左归丸对小鼠CD4^+CD25^+调节性T细胞的影响

Effects of Zuogui pills on CD4^+ CD25^+ regulatory T cells of mice

目的:观察左归丸制剂对小鼠脾细胞中Treg亚群及其相关细胞因子表达的影响。方法:BALB/c小鼠给予不同剂量左归丸处理后,利用流式细胞术检测小鼠脾细胞中Treg亚群(CD4+/CD25+)的变化;采用RT-PCR法检测小鼠脾细胞中IL-10、TGF-β、IFN-γ及Foxp3的表达水平;采用ELISA法检测小鼠外周血中IFN-γ的分泌水平。结果:小剂量左归丸对小鼠脾脏Treg亚群及相关细胞因子的表达没有显著影响;中剂量左归丸可明显上调小鼠脾脏Treg亚群比例(P<0.05),提高Treg特异性胞内信号Foxp3及相关细胞因子IL-10、TGF-β的转录水平,同时抑制IFN-γ的表达;大剂量左归丸可显著降低Treg细胞亚群比例,对Foxp3I、L-10、TGF-β、IFN-γ的表达均有明显的抑制作用(P<0.05)。随着中、大剂量左归丸处理小鼠IFN-γ的转录下调,血清中IFN-γ的水平也明显下降(P<0.05)。结论:左归丸可上调Treg细胞及相关细胞因子的表达水平,抑制IFN-γ的表达,但这种免疫效应有剂量限制性,大剂量应用时显示抑制作用,提示左归丸对Treg亚群有剂量依赖性的双向调节作用。

Objective： To investigate the effects of Zuogui pills on CD4^＋ CD25^＋ regulatory T subpopulation （Treg） and on the expression of related cytokines of spleen in mice. Methods： Inbred BALB/c mice treated with different doses of Zuogui pills, flowcytomytre, RT-PCR and ELISA were used to analyze Treg subpopulation, the levels of IL-10, TGF-β, IFN-γ, Foxp3 transcriptions and the production of IFN-γ in peripheral blood. Results： After treatment with low dose of Zuogui pills, Treg subpopulation and the levels of IL-10, TGF-β, IFN-γ, Foxp3 transcriptions were not different from normal mice. Mid-dose of Zuogui pills treatment obviously up-regulated the proportion of Treg （P〈0.05）, increased the expressions of Foxp3, IL-10, TGF-β and decreased the production of IFN-γ. High-dose of Zuogui pills treatment decreased the proportionality of Treg and inhibited the expression of Foxp3, IL-10, TGF-β, IFN-γ （P 〈 0.05） significantly. Following inhibition of IFN-γ transcription, the production of IFN-γ in serum （pg/ml） was also depressed after treat-merits with mid-doses and high-dosea of Zuogui pills. Conclusion： Zuogui pills can increase the proportion of CD4^＋ CD25^＋ Treg cells, and the application in high-dose of Zuogui pills showed opposite regulation compared with mld-dose. This study showed the two-way immunological regulation of Zuogui pills on Treg population is dose-dependent.