节点文献
再生障碍性贫血患儿外周血Treg/Th17细胞失衡的研究
Study on the Imbalance of Peripheral Blood Treg/Thl7Cells in Childhood Aplastic Anemia
【作者】 王敏；
【导师】 王宁玲；
【作者基本信息】 安徽医科大学 ， 儿科学， 2012， 硕士
【摘要】 目的：通过观察再生障碍性贫血（aplastic anemia， AA）患儿外周血CD4+CD25+调节T细胞（regulatory T cells, Treg）、Th17细胞和相关细胞因子IL-17、IL-6的表达水平，探讨Treg/Th17细胞失衡在儿童再生障碍性贫血发病机制中的作用及其临床意义。方法：35例AA患儿分为重型再障组（SAA组）19例和非重型再障组（NSAA组）16例，根据疗效判定进一步分为治疗有效组13例和治疗无效组17例，同时以25例健康体检儿童作为对照组。采用流式细胞仪（flow cytometry，FCM）检测外周血Treg细胞和Th17细胞的比例，酶联免疫吸附法（ELISA）检测血浆中IL-17和IL-6的水平。统计学处理采用t检验和方差分析，相关分析采用Pearson直线相关分析。结果：1. AA患儿外周血Th17细胞[（1.45±0.28）%]、IL-17[（185.96±40.42）pg/ml]、IL-6[（20.78±5.49）pg/ml]显著高于同龄对照组[（0.45±0.10）%]、[（120.47±18.39）pg/ml]、[（10.44±2.51）pg/ml]（P<0.01），而Treg细胞比例[（4.05±1.07）%]以及Treg/Th17细胞比值（2.89±0.88）明显低于同龄对照组[（6.96±0.79）%]、（15.77±2.77）（P<0.01）。2.SAA组患儿Treg细胞比例和Treg/Th17细胞比值显著低于NSAA组，分别为[（3.37±0.79）%，(2.47±0.78)；（4.84±0.78）%,(3.39±0.75)]（P<0.01），而Th17细胞、IL-17、IL-6差异无统计学意义分别为[(1.43±0.31)%，(191.76±50.62) pg/ml,(21.65±6.26)pg/ml；（1.46±0.23）%,(179.08±23.13) pg/ml,(19.75±4.37) pg/ml]（P>0.05）。3.AA治疗有效组Th17细胞比例（0.56±0.15）%显著低于治疗无效组（1.44±0.26）%（P<0.01），而Treg细胞比例（6.63±1.01）%以及Treg/Th17细胞比值（12.92±4.72）明显高于治疗无效组（4.26±1.04）%、（3.09±1.00）（P<0.01）；治疗有效组与正常对照组之间Treg细胞、Th17细胞比例及Treg/Th17细胞比值差异无统计学意义（P>0.05）。4.AA患儿外周血Th17细胞比例与血浆IL-17和IL-6水平成正相关（r=0.67, p<0.01；r=0.57, p<0.01），而Treg细胞与IL-6呈负相关（r=-0.39, p<0.05）。结论：AA患儿Treg细胞比率的减低和Th17细胞比率的增加所致的Treg/Th17细胞失衡，可能在再障的免疫发病机制中起着重要作用。IL-17的高表达，可能是Th17细胞发挥致病作用的机制之一，而IL-6的高表达，可能是导致Treg/Th17细胞细胞失衡的原因之一。此外，Treg/Th17细胞失衡程度可能还有助于再障的病情判断及疗效评价。
【Abstract】 Objective：To investigate the levels of CD4+CD25+regulatory T cells(Treg),Th17cells and their related cytokines in the peripheral blood ofchildhood aplastic anemia(AA),and explore the roles of Treg/Th17cellimbalance in pathogenesis of AA in children and the clinical significance.Methods: There were35cases of children with AA,19patients with severeaplastic anemia(SAA),16patients with non-severe aplastic anemia(NSAA).The AA patients were further divided into effective therapy group(n=13)and ineffective treatment group(n=15).25healthy subjects were selectedas normal control group. The flow cytometry was used to evaluate theproportion of peripheral blood Treg cells and Th17cells in childhoodaplastic anemia and healthy children. The levels of plasma IL-17and IL-6were assayed by enzyme-linked immunosorbent assay(ELISA).T-test andChi-square test were used for inter-group comparison and Pearon’s linearanalysis was used for correlation analysis.Results: Compared with healthy controls, the percentages of Th17cells andthe concentration of IL-17and IL-6in the peripheral blood of childhoodaplastic anemia increased significantly [(1.45±0.28)%vs (0.45±0.10)%；（185.96±40.42）pg/ml vs（120.47±18.39）pg/ml、（20.78±5.49）pg/ml vs（10.44±2.51）pg/ml](P<0.01), while the ratio of Treg cells and Treg/Th17was markedly lower[（4.05±1.07）%vs (6.96±0.79)%；（2.89±0.88）VS（15.77±2.77）](P<0.01). The ratio of Treg cells and Treg/Th17in patients with SAA was lower than that in patients with NSAA[respectively for（3.37±0.79）%，(2.47±0.78) and （4.84±0.78）%,(3.39±0.75),P<0.01], but thelevel of Th17cells,IL-17and IL-6didn’t markedly changed [respectivelyfor (1.43±0.31)%，(191.76±50.62) pg/ml,(21.65±6.26) pg/ml and （1.46±0.23）%,(179.08±23.13) pg/ml,(19.75±4.37) pg/ml, P>0.05].Comparedwith ineffective treatment group, the proportion of Th17cells in effectivetherapy group decreased markedly,while the ratio of Treg cells andTreg/Th17increased significantly [(0.56±0.15)%vs (1.44±0.26)%；（6.63±1.01）%vs (4.26±1.04)%；（12.92±4.72）VS（3.09±1.00）](P<0.01). Therewere no significant difference in the proportion of Treg or Th17cells orthe ratio of Treg/Th17between effective therapy group and normal controlgroup (P>0.05). The proportion of Th17cells in the peripheral blood ofchildhood aplastic anemia were positively correlated to the plasma levelsof IL-17and IL-6(r=0.67,P<0.01；r=0.57,P<0.01),but the percentage of Treghad negative correlation with the plasma concentration ofIL-6(r=-0.39,P<0.05).Conclusion: The imbalance between Treg cells and Th17cells resulted fromdecreased ratio of Treg cells and increased ratio of Th17cells may playa critical role in immunological pathogenesis of childhood aplastic anemia.High expression of IL-17may be one of the mechanism of Th17cells inpathogenesis of childhood aplastic anemia,while the increase expressionof IL-6may be one of the factors causing the imbalance of Treg/Th17cells.Furthermore， the imbalance between Treg cells and Th17cells couldcontribute to predicting the disease severity and clinical evaluation ofAA.