百草枯中毒急性炎症反应中Tim-3的调控作用研究  

Roles of Tim-3 in paraquat poisoning induced acute inflammatory response

作　　者：于佳卉[1,2,3] 侯春梅[1] 蒋兴伟[1] 王位[1,2] 史青竹 陈国江 王仁喜[1] 肖鹤[1] 冯健男[1] 沈倍奋[1] 黎燕[1] 马远方[2] 韩根成[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]河南大学医学院免疫学研究所,河南开封475001 [3]河南省人口和计划生育干部学院,郑州450000

出　　处：《军事医学》2016年第1期45-50,共6页Military Medical Sciences

基　　金：卫计委公益行业科研专项资助项目(2015SQ000192)

摘　　要：目的在前期研究发现免疫调控蛋白Tim-3能负性调控炎症反应的基础上,探讨该分子能否通过调控免疫及炎症细胞稳态抑制百草枯(paraquat,PQ)中毒导致的急性炎症性损伤,以期为该类中毒性疾病的防治提供新的理论依据。方法制备PQ中毒小鼠模型,首先利用Tim-3转基因小鼠观察Tim-3高表达对PQ中毒后组织损伤及炎症介质、PQ血清浓度变化的影响;其次,制备Tim-3融合蛋白(s Tim-3),观察用该融合蛋白阻断Tim-3通路对PQ中毒后组织损伤及炎症介质变化的影响并探讨其机制。结果 PQ中毒后Tim3-转基因小鼠较野生型小鼠表现出肺及肾损伤减轻,同时Tim3-转基因小鼠肺内炎症介质TNF-α、IL-1β、IL-6的水平较野生型低。监测血清中的PQ浓度发现,Tim-3转基因小鼠血清中的PQ浓度显著低于对照组。而用Tim-3-trx融合蛋白阻断Tim-3信号后,PQ中毒小鼠的免疫反应及炎症损伤进一步加重。体外机制研究发现,Tim-3信号在减轻巨噬细胞炎症介质表达、维持炎症细胞稳态中发挥关键作用。结论在PQ中毒中,Tim-3对维持炎症反应稳态发挥关键作用,深入认识该信号通路的调控机制可望为减轻中毒后急性免疫组织损伤、降低患者死亡率提供新的策略。Objective To examine whether Tim-3 plays a protective role in paraquat poisoning induced excessive immune response and tissue damage based on the critical roles of Tim-3 controlling inflammatory response. Methods A paraquat poisoning model was established in wild type and in Tim-3 transgenic C57 BL /6 mice by intraperitoneal injection of paraquat（ 40 mg / kg）. In addition,C57 BL /6 mice with paraquat poisoning were injected with Tim-3 soluble protein（ s Tim-3）or control protein to see the effect of Tim-3 blocking on the progression of paraquat poisoning. Samples were collected at 6and 24 h after paraquat injection respectively and were examined for tissue damage,cytokine expression and paraquat metabolism. Results After paraquat poisoning,there was significantly attenuated tissue damage in the lungs and kidneys and decreased TNF-α,IL-6 and IL-1 beta expression in the PBMCs or in the serum from Tim-3 transgenic mice compared to wild type mice. The serum concentration of paraquat in Tim-3 transgenic mice was also significantly decreased. However,in s Tim-3 treated paraquat poisoning mice,there was significantly increased cytokine expression and tissue damage compared to control protein treated mice. The in vitro data showed that Tim-3 signaling negatively regulated macrophages mediated inflammatory response. Conclusion Tim-3 plays a critical role in maintaining the homeostasis after paraquat poisoning.Further investigation on the regulatory roles of Tim-3 in inflammation will shed new light on the pathogenesis of paraquat poisoning and provide new therapeutic strategies.

关 键 词：TIM-3 百草枯 中毒 炎症介质 肿瘤坏死因子Α 白细胞介素 

分 类 号：R595.4[医药卫生—内科学]