雾化吸入脂多糖导致的大鼠急性肺炎模型不同标本中炎性因子比较研究  

Comparative research of inflammatory factors in different specimens from a rat model of aspiration pneumonia induced by lipopolysaccharide

作　　者：宋玲 高云航 李晗 陈腾飞 侯红平[1] 叶祖光[1] 张广平[1] SONG Ling;GAO Yunhang;LI Han;CHEN Tengfei;HOU Hongping;YE Zuguang;ZHANG Guangping(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 10070,China)

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国实验动物学报》2025年第1期89-98,共10页Acta Laboratorium Animalis Scientia Sinica

基　　金：中药研究所中药药理创新团队项目(CI2021B015);中国中医科学院创新工程项目(C12021A04615);国家自然科学基金青年项目(82204740)。

摘　　要：目的探究通过雾化吸入脂多糖(LPS)所诱发的大鼠急性肺炎模型中,肺泡灌洗液、肺组织以及血清中炎性因子的变化情况。方法通过连续3 d、每天雾化吸入15 min 4 mg/mL的LPS来诱导大鼠急性肺炎模型,并同步监测LPS的粒径分布及气溶胶浓度,苏木素-伊红(HE)染色观察大鼠肺病理学形态变化,显微镜观察肺泡灌洗液涂片白细胞及计数;超敏多因子电化学发光法测肺组织、血清及肺泡灌洗液干扰素-γ(IFN-γ)、白细胞介素(IL)-1β、IL-4、IL-5、IL-6、IL-10、IL-13、肿瘤坏死因子-α(TNF-α)、角质形成细胞衍生趋化因子(keratinocyte-derived chemokine/growth-regulated oncogene,KC/GRO)炎性因子。结果LPS诱导的肺炎模型组织病理学,肺可见局灶或弥漫、肺泡上皮坏死伴脱落、炎性细胞浸润。测定雾化LPS累积粒度分布达到10%、50%及90%对应的粒径(diameter of 10%,50%and 90%volume distribution,Dv(10),Dv(50)及Dv(90))分别为0.6974μm、3.387μm和8.836μm,气溶胶浓度为4.08 g/m^(3),计算大鼠的吸入剂量为47.10 mg/kg。在肺泡灌洗液中,模型组的中性粒细胞数量显著增加(P<0.01);同时模型组肺组织中的炎性因子IL-1β、IL-6、KC/GRO水平显著上升(P<0.01,P<0.05,P<0.01)。肺泡灌洗液中的炎性因子IL-1β、IL-6、TNF-α的含量也显著增高(P<0.05,P<0.001,P<0.001)。而血清中的炎性因子则未见明显变化。结论在雾化吸入LPS构建的急性肺炎模型中,IL-1β、IL-6、KC/GRO以及TNF-α等炎性因子在肺组织和肺泡灌洗液中均出现了显著变化,而血清中的炎性因子则未见明显变化。这表明该模型主要引发的是肺部局部的炎症反应。Objective To investigate the changes of inflammatory factors in bronchoalveolar lavage fluid(BALF),lung tissue,and serum of a rat pneumonia model induced by inhalation of lipopolysaccharides(LPS).Methods Three days after modeling by LPS 4 mg/mL inhalation,15 min/d,was conducted while monitoring the particle size distribution and aerosol concentration of LPS,the degree of inflammation in lung tissues of rats in each group was observed via HE staining,and neutrophils in BALF were counted by microscope.The contents of interferon gamma(IFN-γ),interleukin-1 beta(IL-1β),IL-4,IL-5,IL-6,IL-10,IL-13,tumor necrosis factor alpha(TNF-α),and KC/GRO in lung tissue,serum,and BALF were detected by Meso Scale Discovery.Results The lung histopathology of model rats displayed focal and diffuse alveolar epithelial necrosis with shedding and the aggregation and infiltration of inflammatory cells.The particle size distribution of atomized LPS was as follows,Dv(10)=0.6974μm,Dv(50)=3.387μm,Dv(90)=8.836μm.The aerosol concentration of LPS was 4.08 g/m^(3),and the calculated inhalation dose for rats was 47.10 mg/kg.The neutrophil count(P<0.01)and contents of IL-1β,IL-6,and TNF-α(P<0.05,P<0.001,P<0.001)in the BALF,and the contents of IL-1β,IL-6,and KC/GRO in lung tissue(P<0.01,P<0.05,P<0.01),of model rats were significantly increased.No biologically significant changes were observed in inflammatory factor levels in the serum.Conclusions In the acute pneumonia model induced by inhalation of LPS,significant changes in inflammatory factors such as IL-1β,IL-6,KC/GRO,and TNF-αwere observed in both lung tissue and bronchoalveolar lavage fluid(BALF),while no notable changes in these inflammatory factors were detected in serum.This indicates that the inflammation responses are primarily localized in the lungs.

关 键 词：急性肺炎模型 脂多糖 雾化吸入 炎性因子 白细胞分类计数 

分 类 号：Q95-33[生物学—动物学]