GATA-3和T—bet与吗啡抑制成人辅助性T淋巴细胞分化的关系  

Relationship between GATA-3 and T-bet and inhibition of differentiation of human T helper cells by morphine

作　　者：曹国平[1] 毛毛[2] 钱燕宁[2] 孙杰[2] 

机构地区：[1]江苏大学附属金坛医院麻醉科,213200 [2]南京医科大学第一附属医院麻醉科

出　　处：《中华麻醉学杂志》2012年第9期1088-1090,共3页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30801086）

摘　　要：目的评价核蛋白转录因子GATA-3和T-bet与吗啡抑制成人辅助性T淋巴细胞（Th细胞）分化的关系。方法健康志愿者10名，年龄20-50岁，体重50-70妇，清晨空腹采集外周静脉血样20ml，分离外周血单个核细胞（PBMCs）。采用随机配伍设计，将每份血样分离出的PBMCs随机分为5组（n=10）：常规培养组（C组）、刺激剂刺激分化组（P组）、吗啡50μg／ml组（M组）、吗啡50μg／ml＋纳洛酮50μg／ml组（MN组）、纳洛酮50μg／ml组（N组）。刺激剂孵育4h后分别收集细胞和上清液。应用流式细胞术检测Th1、Th2细胞亚群比例，计算二者比值（Th1／Th2）；用酶联免疫吸附法检测上清液中干扰素（IFN）-7和白细胞介素（IL）-4的浓度；使用凝胶电迁移技术检测淋巴细胞核蛋白转录因子GATA-3和％bet的活性。结果与P组比较，M组Thl、Th2细胞亚群比例、Th1／Th2、上清液IFN-γ和IL-4的浓度、GATA-3和T-bet活性降低，MN组Th1细胞亚群比例、Th1／Th2、上清液IFN-γ浓度降低（P〈0．05或0．01）；与M组比较，MN组Th1、Th2细胞亚群比例、Th1／Th2、上清液IFN-γ和IL-4的浓度、GATA-3和bet活性升高（P〈0．05或0．01）；N组与C组上述各指标比较差异无统计学意义（P〉0．05）。结论吗啡通过激活阿片受体抑制Th细胞分化，而这种抑制作用与抑制核蛋白转录因子GATA-3和T-bet的活性有关。Objective To evaluate the relationship between GATA-3 and T-bet and inhibition of the dif- ferentiation of human T helper cells by morphine. Methods Ten healthy volunteers, aged 20-50 yr, weighing 50- 70 kg, were enrolled in the study. Peripheral venous blood samples were taken in the early morning. Peripheral blood mononuclear cells （PBMCs） were isolated and assigned into 5 groups （ n = 10 each）. PBMCs were incubated routinely （group C）. PBMCs were incubated in the presence of PMA and ionomycin （group P）, morphine 50 μg/ml （group M）, morphine 50 μg/ml ＋ naloxone 50 μg/ml （group MN） or naloxone 50μg/ml （group N）, and were then stimulated with PMA and ionomycin for another 4 h. The percentage of Thl and Th2 ceils was detected by flow cytometry. The Thl/Th2 ratio was calculated. Interferon （IFN）-γ and intedeukin （IL）-4 concentrations in the supernatant were determined using ELISA. The activities of GATA-3 and T-bet were analyzed by EMSA. IRe- suits Compared with group P, the percentage of Thl and Th2 cells, Thl/Th2 ratio, IFN-γ and ]L-4 concentra- tions in the supernatant, and GATA-3 and T-bet activities were significantly decreased in group M, the percentage of Th1 cells, Th1/Th2 ratio, and IFN-γ concentration in the supernatant were significantly decreased in group MN （P 〈 0.05 or 0.01）. Compared with group M, the percentage of Thl and Th2 cells, Th1/Th2 ratio, IFN-γ and IL-4 concentrations in the supernatant, and GATA-3 and T-bet activities were significantly increased in group MN （P 〈 0.05 or 0.01 ）. There was no significant difference in the indexes mentioned above between groups N and C （ P 〉 0.05）. Conduslon Morphine inhibits the differentiation of human T helper cells by activating opioid recep- tors, which may be related to the inhibition of GATA-3 and T-bet activities.

关 键 词：吗啡 T淋巴细胞 辅助诱导 GATA3转录因子 T-bet转录因子 

分 类 号：R614[医药卫生—麻醉学]