牛磺酸影响人外周血T淋巴细胞增殖及分化功能的体外研究  

Effect of taurine on the proliferation and differentiation of human peripheral blood T lymphocytes in vitro

作　　者：孔星星 蓝利 袁江浪 陈成英 王新航 刘彧冰 李菡 陆彩玲 李习艺[3] 唐深[1,2] Kong Xingxing;Lan Li;Yuan Jianglang;Chen Chengying;Wang Xinhang;Liu Yubing;Li Han;Lu Cailing;Li Xiyi;Tang Shen(School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021,China;Key Laboratory of Basic Research on Regional Diseases,Education Department of Guangxi Zhuang Autonomous Region,Guangxi Medical University,Nanning 530021,China;School of Public Health,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学基础医学院,南宁530021 [2]广西高校区域性疾病基础研究重点实验室,南宁530021 [3]广西医科大学公共卫生学院,南宁530021

出　　处：《广西医科大学学报》2023年第5期742-748,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金项目(No.82260320,No.82160612);广西自然科学基金项目(No.2021GXNSFAA196019)。

摘　　要：目的:研究牛磺酸(Tau)对人外周血T淋巴细胞体外活化增殖及分化的影响及其作用机制。方法:无菌分离健康志愿者外周血淋巴细胞,建立植物凝集素(PHA)刺激T细胞体外活化增殖模型。实验设置对照组(control组)、PHA刺激组(PHA5μg/mL组)及Tau处理组(PHA+Tau 80 mmol/L组、PHA+Tau 160 mmol/L组)。细胞计数试剂盒(CCK-8)法检测T淋巴细胞增殖率;瑞—吉染色法观察细胞形态及计算转化率;实时荧光定量PCR检测T细胞增殖标志物Ki67,Th1转录因子T-bet及细胞因子干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α),Th2转录因子GATA-3及白介素-4/6(IL-4/6),凋亡相关因子Fas、FasL基因表达水平;活性氧(ROS)试剂盒检测细胞内ROS水平。结果:与control组相比,PHA 5μg/mL组T淋巴细胞转化率、增殖率、增殖标志物Ki67、Th1相关因子T-bet、IFN-γ、TNF-α、凋亡相关因子Fas基因表达及胞内ROS水平升高(P<0.05),凋亡相关因子FasL基因,Th2相关因子GATA-3、IL-6基因表达水平降低(P<0.05)。与PHA 5μg/mL组相比,PHA+Tau 80 mmol/L组与PHA+Tau 160 mmol/L组T淋巴细胞转化率、增殖率、Ki67基因表达及胞内ROS水平均下降(P<0.05),Th2相关因子IL-4、IL-6基因表达水平均升高(P<0.05),Th1相关因子IFN-γ、FasL基因表达水平差异均无统计学意义(P>0.05),PHA+Tau 80 mmol/L组Th1相关因子T-bet、TNF-α、Th2相关因子GATA-3及Fas基因表达水平差异无统计学意义(P>0.05),PHA+Tau 160 mmol/L组Th1相关因子T-bet、TNF-α、Th2相关因子GATA-3及Fas基因表达水平升高(P<0.05)。结论:Tau可能通过下调Ki67表达和增强Fas-AICD途径抑制T细胞增殖转化,并可能通过降低胞内ROS含量,提高Th1/Th2活化水平,调节Th1/Th2平衡偏向Th1分化,发挥调节T淋巴细胞活化增殖及分化的作用。Objective:To study the effect and mechanism of taurine(Tau)on the activation,proliferation,and differentiation of human peripheral blood T lymphocytes in vitro.Methods:Peripheral blood lymphocytes were aseptically isolated from healthy volunteers to establish the model of activation and proliferation of T cells stimulated by phytohaemagglutinin(PHA)in vitro.The control group,PHA stimulation group(PHA 5μg/mL group),and Tau treatment group(PHA+Tau 80 mmol/L group,PHA+Tau 160 mmol/L group)were set up.Cell counting kit-8(CCK-8)assay detected the T lymphocyte proliferation rate.The cell morphology was observed withWright-Giemsa staining and the conversion rate was calculated.T lymphocyte proliferation marker Ki67,Th1 transcription factor T-bet and cytokines interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),Th2 transcription factors GATA-3 and interleukin-4/6(IL-4/6)and the expressions of apoptosis-related factors Fas and FasL gene were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Reactive ox-ygen species(ROS)kits were used to detect the level of intracellular ROS.Results:Compared with the control group,T lymphocyte conversion rate,proliferation rate,proliferation marker Ki67,Th1-related factors T-bet,IFN-γ,TNF-α,apoptosis-related factor Fas gene expression and intracellular ROS level of PHA 5μg/mL group increased(P<0.05),while apoptosis-related factor FasL gene,Th2-related factors GATA-3 and IL-6 gene expression levels decreased(P<0.05).Compared with the PHA 5μg/mL group,T lymphocyte conversion rate,proliferation rate,Ki67 gene expression and intracellular ROS level in PHA+Tau 80 mmol/L group and PHA+Tau 160 mmol/L group decreased(P<0.05).The gene expression levels of Th2-related factors IL-4 and IL-6 increased(P<0.05).There were no significant differences in the gene expression levels of Th1-related factors IFN-γand FasL(P>0.05).There were no significant differences in the gene expression levels of Th1-related factors T-bet and TNF-α,and Th2-related factors GATA-3 and

关 键 词：牛磺酸 T淋巴细胞增殖 TH1/TH2分化 免疫调节 

分 类 号：R392[医药卫生—免疫学]