调节性T细胞对脓毒症中T淋巴细胞凋亡的影响  

The potential effect of regulatory t cells on promoting apoptosis of T lymphocytes in sepsis

作　　者：栾樱译[1] 秦庆华[1] 吕奔[2] 董宁[1] 于燕[1] 姚咏明[1] 

机构地区：[1]解放军总医院第一附属医院创伤研究中心,北京100048 [2]中南大学脓毒症转化医学湖南省重点实验室

出　　处：《临床急诊杂志》2016年第8期585-589,共5页Journal of Clinical Emergency

基　　金：国家自然科学基金(No:81130035;813720541;81272090;81401592);国家重点基础研究发展计划项目(No:2012CB518102)

摘　　要：目的:探讨CD4^+CD25^+调节性T细胞(CD4^+CD25^+Treg)对脓毒症过程中效应性T细胞(Teff)凋亡的影响。方法:应用SPF级健康雄性BALB/C小鼠制备盲肠结扎穿孔术(CLP)脓毒症模型。小鼠随机分为正常对照组、假伤组、CLP组、CLP+PC61(Treg特异性抑制剂)组、CLP+HRPN(PC61的同型对照蛋白IgG)组。免疫磁珠法分选小鼠脾脏CD4^+CD25^+Treg。采用流式细胞术观察T淋巴细胞毒性相关抗原(CTLA)-4及转录因子叉头翼状螺旋转录因子(Foxp3)表达以及CD25比例的变化,并检测各组小鼠CD4+CD25-T细胞的凋亡率。ELISA法检测外周血中白细胞介素(IL)-10、IL-2、IL-4和干扰素(IFN)-γ的含量变化。结果:CLP+PC61组术后48h小鼠生存率(90%)较CLP组(60%)明显升高;CLP+PC61组CD4^+CD25^+Treg中Foxp3及CTLA-4表达恢复至正常对照组水平,与CLP组差异有统计学意义(P<0.05),并且CLP+PC61组CD25表达水平最低。CLP+PC61组IL-2、IFN-γ水平显著升高,与CLP组差异明显(P<0.05),但IL-4、IL-10水平较CLP组出现不同程度地下降(P<0.05)。此外,CLP+PC61组效应性T细胞凋亡率明显低于CLP组(P<0.05)。结论:拮抗小鼠体内调节性T细胞活性可有效地减轻脓毒症状态下效应性T细胞凋亡,提高小鼠生存率。Objective：To investigate the potential effect of regulatory T cell（Treg）on promoting the apoptosis of T lymphocytes in mice with severe sepsis.Method：In this study,sepsis model was induced by cecal ligation and puncture surgery（CLP）.Male BALB/C mice used in the present experiment were randomly divided into 5groups as follows：normal control group,sham group,CLP group,CLP with PC61（the specific antagonist for Treg）treatment group,and CLP with HRPN（rat immunoglobulin G1）treatment group.Splenic CD4^＋CD25^＋Tregs and CD4^＋CD25^-T cells were isolated.Phenotypes,including cytotoxic T-lymphocyte-associated antigen 4（CTLA-4）,forkhead/winged helix transcription factor p3（Foxp3）and CD25,as well as the rate of apoptotic CD4^＋CD25^-T cell were analyzed by flow cytometry.Cytokines including interleukin（IL）-10,IL-2,IL-4and interferon（IFN）-γwere analyzed with ELISA.Result：The survival rate in the CLP＋PC61group（90%）at 48 hafter sepsis was significantly higher than that in the CLP group（60%）.The expressions of Foxp3 and CTLA-4were returned to the normal level in the CLP＋PC61group,and there was a significant difference between those of the CLP group and the CLP＋PC61group（P〈0.05）.Levels of IL-2and IFN-γwere markedly increased,while IL-4and IL-10 levels were significantly decreased in the CLP＋PC61group compared with those in the CLP group（P〈0.05）.In addition,the apoptosis rate of effector T cells in the CLP＋PC61group was markedly lower than that in the CLP group（P〈0.05）.Conclusion：The results suggested that inhibition of the activity of Tregs in mice could significantly decrease the apoptosis of effector T cells in sepsis and improve the survival rate of mice.

关 键 词：调节性T细胞 效应性T细胞 叉头翼状螺旋转录因子 T淋巴细胞毒性相关抗原4 凋亡 

分 类 号：R631[医药卫生—外科学]