雷帕霉素对哮喘小鼠CD4+CD25+调节性T细胞的作用
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Effect of rapamycin on CD4+CD25+ regulatory T cells in asthmatic mice
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    目的:建立哮喘小鼠动物模型并给予吸入雷帕霉素治疗,观察雷帕霉素治疗哮喘的效果及其对CD4+CD25+调节性T细胞(regulatory T cells,Treg)比例及扩增能力的影响,探讨雷帕霉素治疗哮喘的相关机制。方法:采用随机数字表法将30只雌性BALB/c 小鼠分成3组,每组10只,分别为:对照组、模型组和雷帕霉素治疗组。采用Underwood方法对小鼠肺组织进行评分;采用流式细胞术观察小鼠外周血和肺组织中CD4+CD25+Treg细胞比例;不同浓度(50、100 nmol/L)雷帕霉素处理小鼠CD4+ T细胞后,采用流式细胞术和RT-PCR方法分别检测CD4+CD25+Treg细胞形成比例和Foxp3 mRNA表达情况,观察雷帕霉素对小鼠的影响。结果:与模型组相比,雷帕霉素治疗能够显著改善小鼠肺组织病理评分情况;雷帕霉素治疗后,与模型组相比,外周血单个核细胞和肺组织中CD4+CD25+Treg均显著升高(P<0.01);50、100 mol/L雷帕霉素处理小鼠CD4+T细胞后,CD4+CD25+Treg细胞比例分别为(7.47±1.84)%和(10.93±2.74)%,与对照组[(4.84±0.76)%]相比,雷帕霉素处理可显著促进CD4+CD25+Treg细胞增殖(P<0.01);RT-PCR结果与流式细胞结果一致,雷帕霉素处理可显著促进CD4+T细胞中Foxp3的表达(P<0.01)。结论:雷帕霉素可能通过促进CD4+CD25+Treg细胞增殖,增加哮喘小鼠外周血和肺组织中CD4+CD25+Treg细胞比例,进而对小鼠哮喘发挥治疗作用。

    Abstract:

    Objective:To investigate the effect of inhaling rapamycin on CD4+CD25+ regulatory T cells (Treg) ratio and the treatment of asthma,as well as its related mechanisms,using asthmatic mice treated with inhaled rapamycin therapy. Methods:A total of 30 cases of BALB/c mice were random divided into three groups:the control group,the model group,and the rapamycin treatment group,each group had 10 cases. Underwood method was performed to estimate mouse lung tissue scores. CD4+CD25+ Treg cell ratios of peripheral blood and lung tissues were detected by flow cytometry. Different concentrations(50 and 100 nmol/L) rapamycin were used to treat mice CD4+ T cells,and CD4+CD25+ Treg cell formation ratio and Foxp3 mRNA expression were analyzed by flow cytometry and RT-PCR methods. Results:Compared with the model group,rapamycin treatment significantly improved lung tissue pathology score. CD4+CD25+ Treg percentage in the model group was (1.94 ± 0.37)% and (2.74±0.42)% in peripheral blood mononuclear cells(PBMCs) and lung tissues,respectively,after rapamycin treatment,the CD4+CD25+ Treg percentage was(5.11±0.64)% and(2.74±0.42)% in PBMCs and lung tissues,respectively. After treated with different concentrations of rapamycin,CD4+CD25+ Treg ratio in the control group was(4.84 ± 0.76)%,and in the 50 nmol/L and the 100 nmol/L rapamycin groups was(7.47±1.84)% and(10.93±2.74)%,respectively. RT-PCR results were consistent with the results of flow cytometry staining,rapamycin treatment significantly promoted CD4 T cells Foxp3 expression (P<0.01). Conclusion:By promoting CD4+CD25+ Treg cell proliferation,rapamycin can increase CD4+CD25+ Treg cell ratio of peripheral blood and lung tissues in asthmatic mice,and thus exert a therapeutic effect on asthma.

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黄文军,张世应,胡小平.雷帕霉素对哮喘小鼠CD4+CD25+调节性T细胞的作用[J].南京医科大学学报(自然科学版),2017,(3):307-311

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  • 收稿日期:2015-11-09
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  • 在线发布日期: 2017-04-07
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