低剂量RT对小鼠肺泡巨噬细胞TLR4信号通路的影响  

Effect of low dose RT on TLR4 signaling pathway in mice alveolar maceophages

作　　者：王燕 柴绍明 黄燕霞[2] 孙成彪 边红 许娜[2,4] 刘文森 WANG Yan1 , CHAI Shao-ming2,3 , HUANG Yan-xia2 , SUN Cheng-biao2 , BIAN Hong1 , XU Na2,4 , LIU Wen-sen1,2(1. College of Life Science ,Jilin Agricultural University ,Changchun 130118,China ; 2. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130122, China ; 3. The Hospital of Infectious Diseases, Changchun 130041, China ; 4. Jilin Medical University ,Jilin City ,Jilin 132013 ,Chin)

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]军事医学科学院军事兽医研究所、吉林省人兽共患病防控重点实验室,吉林长春130122 [3]长春市传染病医院,吉林长春130000 [4]吉林医药学院,吉林吉林132013

出　　处：《中国兽医学报》2018年第5期986-990,997,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31372487)

摘　　要：取SPF级昆明小鼠进行RT雾化攻毒。攻毒后每隔一段时间通过BCA法测定小鼠支气管灌洗液BALF上清中总蛋白浓度，ELISA法测定小鼠BALF中致炎因子TNF-α、IL-6、IL-1β浓度，利用RT—PCR检测小鼠肺泡巨噬细胞TLR4及其信号通路mRNA的表达情况。结果显示：小鼠吸入1／150 LD50剂量雾化RT后，BALF上清总蛋白浓度增加，IL-1β、TNF-α浓度12h时达到最高值，IL-6浓度48h达到最高值。攻毒后12，24h时TLR4表达高于空白组，差异显著（0．01〈P〈0．05）；12h时MyD88表达高于空白组，差异极显著（P〈0．01）。12h时IRAK-1、IRAK-4、TRAF6表达高于空白组，差异显著（0．01〈P〈0．05）。结果表明：小鼠吸入1／150LD50剂量雾化RT，肺部发生炎症且肺泡组织通透性增加；低浓度蓖麻毒素攻毒后可刺激肺泡巨噬细胞TLR4及其信号转导通路下游MyD88、TRAF6、IRAK-1、IRAK-4等相关因子mRNA的表达，主要通过MyD88途径引起肺部炎症反应。To investigate changes of inflammatory cytokines in BALF supernatant and expression level of TLR4 and its downstream signaling pathway in pulmonary alveolar macrophages after sublethal dose aerosol ricin exposure. After aerosol ricin exposure,we took BCA method to determine the concentration of total protein in BALF supernatant, ELISA method to determine the concentration of inflammatory cytokines TNF-α,IL-6 ,IL-1β in BALF supernatant at each time points. We took RT-PCR method to determine the expression level of TLR4 and its downstream signaling pathway in pulmonary alveolar macrophages at each time points. The results showed that after aerosol ricin exposure at a dose of 1/150 LD50 ,the concentration of total protein was increased,the concentration of TNF-α and IL-1β reached a peak in 12 h after exposure,IL-6 reached the peak in 48 h. The expression of TLR4,MyD88 ,TRAF6 ,IRAK-1 and IRAK-4 was the highest at 12 h compared with other time points, showed a tendency of increasing first and then decreased indicating that after aerosol ricin exposure at a dose of 1/150 LD50 ,ricin trigger the pulmonary inflammation and increased permeability of alveolar tissue low dose of ricin exposure could mediate the TLR4 signaling pathway which is dependent on MyD88.

关 键 词：雾化蓖麻毒素 BALF 炎症 TLR4 

分 类 号：S852[农业科学—基础兽医学]