TLR4在脓毒症中对调节性T细胞功能活性的影响  

The effects of TLR4 on regulating the function of Tregs in sepsis

作　　者：黄颖 曹超[1] 王军[1] 寿松涛[1] HUANG Ying;CAO Chao;WANG Jun;SHOU Song-tao(Department of Emergency Medicine,General Hospital Tianjin Medical University,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院急诊科,天津300052

出　　处：《天津医科大学学报》2018年第6期492-495,共4页Journal of Tianjin Medical University

基　　金：天津市应用基础与前沿技术研究计划项目(13JCYBJC37500);睿E(睿医)急诊医学研究专项基金项目(R2015026);天普研究基金项目(UF201315);天津医科大学总医院青年孵育基金项目(ZYYFY2015010)

摘　　要：目的:探讨脓毒症中TLR4(TLR4)对调节性T细胞功能及活性的影响,并探讨其作用机制。方法:SPF级C57/BL6雄性小鼠(wild type,WT)40只,C57BL/10ScNJNJU(TLR4^(-/-))小鼠40只,随机均分为4组,WTSham组,WTCLP组,TLR4^(-/-)Sham组和TLR4^(-/-)CLP。CLP组用盲肠结扎穿孔法制备脓毒症模型。造模后24 h,FCM检测脾脏CD4^+CD25^+Foxp3^+Treg细胞数量、凋亡率及Foxp3和TLR4表达情况, RT-PCR检测Foxp3^+mRNA和TLR4mRNA的表达,Western Blot检测Treg细胞内NF-κB、p-NF-κB蛋白表达情况。结果:(1)TLR4^(-/-)CLP组Treg细胞凋亡率较WTCLP组明显增多,而细胞数量显著降低(P<0.05)。(2)WTCLP组Treg中Foxp3mRNA及蛋白表达、TLR4mRNA及蛋白水平显著升高(P<0.05),而在TLR4^(-/-)CLP组中该表达较WTCLP组均显著降低(P<0.05)。(3)WTCLP组Treg细胞NF-κB、p-NF-κB的表达水平明显升高(P<0.05),而TLR4^(-/-)CLP组较WTCLP组明显降低(P<0.05)。结论:在脓毒症中TLR4可显著影响Treg的功能及其活性,这可能通过TLR4/NF-κB信号通路介导有关。Objective To study the effects of TLR4 on regulating the function of Tregs in sepsis. Methods：Fortyhealthy male C57/BL mice（wild type,WT）and forty healthy male TLR4 knockout mice（TLR4^-/-）were divided into Sham operation group（WT Sham group,n =20）, WTCLP group（n=20）, and TLR4^-/-Sham group （n=20）, TLR4^-/-CLP group（n =20） by random number table method. In CLP group,the mice were given cecal ligation puncture（CLP）, and in Sham group,the mice were given Sham-operation. Twenty-four hours after CLP,Tregnumber,Tregapoptsis rate, the expression of Foxp3 ,TLR4 at mRNA and protein level, the protein expression of NF-κB and p-NF-κB were determined. Results： （1）The apoptosis of Treg cells in TLR4^-/-CLP group was higher than that in WTCLP group（P〈0.05） .（2）The levels of Foxp3mRNA and Foxp3 protein, TLR4mRNA and TLR4 protein in TLR4^-/-CLP group were significantly lower than those in WTCLP group（P〈0.05） . （3） The expression of NF-κB and p-NF-κB in splenic Tregs was significantly increased in WT CLP mice in comparison to TLR4^-/-CLP mice（P〈0.05） .Conclusion： TLR4 in experimental sepsiscloselyaligns with the function of Tregs, and this effect may be related to TLR4/NF-κB signaling.

关 键 词：TOLL样受体4 调节性T细胞 脓毒症 信号通路 

分 类 号：R631[医药卫生—外科学]