文章摘要
高 梅,孙 杰,金文杰,钱燕宁.氯胺酮对离体人辅助T细胞分化的影响[J].南京医科大学学报,2011,(12):1741~1747
氯胺酮对离体人辅助T细胞分化的影响
Effect of ketamine on the differentiation of human T helper cells in vitro
投稿时间:2011-08-29  
DOI:10.7655
中文关键词: 氯胺酮  辅助T细胞  细胞因子  电泳迁移率实验
英文关键词: ketamine  T helper cell  cytokine  EMSA
基金项目:国家自然科学基金资助(30801068)
作者单位
高 梅 南京医科大学第一附属医院麻醉科,江苏 南京 210029 
孙 杰 南京医科大学第一附属医院麻醉科,江苏 南京 210029 
金文杰 南京医科大学第一附属医院麻醉科,江苏 南京 210029 
钱燕宁 南京医科大学第一附属医院麻醉科,江苏 南京 210029 
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中文摘要:
      目的:探讨氯胺酮对离体人辅助性T(Th)细胞分化和细胞内转录因子T-bet及GATA3活性的影响?方法:10例男性健康志愿者(20~45岁,BMI 15~25 kg/m2),分别采集空腹时外周静脉血20 ml,分离外周血单核细胞(PBMCs)?每份血样的PBMCs均随机加入佛波醇-十四酸酯-乙酸酯(PMA)25 ng/ml和离子霉素1 ?滋g/ml以及各种浓度(0?50?100?200 ?滋mol/L)的氯胺酮孵育4 h?然后测量Th细胞亚群百分数?上清液中的干扰素-γ(IFN-γ)和白介素-4(IL-4)的水平以及细胞内转录因子T-bet和GATA3的DNA结合活性?结果:在PMA和离子霉素刺激下Th1和Th2细胞计数?IFN-γ和IL-4水平均显著增加;在PMA和离子霉素刺激下,氯胺酮剂量依赖性降低Th1和Th2细胞计数?IFN-γ和IL-4水平,而增加Th1?Th2细胞计数的比值(Th1/Th2)和IFN-γ?IL-4的比值(IFN-γ/IL-4);同样经刺激后,细胞内T-bet和GATA3的活性显著增加;氯胺酮剂量依赖性降低T-bet和GATA3的活性,而增加T-bet与GATA3活性的比值(T-bet/GATA3)?结论:在PMA和离子霉素的刺激下,氯胺酮能够抑制Th细胞的分化,从而抑制各细胞因子的分泌,但Th1/Th2增加;氯胺酮增加Th1/Th2可能与调节增加T-bet/GATA3有关?
英文摘要:
      Objective:To investigate the effect of ketamine on the differentiation of human T helper(TH) cells and activity of T-bet and GATA3 in vitro. Methods: Ten male healthy volunteers (aged 20~45y,BMI 18~25 kg/m2) were enrolled in this study. Tweenty milliliter peripheral venous blood was collected from each volunteer and peripheral blood mononuclear cells(PBMC) were isolated and incubated in the presence of phorbol-myristate-acetate (PMA)(25 ng/ml) plus ionomycin (1 ?滋g/ml) and various concentrations of ketamine(0,50,100,200 ?滋mol/L). TH subsets,supernatant interferon γ (IFN-γ) and interleukin 4 (IL-4) levels,and the DNA binding activity of intracellular transcriptional factors T-bet and GATA3 were detected and analyzed after four hours culture. Results: TH1 and TH2 cell numbers,IFN-γ and IL-4 levels were significantly increased by PMA and ionomycin stimulation. Ketamine dose-dependently decreased TH1 and TH2 cell numbers,IFN-γ,and IL-4 levels,but increased the ratio of TH1/TH2 and IFN-γ/IL-4 in the presence of PMA and ionomycin. T-bet and GATA3 activities in PBMC were significantly increased by PMA and ionomycin stimulation. Ketamine dose-dependently decreased T-bet and GATA3 activity,but increased the ratio of T-bet/GATA3 activity in the presence of PMA and ionomycin. Conclusion: Ketamine can suppress TH cell differentiation and subsequent cytokines. But it increase TH1/TH2 ratio after PMA and ionomycin stimulation,which may be related to its regulation on T-bet and GATA3 activity.
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