IL-1β等炎性因子在梭曼中毒大鼠脑内表达变化  

Changes in expression of IL-1β and other inflammatory cytokines in soman-intoxication rat brain

作　　者：宋尚华[1,2] 骆媛[2] 王永安[2] 

机构地区：[1]解放军总医院,北京100853 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《军事医学》2013年第9期661-665,670,共6页Military Medical Sciences

基　　金：国家科技重大专项资助项目(2011ZXJ09102-05C);全军医学科技"十二五"重大专项资助项目(AWS11C004)

摘　　要：目的观察梭曼染毒大鼠主要脑区炎性因子IL-1β、TNF-α及核转录调节因子NF-κB在不同时间的表达变化。方法建立大鼠梭曼致惊厥模型,采用HE染色法观察染毒后不同时段动物脑内梨状皮质及海马CA1区病理学改变;采用实时荧光定量PCR(RT-qPCR)及Western印迹测定上述两个脑区IL-1β、TNF-α及NF-κB mRNA和蛋白在不同时间的表达变化。结果病理学结果显示,梭曼中毒所致惊厥大鼠脑内梨状皮质及海马CA1区在中毒后30 min即出现明显病理学损伤,染毒后7 d仍持续存在。RT-qPCR测定结果发现,梨状皮质及海马CA1区内TNF-α、IL-1βmRNA在染毒后30 min表达明显增加,染毒后6 h达峰值,染毒后48 h仍大量表达;NF-κB mRNA表达变化相对较晚,在染毒后6 h出现显著上调,染毒后48 h表达仍异常增高。Western印迹结果表明,海马CA1区TNF-α及IL-1β的蛋白表达总体与mRNA表达类似,但梨状皮质TNF-α及IL-1β的蛋白表达变化总体出现较晚;而NF-κB的蛋白表达在梭曼中毒早期即出现显著升高。结论梭曼中毒大鼠主要脑区内炎性因子IL-1β、TNF-α、NF-κB mRNA及蛋白表达水平在中毒早期出现显著上调,提示针对炎性反应的干预措施可能成为梭曼中毒所致脑损伤的新治疗手段。Objective To observe changes of expression of inflammatory cytokines IL-1β,TNF-α and nuclear factor NF-κB in main areas of a rat＇s brain at different time points after soman poisoning.Methods After the establishment of the soman-induced seizure model in rats,the histopathological changes in the piriform cortex and CA1 of hippocampus at different time points following exposure to soman were observed by HE staining.Meanwhile,the levels of mRNA and protein of IL-1β,TNF-α and NF-κB were tested by real-time quantitative PCR and Western blot.Results（1） Neuropathological changes was shown at 30 min in the piriform cortex and CA1 of hippocampus following exposure to soman in convulsive rats.There was neuropathological damage at 7 d after poisoning.（2） The mRNA level of TNF-α and IL-1β increased significantly at 30 min,reached the top at 6 h and kept up-regulating until 7 d after soman poisoning in the piriform cortex and CA1 of hippocampus.The mRNA level of NF-κB remained unchanged until 6 h.The abnormal increase lasted until 48 h.（3） The protein expression of TNF-α and IL-1β changed like mRNA expressed in CA1 of hippocampus while the protein level in the piriform cortex changed slowly.The level of protein of NF-κB increased significantly in the early stage of seizure following exposure to soman.Conclusion The levels of mRNA and protein are up-regulated significantly in the early stage of seizure in the main areas of the brain of soman poisoned rats,suggesting that countermeasures focused on inflammatory reactions will be new therapeutic modality against soman induced brain damage.

关 键 词：梭曼 惊厥 脑损伤 炎性因子 

分 类 号：R827.17[医药卫生—临床医学] R994.7