摘要
目的探讨白细胞介素-17(IL-17)对脂多糖(LPS)致急性肺损伤(ALI)小鼠炎症免疫调节的影响。方法将36只SPF级C57BL/6小鼠按随机数字表法分为生理盐水对照组(NS组)、LPS致ALI模型组(LPS组,气管内滴入LPS 5 mg/kg),每组18只。分别于制模后2、6、24 h处死6只小鼠,收集外周血、肺组织和脾组织,采用苏木素-伊红(HE)染色,光镜下观察肺组织病理学改变并检测淋巴细胞、中性粒细胞、巨噬细胞在肺泡壁及气管壁的浸润水平;采用免疫组化法检测肺泡壁及气管壁IL-17蛋白表达,分析IL-17蛋白表达与肺泡壁及气管壁淋巴细胞、中性粒细胞、巨噬细胞浸润的相关性;采用酶联免疫吸附试验(ELISA)测定肺组织匀浆中IL-17含量;采用流式细胞仪检测外周血、肺组织、脾组织中CD4+IL-17+辅助性T细胞(Th17细胞)占CD4+?T细胞的比例。结果①光镜下观察,NS组制模后各时间点肺组织结构基本正常,无明显炎性细胞浸润;LPS组制模后肺组织水肿、炎症反应逐渐加重,各时间点肺损伤评分(分)明显高于NS组(2 h:4.47±1.42比1.10±0.55,6 h:7.93±2.14比1.23±0.50,24 h:12.67±2.67比1.20±0.61,均P<0.01)。②免疫组化结果显示,随时间延长,LPS组肺泡壁及气管壁IL-17蛋白表达逐渐增强,而NS组则呈阴性或弱阳性表达;定量分析显示,LPS组各时间点肺泡壁及气管壁IL-17蛋白免疫组化染色评分(分)均高于NS组(肺泡壁:2 h为2.70±1.40比0.90±0.37,6 h为5.10±1.76比1.17±0.59,24 h为9.67±1.32比1.10±0.45;气管壁:2 h为2.87±0.89比0.90±0.39,6 h为4.97±1.48比1.10±0.41,24 h为8.67±1.54比1.03±0.29,均P<0.05)。③相关性分析显示,肺泡壁、气管壁中IL-17蛋白表达与淋巴细胞、中性粒细胞、巨噬细胞数量均呈正相关(肺泡壁:r值分别为0.632、0.550、0.466,气管壁:r值分别为0.695、0.662、0.575,均P<0.01)。④LPS组制模后肺组织匀浆液中IL-17含量(μg/L)逐渐升高,各时间点明显高于NS组(2 h:1.37±0.14比1.01±0
Objective To explore the immunomodulatory effects of interleukin-17(IL-17)on acute lung injury(ALI)induced by lipopolysaccharide(LPS).Methods Thirty-six SPF-class C57BL/6 mice were divided into normal saline control group(NS group)and LPS-induced ALI model group(LPS group,LPS 5 mg/kg intratracheal drip)according to random number table method,with 18 mice in each group.Six mice were sacrificed at 2,6 and 24 hours after model reproduction,and peripheral blood,lung and spleen tissues were harvested.After staining with hematoxylin-eosin(HE),the pathological changes of lung tissue were observed under microscope and the infiltration level of lymphocytes,neutrophils and macrophages in the alveolar wall and tracheal wall were detected.Immunohistochemistry was used to detect the protein expression of IL-17 in alveolar wall and tracheal wall,and the correlation between IL-17 expression and lymphocytes,neutrophils and macrophages infiltration in alveolar wall and tracheal wall were analyzed.The level of IL-17 in lung tissue homogenate was determined by enzyme linked immunosorbent assay(ELISA).Flow cytometry was used to detect the proportion of CD4+IL-17+helper T cells(Th17 cells)in CD4+T cells in peripheral blood,lung tissue and spleen tissue.Results①Microscopy showed that the lung tissue structure of NS group was basically normal at each time after model reproduction,and there was no obvious inflammatory cell infiltration,while the lung tissue edema and inflammatory reaction were gradually aggravated in the LPS group,and the lung injury score was significantly higher than that in NS group at each time(2 hours:4.47±1.42 vs.1.10±0.55,6 hours:7.93±2.14 vs.1.23±0.50,24 hours:12.67±2.67 vs.1.20±0.61,all P<0.01).②Immunohistochemistry showed that the protein expression of IL-17 in alveolar wall and tracheal wall of LPS group increased gradually with time,while that in NS group was negative or weak positive.Quantitative analysis showed that the immunohistochemical staining score of IL-17 protein in alveolar wall and trac
作者
秦翌佳
老启芳
黄冰
黎阳
覃韬
黄英明
Qin Yijia;Lao Qifang;Huang Bing;Li Yang;Qin Tao;Huang Yingming(Department of Critical Care Medicine,Guangxi Medical University Cancer Hospital,Nanning 530021,Guangxi Zhuang Autonomous Region,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2019年第8期983-988,共6页
Chinese Critical Care Medicine
基金
广西自然科学基金(2015GXNSFBA139146)
广西壮族自治区卫生健康委员会自筹经费课题(Z2014243).
