银耳多糖改善脓毒症小鼠调节性T细胞的免疫活性

发布时间：2018-03-02 12:39

  本文选题：银耳多糖　切入点：调节性T淋巴细胞　出处：《中国免疫学杂志》2016年03期 　论文类型：期刊论文

【摘要】：目的:探讨银耳多糖(TPS)对脓毒症小鼠外周血调节性T细胞免疫调节的作用。方法:雄性BALB/c小鼠(20±1)g,采用烧伤后腹腔注射铜绿假单胞菌制作脓毒症并予以TPS干预,随机分成5组,分别为正常对照组(未做任何处理),未刺激组(烧伤后铜绿假单胞菌感染未经TPS处理组),TPS低剂量组(烧伤后铜绿假单胞菌感染+50 mg/kg治疗组),TPS中剂量组(烧伤后铜绿假单胞菌感染+100 mg/kg治疗组),TPS高剂量组(烧伤后铜绿假单胞菌感染+200 mg/kg治疗组),从各组的外周血中分离调节性T细胞,并体外培养。自烧伤即刻(PBD0)起,至烧伤后4 d每天监测培养上清液中IL-10、IFN-γ、IL-4的水平。磁珠孵育和磁性分离小鼠外周血的调节性T细胞( CD4~+CD25~(high)Tregs)和CD4~+T细胞,分别加入TPS后进行培养,以不加TPS的细胞作为对照,流式细胞仪分析 CD4~+CD25~(high)Tregs的表型,ELISA法检测细胞因子的表达。结果:与对照组相比,TPS能显著降低未刺激组小鼠的IL-10和IL-4的分泌,并显著增加IFN-γ的分泌,并且IL-10的分泌水平与TPS浓度呈剂量效应关系。体外培养试验中,与正常对照组相比,未刺激组的CD4~+T细胞增殖和IFN-γ水平显著降低(P0.05),IL-4水平显著升高(P0.05);与未刺激组相比,TPS刺激组的CD4~+T细胞增殖和IFN-γ水平显著升高(P0.05),IL-4水平显著降低(P0.05);与未刺激组相比,TPS刺激+抗体1组CD4~+T细胞增殖和IFN-γ水平显著升高(P0.05),IL-4水平显著降低(P0.05);与未刺激组相比,TPS刺激+抗体2组的CD4~+T细胞增殖和IFN-γ、IL-4水平无显著性差异。结论:TPS可以通过降低IL-10的分泌抑制 CD4~+CD25~(high)Tregs对CD4~+T淋巴细胞增殖和极化的影响,并诱导CD4~+T淋巴细胞向Th1分型,从而使机体免疫活性增强。
[Abstract]:Objective: to investigate the effect of Tremella polysaccharide (TP) on the immune regulation of regulatory T cells in peripheral blood of septic mice. Methods: male BALB/c mice were injected intraperitoneally with Pseudomonas aeruginosa to make sepsis and treated with TPS after burn, and were randomly divided into 5 groups. They were normal control group (without any treatment) and unstimulated group (postburn Pseudomonas aeruginosa infection without TPS treatment group with low dose (50 mg/kg post-burn Pseudomonas aeruginosa infection treatment group). Pseudomonas aeruginosa infection for 100 mg/kg was used to isolate regulatory T cells from peripheral blood of patients with high dose of Pseudomonas aeruginosa infection (200 mg/kg after burn). The levels of IL-10 IFN- 纬 IL-4 in the supernatant were monitored daily from immediate burn to 4 days after burn. The regulatory T cells (CD _ 4 ~ CD _ (25) T _ (+)) and CD _ 4 ~ T cells were incubated with magnetic beads and separated from the peripheral blood of mice. The expression of cytokines was detected by flow cytometry in cells without TPS. Results: compared with the control group, TPS could significantly decrease the secretion of IL-10 and IL-4 and increase the secretion of IFN- 纬 in unstimulated mice. And the level of IL-10 secretion was dose-dependent with the concentration of TPS. In vitro culture test, compared with the normal control group, The proliferation of CD4 ~ T cells and the level of IFN- 纬 in unstimulated group were significantly lower than those in unstimulated group. The level of IL-4 in P0.05 was significantly higher than that in unstimulated group, while the proliferation of CD4 ~ T cell and the level of IFN- 纬 in TPS stimulated group were significantly higher than that in unstimulated group. The proliferation of CD4 ~ T cells and the level of IFN- 纬 increased significantly in group 1, and the level of IL-4 decreased significantly in group P0.05, but there was no significant difference in proliferation of CD4T cells and IL-4 level of IFN- 纬 IL-4 in TPS stimulated antibody group 2 compared with that in group 2. Conclusion the level of TPS can be reduced by decreasing the level of IL-4. The effect of IL-10 secretion on the proliferation and polarization of CD4 ~ T lymphocytes was inhibited by CD4 ~ CD25~(high)Tregs. CD4 ~ T lymphocytes were induced to type to Th1, and the immune activity was enhanced.
【作者单位】： 煤炭总医院肾内科;解放军总医院第一附属医院全军烧伤研究所;
【基金】：国家自然科学基金(81000847,81272140) 中国博士后面上项目(201150M1530) 中国博士后第五批特别资助项目(2012T50863)
【分类号】：R459.7
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