结核分枝杆菌H37Ra感染的THP-1细胞TGF-β1、Smad2、Smad3、TNF-α、IL-6 mRNA表达变化  

Expression changes of TGF-β1,Smad2,Smad3,TNF-α,and IL-6 mRNA in THP-1 cells infected with mycobacterium tuberculosis H37Ra

作　　者：王立楠 李艳宁 刘建[2] 屈昱良[2,3] 郭乐 杨宗强 牛宁奎[1] WANG Linan;LI Yanning;LIU Jian;QU Yuliang;GUO Le;YANG Zongqiang;NIU Ningkui(Department of Spinal Orthopaedics,General Hospital of Ningxia Medical University,Yinchuan 750004,China;不详)

机构地区：[1]宁夏医科大学总医院骨科,银川750004 [2]宁夏医科大学 [3]宁夏医科大学临床医学院宁夏临床病原微生物重点实验室

出　　处：《山东医药》2022年第3期44-49,共6页Shandong Medical Journal

基　　金：国家自然科学基金项目(81501903,81860395);宁夏自然科学基金项目(2020AAC03391);宁夏医科大学国内一流建设学科临床医学资助项目(NXYLXK2017A05);自治区卫生健康系统科研课题项目(2019-NW-011)。

摘　　要：目的观察结核分枝杆菌H37Ra在不同感染复数及不同感染时间感染的THP-1细胞内TGF-β1/Smad2、3信号通路活化以及炎症因子TNF-αmRNA、IL-6 mRNA的表达变化,明确TGF-β1/Smads经典信号通路的激活程度是否与结核分枝杆菌感染的细菌量有关以及在结核分枝杆菌感染后的哪个时间阶段发挥主要作用。方法培养THP-1巨噬细胞及结核分枝杆菌H37Ra,建立H37Ra在相同时间点(感染24 h)不同感染复数(MOI=0、1、10、100)和在MOI=10,不同感染时间点(T=0 h、6 h、12 h、24 h)感染THP-1细胞的感染模型;用qRT-PCR检测不同感染复数及不同感染时间THP-1细胞中TGF-β1 mRNA、Smad2 mRNA、Smad3 mRNA、TNF-αmRNA、IL-6 mRNA。结果H37Ra感染24 h时,与空白对照比较,当MOI=1、10、100时,TGF-β1 mRNA的相对表达量先升高后降低,在MOI=10时表达最高(P<0.05);Smad2 mRNA的相对表达量降低(P<0.05);Smad3 mRNA相对表达量升高(P<0.05);TNF-αmRNA和IL-6 mRNA表达变化趋势基本一致,在MOI=1、10时升高不明显(P>0.05),MOI=100时升高(P<0.05)。H37Ra MOI=10时,随感染时间(T=6 h、12 h、24 h)延长,TGF-β1 mRNA的相对表达量呈现升高趋势(P均<0.05);Smad2 mRNA的相对表达量呈降低趋势(P<0.05);Smad3 mRNA相对表达量呈先升高后降低趋势,在T=6 h最高(P<0.05);TNF-αmRNA和IL-6 mRNA表达变化趋势基本一致,呈先升高后降低趋势,在T=12 h时最高(P<0.05)。结论相同感染时间,随着MOI值增加,结核分枝杆菌H37Ra感染的THP-1巨噬细胞中TGF-β1 mRNA表达先上调后降低,Smad2 mRNA表达下调,Smad3 mRNA表达上调,TNF-αmRNA、IL-6 mRNA相对表达量在MOI=100时升高;MOI=10时,随着感染时间延长,结核分枝杆菌H37Ra感染的THP-1巨噬细胞中TGF-β1 mRNA相对表达量升高,Smad2 mRNA表达下调,Smad3 mRNA表达先升高,6 h后降低,TNF-αmRNA和IL-6 mRNA表达早期上调,12 h后降低。结核分枝杆菌感染的细菌量与TGF-β1/Smads经典信号通路的激活程度有关,结核分枝杆菌感染后的Objective pathway and the expression of inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in THP-1 cells infected with different multiple of infection(MOI)and different infection time of mycobacterium tuberculosis(MTB)H37 Ra,and to determine whether the activation of TGF-β1/Smads classical signaling pathway is related to the amount of bacteria infected by MTB and at which time stage it plays a major role after MTB infection.Methods phages and MTB H37 Ra were cultured to establish the infection models of THP-1 cells infected by H37 Ra in different MOI(0,1,10,and 100)and at the same time of 24 h and at different infection time(0,6,12,24)and in the same MOI(10). The mRNA expression levels of TGF-β1,Smad2,Smad3 and TNF-α and IL-6 in cells under different infection conditions were detected by quantitative real-time PCR(qRT-PCR).Results=0,T=0 h),at the same time of infection(T=24 h),when MOI=1,10,and 100,the expression level of TGF-β1 first increased and then decreased,and the expression was the highest when the infection time was 24 h and MOI=10(P<0. 05);the expression level of Smad2 was down-regulated and the expression of Smad3 was up-regulated(both P<0. 05);the expression of TNF-α and IL-6 showed the same trend,and the up-regulation was not obvious when MOI=1 and 10(all P>0. 05)and the expression was significantly up-regulated when MOI=100(P<0. 05). At MOI=10 and different infection time points(T=6,12,and 24 h),TGF-β1 expression showed a gradually up-regulated trend over the infection time(P<0. 05);the expression level of Smad2 was down-regulated significantly;the expression of Smad3 first increased and then decreased over the infection time,and the expression was the highest when the infection T=12 h and MOI=10(all P<0. 05). The expression levels of TNF-α and IL-6 showed the same trend,the expression of TNF-α and IL-6 were firstly upregulated and then down-regulated over the infection time,and the expression was the highest when the infection T=12 h and MOI=10(all P<0. 05).Conclus

关 键 词：结核分枝杆菌 巨噬细胞 转化生长因子β1/Smads信号通路 肿瘤坏死因子 白细胞介素6 

分 类 号：R378.9[医药卫生—病原生物学]