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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1μg/ml以及各种浓度(0、50、100、200μmol/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。
AIM: To investigate the effects of ketamine on the differentiation of human T helper (T) cells and the expression of T-bet and GATA3 in vitro. Methods: Ten healthy male volunteers (20-45 years old, BMI 15-25 kg / m2) were enrolled in this study. Peripheral venous blood (20 ml) was collected and peripheral blood mononuclear cells (PBMCs) were isolated. PBMCs from each blood sample were randomly assigned to 25 ng / ml phorbol myristate-acetate (PMA) and 1 μg / ml ionomycin and various concentrations (0,50,100,200 μmol / L) Ketamine was incubated for 4 h. The percentage of Th cell subsets, the levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in supernatants and the DNA-binding activities of the intracellular transcription factors T-bet and GATA3 were then measured. RESULTS: Th1 and Th2 cell counts, IFN-γ and IL-4 levels were significantly increased by PMA and ionomycin stimulation. Ketamine dose-dependently reduced Th1 and Th2 cell counts by IFN-γ and PMA, (Th1 / Th2) and the ratio of IFN-γ and IL-4 (IFN-γ / IL-4) bet and GATA3. Ketamine reduced the T-bet and GATA3 activities and increased the ratio of T-bet to GATA3 activity (T-bet / GATA3) in a dose-dependent manner. CONCLUSION: Ketamine inhibits the differentiation of Th cells under stimulation of PMA and ionomycin, thereby inhibiting the secretion of various cytokines but increasing Th1 / Th2. The increase of Th1 / Th2 by ketamine may be related to the increase of T-bet / GATA3.