肿瘤坏死因子受体1在肺炎支原体肺炎小鼠固有免疫应答中的调节机制  

Regulatory mechanism of tumor necrosis factor receptor 1 in the innate immune response of mice with Mycoplasma pneumoniae pneumonia

作　　者：曾妮 张节平 吴文娟 邢晓莉 黄少祥 ZENG Ni;ZHANG Jie-ping;WU Wen-juan;XING Xiao-li;HUANG Shao-xiang(Tianjin Fifth Central Hospital,Tianjin 300450,China)

机构地区：[1]天津第五中心医院呼吸内科,天津300450

出　　处：《中华医院感染学杂志》2023年第12期1777-1782,共6页Chinese Journal of Nosocomiology

基　　金：天津市卫生局科技基金资助项目(2019KZ019)。

摘　　要：目的 探讨肿瘤坏死因子受体1(TNFR1)在肺炎支原体(MP)肺炎小鼠固有免疫应答中的调节机制。方法 采用随机数字法将小鼠分为对照组、模型组、敲低TNFR1组、过表达TNFR1组,各40只,构建肺炎支原体肺炎小鼠模型,检测小鼠肺泡灌洗液(BALF)中炎性细胞表达,HE染色观察肺组织炎症浸润与纤维化损伤程度,实时定量聚合酶链反应(RT-qPCR)检测肺组织中TNFR1及炎性因子mRNA转录水平,蛋白质免疫印迹法(WB)检测蛋白相对表达水平;采用MP转染鼠MC3T3细胞,并用TNFR1阻断抗体孵育转染后细胞,检测细胞培养液及细胞中TNFR1及炎性因子mRNA转录水平和蛋白相对表达水平。结果 过表达TNFR1组小鼠BALF中炎性细胞计数高于对照组、模型组、敲低TNFR1组(P<0.05);过表达TNFR1组TNFR1、肿瘤坏死因子-α(TNF-α)的mRNA转录水平、蛋白相对表达水平高于对照组、模型组、敲低TNFR1组,白细胞介素-1β(IL-1β)、IL-4、IL-10 mRNA转录水平、蛋白相对表达水平高于对照组、敲低TNFR1组,趋近于模型组(P<0.05);MP转染组MC3T3细胞及细胞培养液中TNFR1的mRNA转录水平、蛋白相对表达水平高于正常组(P<0.05);与MP转染组相比,TNFR1阻断组的TNFR1、TNF-α、IL-1β、IL-4、IL-10的mRNA转录水平、蛋白相对表达水平低(P<0.05)。结论 TNFR1能调节肺炎支原体肺炎小鼠固有免疫,TNFR1在肺炎支原体侵入机体后的炎性细胞活化和炎性因子的释放过程中具有促进作用,推测可能为机体的一种自身免疫机制。OBJECTIVE To investigate the regulatory mechanism of TNFR1 in the innate immune response of mice with Mycoplasma pneumoniae pneumonia.METHODS Mice were divided into control group,model group,knockdown TNFR1 group and overexpression TNFR1 group using random number method,with 40 mice in each group.The mouse model of MPpneumonia was established,the expression of inflammatory cells in BALF was detected.The degree of inflammatory infiltration and fibrosis injury in lung tissue was observed by HE staining.The transcription level of TNFR1 and inflammatory factor mRNA in lung tissue was detected by RT-qPCR,the relative protein expression levels were detected by protein immunoblotting(WB);murine MC3T3 cells were transfected with MP,and the transfected cells were incubated with TNFR1 blocking antibody.The mRNA transcription and protein expression levels of TNFR1 and inflammatory factors in cell culture medium and cells were detected and analyzed.RESULTS The inflammatory cell count in BALF of mice in the overexpressed TNFR1 group was higher than that of the control group,the model group,and the knock-down TNFR1 group(P<0.05);the mRNA transcription level and protein relative expression level of TNFR1,TNF-α,in the overexpressed TNFR1 group were higher than that of the control group,the model group,and the knock-down TNFR1 group,and the mRNA transcription level and protein relative expression level of IL-1β,IL-4,and IL-10 were higher than that of the control group,and the knock-down TNFR1 group,the model group,which were close to the model group(P<0.05);the mRNA transcript levels and relative protein expression levels of TNFR1 in MC3T3 cells and cell cultures of the MP transfection group were higher than those of the normal group(P<0.05);The mRNA transcription levels and relative protein expression levels of TNFR1,TNF-α,IL-1β,IL-4,and IL-10 were lower in the TNFR1 blocking group compared with the MP transfection group(P<0.05).CONCLUSION TNFR1 modulated the innate immunity of mice with MP pneumonia.TNFR1 had a facilitative

关 键 词：肿瘤坏死因子受体1 肺炎支原体 支原体肺炎 小鼠 固有免疫 调节机制 

分 类 号：R563.1[医药卫生—呼吸系统]