in KCs after being stimulated by HMGB1. However, pre- incu

关 键 词：高迁移率族蛋白B1 烧伤 枯否细胞Toll 样受体 促炎性细胞因子 

分 类 号：R631.02[医药卫生—外科学]