TMB-8对小鼠T淋巴细胞体外活化、增殖和细胞周期的影响  

Effect of TMB-8 on the activation, proliferation and cell-cycle distribution of the mouse T lymphocytes in vitro

作　　者：叶雪仪[1] 曾耀英[1] 黄秀艳[1] 王通[1] 臧宁[1] 周建国[1] 林长乐[1] 

机构地区：[1]暨南大学组织移植与免疫实验中心,广东广州510632

出　　处：《中国病理生理杂志》2007年第5期1008-1012,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学重点基金资助项目(No30230350);广东省自然科学基金资助项目(No5300413)

摘　　要：目的:研究胞内钙离子释放阻断剂8-(N,N-二乙胺)辛基-3,4,5-三甲氧基苯甲酸酯(TMB-8)对刀豆蛋白A(ConA)介导的小鼠T淋巴细胞体外活化、增殖和细胞周期的影响。方法:以ConA作为T细胞活化、增殖的刺激剂,以不同浓度的TMB-8及与环孢菌素A(CsA)联合作用于该系统,用流式细胞术检测T细胞早期活化标志CD69分子的表达;以活体染料羧基荧光素乙酰乙酸(CFDA-SE)染色流式细胞术,分析TMB-8在ConA刺激下小鼠淋巴细胞的增殖相关指数(PI);以碘化丙啶染色分析细胞周期的分布情况。结果:ConA作用6h后,CD69+T细胞的比率为(74.88±1.88)%,TMB-8在终浓度10、20、40μmol/L下均抑制ConA介导的T细胞CD69表达(P<0.01),其中,40μmol/L的TMB-8为(52.55±1.54)%,达到最高抑制率。培养48h和72h,ConA刺激下的PI值分别为1.24±0.01和2.05±0.07,TMB-8从5μmol/L起均抑制ConA介导的淋巴细胞增殖(P<0.01),以40μmol/L的效果最为显著,PI值分别为1.01±0.01和1.10±0.01;10μmol/L的TMB-8与25μg/L的环孢菌素A(CsA)具有明显的协同抑制作用(P<0.01)。细胞周期分析显示,培养48h的TMB-8从10μmol/L起即显著抑制S期(P<0.01)。结论:TMB-8可明显抑制ConA介导下的T细胞早期活化及增殖,并具T细胞周期的S期阻滞作用。AIM： To study the effects of [ 8 - （diethylamino） octyl - 3,4, 5 - trimethoxybenzoate ] （ TMB - 8）, an intracellular Ca^2＋ antagonist, on the activation, proliferation and cell - cycle distribution of the mouse T lymphocytes stimulated by concanvalin A （Con A） in vitro. METHODS： After stimulated with Con A, T cells were treated with different concentrations of TMB -8 alone and its combination with cyclosporine A （CsA）. The expression of CD69, the early marker of CD^3＋ T cell activation, was measured by FACS. The proliferation - related index was determined by carboxyl fluorescin diacetate succinmidyl ester （ CFDA - SE） flow cytometry. The cell - cycle distribution was analyzed by propidium iodide staining. RESULTS： After 6 h culture, the activation rate of CD69^＋ T cell in Con A group was （74.88±1.88） %. 10, 20 and 40 p, mol/L of TMB - 8 inhibited the expression of CD69 （ P 〈 0. 01 ）, especially in 40 μmol/L （52.55% ± 1.54% ）. After 48 h and 72 h culture, the PI of Con A group was 1.24 ± 0.01, 2. 05 ± 0. 07, respectively. TMB - 8 with the concentration up to 5 μmol/L exerted a definite inhibitory effect on the proliferation with a maximal inhi- bition in 40 p, mol/L（P 〈 O. O1 ）. In the combination of 10 p, mol/L of TMB - 8 with 25 p,g/L of CsA, an evident synergistic effect was observed （P 〈 0. 01 ）. Moreover, the cell - cycle distribution analysis showed that after 48 h culture, the concentration of TMB - 8 over 10μmol/L showed an evident suppression in S phase. CONCLUSION ： TMB - 8 significantly inhibites the early steps of the Con A - induced T cell activation and proliferation, as well as the progression of T lymphocytes in S phase.

关 键 词：三甲氧基苯甲酸酯 T淋巴细胞 CD69 细胞增殖 细胞周期 流式细胞术 

分 类 号：R363[医药卫生—病理学]