天然免疫细胞表面模式识别受体表达相互作用与-第三军医大学学报.DOCVIP

创伤感染时巨噬细胞表面模式识别受体表达、相互作用及其与炎症反应发生的关系 杨 策，陈永华，黄 宏，王海燕，谢国旗，蒋建新 (第三军医大学大坪医院野战外科研究所第四研究室,全军交通医学研究所,创伤、烧伤与复合伤国家重点实验室,重庆 400042) 摘 要： 从免疫细胞表面模式识别 选择小鼠内毒素血症、盲肠结扎穿孔、肺泡巨噬细胞免疫刺激模型，以免疫组织化学法和RT-PCR法检测主要模式识别受体的表达；利用细胞转染模型研究CD14、TLR4、MD2相互作用。 免疫细胞清道夫受体和CD14、TLR4、MD2，分别呈现下调和上调表达，且转染CD14、TLR4和MD2 3种表达质粒的HEK293细胞株对FITC-LPS结合率最强(P 0.05)，在内毒素刺激后NF-κB活性、TNF-α分泌最显著(P 0.05)。 免疫细胞表面防御性受体下调和效应性受体上调参与炎症反应失控过程，调控模式识别受体表达对减轻机体炎性损伤有重要意义。 关键词：模式识别受体; 内毒素; 创伤和损伤 中图法分类号：.1 文献标识码：AYANG Ce, CHEN Yong-hua, HUANG Hong, WANG Hai-yan, XIE guo-qi, JIANG Jian-xin (State Key Laboratory of Trauma, Burns and Combined Injury, Research Institute of Surgery, Research institute for Traffic Medicine of PLA, Daping Hospital, Third Military Medical University, Chongqing 400042, China) Abstract： Objective To investigate the possible mechanism of traumatic infection-related inflammation via the pattern recognization receptors (PRRs) in innate immunocytes and provide the new idea for the new measures in modulating the excessive inflammatory responses. Methods The models of endotoxemia mice, cecal ligation and puncture, and immmuno-stimulated alveolar macrophages were used for the assay of PRRs with immunohistochemistry and reverse transcription PCR method. HEK293 cells co-transfected plasmids expressing CD14, TLR4 and MD2 or two of them were stimulated with 100 ng/ml lipopolysaccharide (LPS) for the determination of NF-κB and tumor necrosis factor (TNF)-α. Results SR was up-regulated while the expression of CD14, TLR4 and MD2 were down-regulated in innate immunocyte. The HEK293 cells co-transfected three types of plasmids enhanced the sensitivity to LPS binding and stimulation compared with HEK293 cells co-transfected two of them (P0.05), followed with the significant translocation of NF-κB and TNF-α secretion (P0.05). Conclusion Our data strongly indicate that the significant correlation between uncontrolled inflammatory responses and the imbalance between defensive receptors and ef