p38丝裂原活化蛋白激酶信号转导通路在严重烧伤大鼠枯否细胞肿瘤坏死因子α和白细胞介素1β产生中的作用  

Role of p38MAPK signal transduction pathway in Kupffer cells production of TNF-α and IL-1β in severely burned rats

作　　者：陈旭林[1] 夏照帆[2] 韦多[2] 贲道锋[2] 王永杰[1] 

机构地区：[1]安徽医科大学第一附属医院烧伤科,合肥230022 [2]第二军医大学长海医院烧伤科

出　　处：《中华外科杂志》2005年第3期185-188,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金(30271340);国家 863课题(2001AA216041);军队"十五"指令性课题 (01L055 );上海市卫生系统百人计划课题(97BR046);安徽省教育厅科研基金(2004KJ207)

摘　　要：　目的　探讨p38丝裂原活化蛋白激酶(MAPK)信号转导通路在严重烧伤大鼠枯否细胞(KCs)促炎性细胞因子肿瘤坏死因子α(TNF -α)和白细胞介素 1β(IL -1β)产生中的作用。方法　健康成年的雄性SD大鼠 32只,随机分为:假烫组;假烫 +SB203580组;烧伤对照组;烧伤 +SB203580组,每组 8只。假烫或烧伤 24h后分离出肝脏KCs,培养 18h后加入 50ng/ml的LPS进行刺激, 18h后取上清液,用酶联免疫吸附法 (ELISA)测定TNF- α和IL- 1β的含量,并收集KCs,实时逆转录聚合酶链反应检测KCs内TNF -α和IL- 1βmRNA表达的改变,蛋白印迹 (Westernblot)法检测KCs中p38MAPK和JNK活性的变化。结果　烧伤大鼠分离出的KCs培养上清液中TNF -α和IL- 1β含量、KCs中TNF α和IL -1βmRNA的表达均较假烫组的明显增强,同时KCs中p38MAPK活性和JNK活性升高,SB203580能显著抑制大鼠KCs上清液中TNF -α和IL -1β含量、KCs中TNF- α和IL- 1βmRNA的表达和p38MAPK活性的升高,对JNK活性无影响。结论　p38MAPK信号转导通路介导了严重烧伤大鼠KCs促炎性细胞因子TNF- α和IL- 1β的产生。Objective To investigate the role of p38 mitogen-activated protein kinase (MAPK) signal transduction pathway in the Kupffer cells production of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in severely burns rats. Methods Male health adult Sprague-Dawley rats were randomized into four groups: sham burn rats given vehicle, sham burn rats given the p38 MAP kinase inhibitor SB203580, rats given a 30% total body surface area (TBSA) full-thickness burn and fluid resuscitation plus vehicle, and burn rats given injury and fluid resuscitation plus SB203580. Rats from each group were killed at 24 h after burn or sham burn and Kupffer cells (KCs) were isolated. After 18 h incubation, KCs next were stimulated with 50 ng/ml of LPS for 18 h. After stimulation, supernatants were removed for analysis of TNF-α and IL-1β levels by ELISA. The TNF-α and IL-1β mRNA expressions (by quantitative real-time RT-PCR) and the activities of p38 MAPK and JNK (by Western blot analysis ) in KCs were examined. Results Eighteen hours after 50 ng/ml LPS stimulation, KCs from burn rats released significantly higher levels of TNF-α and IL-1β than did shams. The mRNA levels of TNF-α and IL-1β in KCs increased significantly postburn. Western blot analysis suggested that expression of phosphorylated p38 MAPK and JNK were increased in KCs harvested from burn group after stimulation with LPS compared with those from sham group. In vivo administration of SB203580 markedly suppressed both the release of TNF-α and IL-1β and the mRNA expressions of TNF-α and IL-1β in KCs from both sham and burn rats. p38 MAPK activity in KCs was abolished by administration with SB203580, whereas JNK was not. Conclusions p38 MAPK signal transduction pathway mediates KCs production of proinflammatory cytokines TNF-α and IL-1β in severely burned rats.

关 键 词：IL-1β 肿瘤坏死因子α(TNF-α) 大鼠 严重烧伤 信号转导通路 白细胞介素1Β P38丝裂原活化蛋白激酶 RNA 雄性 MAPK 

分 类 号：R644[医药卫生—外科学] R692[医药卫生—临床医学]