弹性蛋白酶调节中性粒细胞炎性募集的功能研究  

Functional study on elastase regulation of inflammatory neutrophils recruitment

作　　者：刘洋[1] 孟玲 范思佳 任春光 张欢 LIU Yang;MENG Ling;FAN Sijia;REN Chunguang;ZHANG Huan(Laboratory of Developmental Biology,Department of Cell Biology and Genetics,School of Basic Medical Sciences,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学基础医学院细胞与遗传教研室发育生物学研究室,重庆400016

出　　处：《中国免疫学杂志》2024年第3期449-455,共7页Chinese Journal of Immunology

基　　金：中国博士后科学基金面上项目(2021MD703921);重庆市人社局重庆市留学人员回国创业创新支持计划(cx2020029);重庆市科委博士直通车项目(CSTB2022BSXM-JCX0036);重庆市博士后科学基金(cstc2021jcyj-bshX0080);重庆医科大学未来医学青年创新团队发展支持计划(W0052)。

摘　　要：目的:探索中性粒细胞弹性蛋白酶(NE)对中性粒细胞炎性募集的影响。方法:采用外源性弹性蛋白酶抑制剂西维来司钠盐预处理小鼠骨髓来源的中性粒细胞,通过腹膜炎过继性迁移、细胞体外趋化、流动室黏附、免疫荧光染色、细胞伸展等实验,检测NE的抑制对中性粒细胞体内炎性募集、体外趋化、牢固黏附、细胞极化与伸展能力的影响;利用流式细胞术及鲁米诺化学发光系统检测NE的抑制对中性粒细胞吞噬及活性氧(ROS)释放能力的影响。结果:西维来司钠盐预处理显著减弱了中性粒细胞体内炎性募集(P<0.001);减弱了中性粒细胞在体外趋化过程中对催化剂方向性的感知(P<0.05),减慢了趋化速率(P<0.05),降低了趋化距离(P<0.05);减少了中性粒细胞在炎性内皮细胞表面形成牢固黏附的能力(P<0.000 1),抑制了中性粒细胞对细菌的吞噬能力(P<0.01);但对中性粒细胞的伸展、极化及ROS的释放均无显著影响。结论:NE的抑制显著损害了中性粒细胞在体内外的炎性募集级联反应及吞噬作用,但对中性粒细胞的伸展、极化、ROS释放等功能无显著影响。Objective:To explore the effect of neutrophil elastase(NE)on neutrophil inflammatory recruitment.Methods:Mice bone marrow-derived neutrophils were pretreated with an exogenous elastase inhibitor-sivelestat sodium.The effects of elastase inhibition on the in vivo inflammatory recruitment,chemotaxis,adhesion,cell polarization and spreading of NE were examined by peritonitis adoptive transfer assay,dunn chamber,flow chamber,immunofluorescence staining and spreading assay,respectively.The effects of elastase inhibition on NE phagocytosis and reactive oxygen species(ROS)release capacity were detected by flow cytom-etry and the luminol chemiluminescence system.Results:Sivelestat sodium pretreatment significantly attenuated neutrophil in vivo in-flammatory recruitment(P<0.001);impaired neutrophil perception of chemotaxis in vitro(P<0.05),slowed chemotactic velocity(P<0.05),and decreased the chemotactic distance(P<0.05);reduced the adhesion of neutrophils to inflamed endothelial cells(P<0.0001)and inhibited the phagocytosis of bacteria by neutrophils(P<0.01);however,there was no significant effect on neutrophil spreading,polarization and ROS.Conclusion:NE inhibition significantly impaired the inflammatory recruitment cascade response and phagocyto-sis of neutrophils in vitro and in vivo but had no significant effect on the spreading,polarization and ROS release of neutrophils.

关 键 词：中性粒细胞 弹性蛋白酶 西维来司钠盐 炎性募集 

分 类 号：R392.12[医药卫生—免疫学]