模拟微重力下小鼠巨噬细胞感染空间诱变大肠埃希菌对炎性因子分泌和核因子κB表达的影响  

Effects of infection with space mutagenic E.coli on the inflammatory cytokines secretion and NF-κB expression in RAW264.7 macrophages under simulated microgravity

作　　者：裴雪枫 张晓毅 姚静[3] 刘蓉[3] 刘长庭[4] 王静宇[5] 袁明[5] 

机构地区：[1]锦州医科大学研究生学院火箭军总医院基地,辽宁锦州121001 [2]火箭军总医院泌尿外科,北京100088 [3]石河子大学医学院,新疆石河子832000 [4]解放军总医院南楼呼吸科,北京100853 [5]中国航天员科研训练中心航天医学基础与应用国家重点实验室,北京100094

出　　处：《解放军医学院学报》2017年第5期459-462,477,共5页Academic Journal of Chinese PLA Medical School

基　　金：国家重点基础研究发展计划课题(2014CB744404)~~

摘　　要：目的研究模拟微重力状态下小鼠巨噬细胞感染空间诱变大肠埃希菌(T1-13)后炎症反应及NF-κB表达的变化。方法巨噬细胞随机分为4组:对照组(Con)、对照染菌组(Con+T1-13)、模拟微重力组(SM)、模拟微重力染菌组(SM+T1-13)。细胞回转器模拟微重力效应,RT-q PCR检测细胞内炎性因子及NF-κB p65 m RNA表达;液相悬浮芯片多因子检测平台技术及硝酸还原酶法检测细胞培养液中炎性因子和总一氧化氮浓度变化;Western blot检测细胞中NF-κB信号通路相关蛋白表达。结果染菌后,对照染菌和模拟微重力染菌组细胞分泌的炎性因子浓度、细胞内炎性因子及NF-κB p65m RNA表达均显著升高;与对照组相比,模拟微重力组细胞培养液中TNF-α、IL-1β、IL-6分别升高168%、77.6%和187%;与对照染菌组相比,模拟微重力染菌组分泌炎性因子TNF-α、IL-1β、IL-6分别升高1.06%、6.78%、27.1%。巨噬细胞感染空间诱变大肠埃希菌后NF-κB p65 m RNA表达量显著升高,但是蛋白表达量显著下降,细胞内NF-κB激酶抑制剂β(IKKβ)、核因子κB激酶抑制剂α(IKKα)、磷酸化NF-κB p65蛋白表达量也显著下降。结论模拟微重力可诱导巨噬细胞炎性因子分泌升高,空间诱变大肠埃希菌刺激后这种升高反应受到抑制,可能与NF-κB蛋白表达下调有关。Objective To explore the effects of infection with space mutagenic E.coli on secretion of inflammatory cytokines and NF-κB expression in RAW264.7 macrophages under simulated microgravity.Methods RAW264.7 macrophages were randomly divided into four groups as Control group（Con）,Conrol＋T1-13 group（Con＋T1-13）,Simulated microgravity group（SM） and Simulated microgravity ＋T1-13 group（SM＋T1-13）,in which T1-13 is space mutagenic E.coli.A clinostat was used to simulate microgravity on cultured macrophages,then RAW264.7 macrophages were co-cultured with T1-13 E.coli in Con＋T1-13 and SM＋T1-13 groups for 6 hours.The m RNA expression of TNF-α,IL-1β,IL-6 and NF-κB p65 in macrophages were determined by RT-q PCR,the cytokines concentrations of TNF-α,IL-1β and IL-6 in culture medium were determined by multiplex immunoassay of Luminex x MAP platform,and the total Nitric Oxide Assay Kit was used to detect total NO in cell culture medium.Western Blot was used to detect the protein expression of NF-κB pathway related molecules.Results Compared with Con group,TNF-α,IL-1β and IL-6 concentration in SM group increased by 168%,77.6% and 187%,respectively.While compared with Con＋T1-13 group,TNF-α,IL-1β and IL-6 concentration in SM＋T1-13 group only increased by 1.06%,6.78% and 27.1%,respectively.After infected by space mutagenic E.coli,the NF-κB p65 m RNA expression in RAW264.7 increased significantly.Interestingly,the protein expression of IKKβ,IKKα and Phospho-NF-κB p65 and NF-κB p65 decreased significantly.Conclusion Simulated microgravity induces the increased cytokine secretion in macrophages,which can be inhibited by infection of space mutagenic E.coli.Such inhibition effects might be related with decreased protein expression of NF-κB.

关 键 词：空间诱变大肠埃希菌 模拟微重力 巨噬细胞 核因子ΚB 炎性因子 

分 类 号：R85[医药卫生—航空、航天与航海医学]