摘要
目的探讨Foxp3+CD4+CD25+调节性T细胞、Foxp3mRNA在重症肌无力被动转移模型(passive transferred myasthenia gravis,PTMG)发病中的作用机制。方法将16只幼年雌性C57BL/6小鼠随机分为2组:模型组和对照组,模型组经腹腔注射含1.0mg/kgmAb35的Ringer's液0.2ml建立PTMG模型,正常对照组注射不含mAb35的Ringer's液0.2ml。采用流式细胞技术检测小鼠脾细胞中CD4+CD25+T细胞及Foxp3+CD4+CD25+Tregs含量,实时荧光定量聚合酶链反应(RT-FQ-PCR)分析2组小鼠脾细胞中Foxp3mRNA的表达,ELISA法检测2组小鼠血清白介素-2和干扰素-γ的水平。结果①模型组小鼠脾细胞中CD4+CD25+T细胞与CD4+T细胞含量的比例与对照组比较有统计学差异[(8.82±0.74)%vs(9.89±0.88)%,P<0.05];模型组Foxp3+CD4+CD25+Tregs与CD4+CD25+T细胞含量的比例与对照组比较显著降低[(6.83±1.18)%vs(8.38±0.76)%,P<0.01];脾细胞悬液中Foxp3mRNA表达量模型组低于对照组[(0.47±0.30)vs(1.53±1.19),P<0.05];模型组小鼠血清IL-2、IFN-γ水平分别为(24.41±13.83)ng/L、(142.31±6.05)ng/L,高于对照组小鼠[(12.09±2.96)ng/L、(109.36±3.64)ng/L](P<0.05)。②模型组小鼠Foxp3+CD4+CD25+Tregs数量变化与CD4+CD25+T细胞数量呈正相关(r=0.864,P<0.01),对照组小鼠Foxp3+CD4+CD25+Tregs数量变化与CD4+CD25+T细胞数量亦呈正相关(r=0.977,P<0.01)。结论Foxp3mRNA表达下调,导致Foxp3+CD4+CD25+Tregs细胞亚群表达降低,并通过上调的IL-2和IFN-γ介导了Foxp3及Tregs在重症肌无力发病机制中的作用。
Objective To study the role of Foxp3^＋ CD4^＋ CD25^＋ regulatory T cells （ Foxp3^＋ CD4^＋ CD25 ^＋Tregs） and Foxp3mRNA in pathogenesis of passive transferred myasthenia gravis （PTMG） by detecting their expression in a PTMG model. Methods Sixteen C57BL/6 young female mice were divided into model group and control group. A PTMG model was established by injecting Ringer＇s solution containing 1.0 mg/kg mAb35 into abdominal cavity. Mice in control group were injected with Ringer＇s solution not containing mAb35. Levels of CD4^＋ CD25^＋ T cells and Foxp3^＋ CD4^＋ CD25^＋ Tregs in spleen cells were measured by flow cytometry. Expression of Foxp3 mRNA in spleen cells was detected by real-time fluorescent quantitative polymerase chain reaction （RT-FQ-PCR）. Serum level of interleukin-2 and interferon-γ/was detected by ELISA. Results The ratio of CD4^＋ CD25^＋ T ceils and CD^＋ T ceils was higher in model group [ （ 8.82 ± 0. 74） % ] than in control group [ （9.89 ± 0.88 ） % ] （ P 〈 0.05 ）. The ratio of Foxp3^＋ CD4^＋ CD25^＋ Tregs and CD4 ^＋CD25^＋ T cells was significantly lower in model group [ （ 6.83 ± 1. 18 ） % ] than in control group [ （ 8.38 ± 0.76 ） % ], （ P 〈 0. 01 ）. The expression level of Foxp3 mRNA in spleen cells was significantly lower in model group （0.47 ± 0. 30） than in control group （1.53 ± 1.19） （P 〈0.05）. The serum level of IL-2 and IFN-γ/ was higher in model group （24.41 ± 13.83 and 142.31 ±6.05 ng/L） than in control group （ 12.09 ±2.96 and 109.36 ± 3.64 ng/L） （ P 〈 0. 05 ）. The change in the number of Foxp3^＋CD4^＋ CD25^＋ Tregs was positively correlated with that of CD4^＋ CD25^＋ T cells between model and control groups （ r = 0. 864 vs r = 0. 977, P 〈 0. 01 ）. Conclusion Down-regulation of Foxp3 expression can decrease the expression of Foxp3^＋ CD^＋ CD25^＋ Tregs subgroups, and up-regulation of IL-2 and IFN-γ plays an important role in the pathogenesis of Foxp3-mediated PTMG.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第4期353-356,共4页
Journal of Third Military Medical University
基金
重庆市卫生局科研基金(2008-2-169)~~
