Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunc...Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.展开更多
目的探讨柴胡软坚消瘿颗粒对肝郁脾虚型桥本甲状腺炎大鼠辅助性T细胞1(helper T cell 1,Th1)/辅助性T细胞2(helper T cell 2,Th2)、辅助性T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)平衡的影响。方法随机数...目的探讨柴胡软坚消瘿颗粒对肝郁脾虚型桥本甲状腺炎大鼠辅助性T细胞1(helper T cell 1,Th1)/辅助性T细胞2(helper T cell 2,Th2)、辅助性T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)平衡的影响。方法随机数字表法将48只大鼠分为正常组(12只)和造模组(36只),采用甲状腺球蛋白注射结合高碘饮水+慢性束缚+游泳过劳+饮食失节制备肝郁脾虚桥本甲状腺炎大鼠模型,共33只大鼠造模成功,将造模成功大鼠随机分为模型组、金水宝片[0.45 g/(kg·d)]和柴胡软坚消瘿颗粒组[11.43 g/(kg·d)],每组11只,连续给药8周。观察大鼠一般情况,实验前后记录大鼠体重、摄食量、粪便含水量,实验结束后旷场实验分析大鼠行为;末次给药后12小时,取血,流式细胞术检测Th1、Th2、Th17、Treg细胞百分比;酶联免疫吸附法检测大鼠血清甲状腺球蛋白抗体(thyroglobulin antibody,TGAb)、甲状腺过氧化酶抗体(thyroidperoxidase antibody,TPOAb)、游离三碘甲状腺原氨酸(free triiodothyronine,FT3)、游离甲状腺素(freethyroxine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)、γ-干扰素(γ-interferon,IFN-γ)、白细胞介素10(interleukin 10,IL-10)、白细胞介素17(interleukin 17,IL-17)、白细胞介素35(interleukin35,IL-35)的水平;取血后分离大鼠甲状腺组织,苏木素—伊红染色法检测大鼠甲状腺组织病理形态;蛋白免疫印迹法检测大鼠甲状腺组织T盒子转录因子(T-box transcription factor,T-bet)、GATA结合蛋白3(GATA binding protein 3,GATA3)、叉头盒蛋白p3(forkhead box protein p3,Foxp3)、维A酸相关孤独受体γt(retinoid related orphan receptorγt,RORγt)的表达。结果与正常组相比,模型组大鼠甲状腺损伤严重,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著升高(P<0.01),TSH、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著降低(P<0.01);与模型组相比,金水宝片组、柴胡软坚消瘿颗粒组大鼠甲状腺损伤减轻,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著降低(P<0.01),TSH水平、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著升高(P<0.01)。结论柴胡软坚消瘿颗粒对肝郁脾虚桥本甲状腺炎大鼠治疗有一定的疗效,可能与调节Th1/Th2及Th17/Treg细胞平衡有关。展开更多
基金This study was supported by the National Natural Science Foundation of China(No.30672178)National Basic Research Program of China(No.2005CB522602)the National Natural Science Outstanding Youth Foundation of China(No.30125020).
文摘Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.
文摘目的探讨柴胡软坚消瘿颗粒对肝郁脾虚型桥本甲状腺炎大鼠辅助性T细胞1(helper T cell 1,Th1)/辅助性T细胞2(helper T cell 2,Th2)、辅助性T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)平衡的影响。方法随机数字表法将48只大鼠分为正常组(12只)和造模组(36只),采用甲状腺球蛋白注射结合高碘饮水+慢性束缚+游泳过劳+饮食失节制备肝郁脾虚桥本甲状腺炎大鼠模型,共33只大鼠造模成功,将造模成功大鼠随机分为模型组、金水宝片[0.45 g/(kg·d)]和柴胡软坚消瘿颗粒组[11.43 g/(kg·d)],每组11只,连续给药8周。观察大鼠一般情况,实验前后记录大鼠体重、摄食量、粪便含水量,实验结束后旷场实验分析大鼠行为;末次给药后12小时,取血,流式细胞术检测Th1、Th2、Th17、Treg细胞百分比;酶联免疫吸附法检测大鼠血清甲状腺球蛋白抗体(thyroglobulin antibody,TGAb)、甲状腺过氧化酶抗体(thyroidperoxidase antibody,TPOAb)、游离三碘甲状腺原氨酸(free triiodothyronine,FT3)、游离甲状腺素(freethyroxine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)、γ-干扰素(γ-interferon,IFN-γ)、白细胞介素10(interleukin 10,IL-10)、白细胞介素17(interleukin 17,IL-17)、白细胞介素35(interleukin35,IL-35)的水平;取血后分离大鼠甲状腺组织,苏木素—伊红染色法检测大鼠甲状腺组织病理形态;蛋白免疫印迹法检测大鼠甲状腺组织T盒子转录因子(T-box transcription factor,T-bet)、GATA结合蛋白3(GATA binding protein 3,GATA3)、叉头盒蛋白p3(forkhead box protein p3,Foxp3)、维A酸相关孤独受体γt(retinoid related orphan receptorγt,RORγt)的表达。结果与正常组相比,模型组大鼠甲状腺损伤严重,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著升高(P<0.01),TSH、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著降低(P<0.01);与模型组相比,金水宝片组、柴胡软坚消瘿颗粒组大鼠甲状腺损伤减轻,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著降低(P<0.01),TSH水平、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著升高(P<0.01)。结论柴胡软坚消瘿颗粒对肝郁脾虚桥本甲状腺炎大鼠治疗有一定的疗效,可能与调节Th1/Th2及Th17/Treg细胞平衡有关。
