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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 m RNA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。
OBJECTIVE: To establish an animal model of asthma in mice and receive inhaled rapamycin to observe the effect of rapamycin in treating asthma and its effect on the ratio of CD4 ~ + CD25 ~ + regulatory T cells (Tregs) and their amplification Ability to explore the mechanism of rapamycin in the treatment of asthma. Methods: Thirty female BALB / c mice were randomly divided into three groups (n = 10): control group, model group and rapamycin group. The lung tissues of mice were scored under the Underwood method. The proportion of CD4 ~ + CD25 ~ + Treg cells in peripheral blood and lung tissues of mice was observed by flow cytometry. Different concentrations of rapamycin (50,100 nmol / L) The percentage of CD4 ~ + CD25 ~ + Treg cells and the expression of Foxp3 mRNA were detected by flow cytometry and RT-PCR respectively. The effects of rapamycin on mice were observed. Results: Compared with the model group, rapamycin treatment could significantly improve the pathological score of lung tissue in mice. After rapamycin treatment, compared with the model group, the levels of CD4 ~ + CD25 in peripheral blood mononuclear cells and lung tissue ~ + Treg were significantly increased (P <0.01). The proportion of CD4 ~ + CD25 ~ + Treg cells in CD4 + T cells treated with 50,100 mol / L rapamycin was (7.47 ± 1.84)% and (10.93 ± 2.74)%, rapamycin treatment could significantly promote the proliferation of CD4 ~ + CD25 ~ + Treg cells (P <0.01) compared with the control group [(4.84 ± 0.76)%] The expression of Foxp3 in CD4 + T cells was significantly enhanced by rapamycin treatment (P <0.01). Conclusion: Rapamycin can increase the percentage of CD4 ~ + CD25 ~ + Treg cells in peripheral blood and lung tissue of asthmatic mice by promoting the proliferation of CD4 ~ + CD25 ~ + Treg cells and then treating mice with asthma.