香烟烟雾诱导IL-17A缺失/减弱小鼠的肺部炎性反应及其机制研究

Cigarette Smoke Induce IL-17A Deficiency/attenuation in Lung Inflammatory Response in Mice and Its Mechanism

目的探讨白细胞介素-17A(IL-17A)在香烟烟雾(CS)肺损伤中的作用及其机制。方法分别将IL-17A敲除(IL-17A-/-)小鼠、WT(野生型,C57BL/6J)小鼠、施用IL-17A中和抗体滴鼻的BALB/c小鼠、BALB/c小鼠以及非肥胖糖尿病/重症联合免疫缺陷(NOD SCID)小鼠,各种小鼠均设烟雾组(CS组)和Sham组;CS组置于CS环境中诱导2周。梯度离心小鼠肺泡灌洗液分离小鼠肺中的NK/NKT细胞、巨噬细胞和中性粒细胞并计数,同时用qPCR的方法检测肺组织IL-17A及其他相关基因的转录水平。结果CS诱导下IL-17A敲除小鼠肺泡灌洗液中的总细胞数、巨噬细胞数和中性粒细胞数均低于WT小鼠,肺组织CCL2、CCL3和MMP-12的转录水平均低于WT小鼠(P均<0.01);CS诱导下IL-17A单克隆抗体滴鼻处理的BALB/c小鼠肺泡灌洗液中的总细胞数、巨噬细胞数和中性粒细胞数均低于Sham组小鼠,CCL2、CCL3和MMP-12的转录水平均低于Sham组小鼠(P均<0.01);CS诱导NOD SCID小鼠的巨噬细胞、中性粒细胞和NK/NKT细胞代偿性表达IL-17A。结论CS诱导小鼠模型中IL-17A缺失/减弱可减轻肺部炎性反应,并且CS诱导下IL-17A可以由小鼠肺部T细胞以外的巨噬细胞、中性粒细胞等分泌。

Objective To observe the effect of interleukin-17A knockdown or reduction on the mouse cigarette smoke(CS)induction model and its mechanism.Methods IL-17A-/-mice,WT(wild type,C57BL/6J)mice,BALB/c mice with IL-17A neutralizing antibody nasal drip,BALB/c mice and non-obese diabetes/severe combined immunodeficiency(NOD SCID)mice were divided into smoke group(CS group)and Sham group respectively.CS group was induced in CS environment for 2 weeks.Gradient centrifugation was used to isolate NK/NKT cells,macrophages and neutrophils from the lungs of mice.The transcription levels of IL-17A and other related genes were detected by qPCR.Results The total number of cells,macrophage and neutrophils in alveolar lavage fluid of IL-17A knockout mice induced by CS were lower than those of wild type mice,and the transcriptional levels of CCL2,CCL3 and MMP-12 were low(P all<0.01).In wild-type mice,total cell number,macrophage count and neutrophil count in alveolar lavage fluid of BALB/c mice treated with IL-17A monoclonal antibody were lower than those of control mice.The transcription levels of CCL2,CCL3 and MMP-12 were lower than those of control mice(P all<0.01).CS induces compensatory expression of IL-17A in macrophages,neutrophils and NK/NKT cells of NOD SCID mice.Conclusion The absence/reduction of IL-17A in CS-induced mice model can attenuate pulmonary inflammatory response,and IL-17A can be secreted by macrophages and neutrophils other than T cells in mice lungs.