环孢素对慢性再生障碍性贫血患儿外周血T调节细胞及IL-17基因表达的影响

发布时间：2018-03-29 07:32

本文选题：再生障碍性贫血　切入点：调节性T细胞　出处：《郑州大学》2012年硕士论文

【摘要】：再生障碍性贫血(简称再障,Aplastic Anemia,AA)是由理化、生物、药物和其他不明因素引起的骨髓造血干细胞数量下降、造血微环境损伤导致骨髓造血功能衰竭,出现以全血细胞(白细胞、红细胞、血小板)减少为主要临床表现的疾病。该疾病对患者的危害大,病死率高,治疗疗程长,疗效差,在儿童中尤甚。因此,白再障被发现并报道以来,人们对其发病机制的探讨从未间断过,但至今依然不是很明了。不过,大多数国内外学者们研究认为免疫异常在再障的发病中起着重要的作用。近年来的研究热点集中在T调节细胞(regulatory T cell,Treg cell)及Th17细胞的研究上。T调节细胞是一群具有免疫抑制功能的细胞,增殖能力低,可以通过识别自身抗原来抑制具有潜在伤害性的自身反应性T细胞。该细胞可以控制免疫应答强度,维持自身免疫稳定,在自身免疫性疾病的发病中起着重要的作用。叉头／翼状螺旋转录因子(foxhead/winged-helix transprcription factor p3,Foxp3)是CD4+CD25+Treg中得以证实的的特异性标记物,对CD4+CD25+Treg的分化发育和发挥功能起着重要的作用,其在自身免疫性疾病的发生、发展中亦发挥着重要的作用。另外,一种新型的T细胞(Th17细胞)所分泌的细胞因子IL-17也被证实在自身免疫性疾病中扮演着重要角色。IL-17在发挥作用时往往是与受体结合,而后被激活,可以使一系列的趋化因子、粘附分子和集落刺激因子进行表达或释放,从而影响自身免疫、肿瘤和感染的病理过程。此外,IL-17还能够使人类造血祖细胞的增殖过程受到抑制。环孢素(Cyclosporine A,CsA)作为一种免疫抑制剂,它能有效的抑制T细胞介导的细胞免疫反应,阻断T细胞的增殖和分化。综上分析,T调节细胞及其细胞因子Foxp3、Th17细胞及IL-17A基因均在再生障碍性贫血的发生、发展中发挥着重要的作用。而环孢素作为免疫抑制剂,或许会对T调节细胞及其细胞因子Foxp3、Th17细胞及IL-17A基因的表达有一定的影响。基于此,我们设计如下实验。目的观察环孢素对慢性再障患儿外周血T调节细胞、Foxp3及IL-17基因的影响,为再障发病机制的研究及治疗靶点的寻求提供新的理论依据。材料与方法收集郑州大学第三附属医院及郑州大学第一附属医院确诊为慢性再障的50例患儿,这些患儿均未进行任何治疗,将其随机分为环孢素治疗组及非环孢素治疗组,分别给予治疗6个月。同时收集同期郑州大学第三附属医院健康体检儿童作为正常对照组。即研究对象可分三组：CsA治疗组(30例)、非CsA治疗组(20例)和正常对照组(15例)。三组之间性别、年龄差异无统计学意义。应用流式细胞术分别检测各组患儿T调节细胞(CD4+CD25+CD127lowTreg)在外周血的表达,并采用实时荧光定量PCR检测Foxp3mRNA及IL-17A mRNA在各组患儿外周血中的表达。采用SPSS17.0统计软件分析,数据用均数±标准差((?)±s)表示,多组样本均数比较采用方差分析检验,组间两两比较采用LSD检验,以P0.05表示差异有统计学意义。结果 1.CD4+CD25+CD127low Treg表达水平：再障CsA治疗组为(5.31±1.18),明显高于非CsA治疗组(4.56±0.95),接近于正常对照组(5.91±1.51)； 2.Foxp3mRNA水平：再障CsA治疗组为(1.01±0.26),与对照组(1.17±0.32)相比,差异没有统计学意义,而非CsA治疗组(0.79±0.34)则明显低于CsA治疗组及正常对照组； 3.IL-17A mRNA水平：再障CsA治疗组水平接近对照组,两组比较无明显差异(P0.05)；非CsA治疗组的表达水平均显著高于对照组(P0.05)；CsA治疗组的表达水平均低于非CsA治疗组,差异有统计学意义(P0.05)。结论 1.环孢素可以促进再障患儿外周血CD4+CD25+CD127lowTreg及Foxp3mRNA表达水平上升；促进IL-17AmRNA水平下降。 2.T调节细胞、Foxp3及IL-17参与了再障的发生、发展过程。
[Abstract]:Aplastic anemia (AA, Aplastic, Anemia, AA) is a chemical, biological, pharmaceutical and other causes of bone marrow hematopoietic stem cells of unknown factors lead to a decline in the number of bone marrow failure damage of hematopoietic microenvironment, in whole blood cells (white blood cells, red blood cells, platelets) decreased as the main clinical manifestations the disease. The disease harm to patients, high fatality rate, long course of treatment, curative effect is poor, especially in children. Therefore, since aplastic anemia was found and reported, the discussion about its pathogenesis has never been interrupted, but until today is still not very clear. However, most scholars think immune abnormalities in the pathogenesis of aplastic anemia in plays an important role.
The hot research in recent years focused on regulatory T cells (regulatory T cell, Treg cell) and Th17 cell research on.T regulatory cells are a group of immune suppression of cell proliferation ability is low, can be identified by its original anti inhibit potentially harmful self reactive T cells. To control the strength of the immune response cells can maintain immune stability, plays an important role in the pathogenesis of autoimmune diseases. The forkhead / winged helix transcription factor (foxhead/winged-helix transprcription, factor P3, Foxp3) is to confirm the specific marker of CD4+CD25+Treg, CD4+CD25+Treg on differentiation and function plays an important role in the in autoimmune diseases, the development also play an important role.
In addition, a new type of T cells (Th17 cells) cell factor IL-17 secretion has been shown to play in autoimmune disease has an important role to play a role in.IL-17 is often associated with receptor binding, then activated, can make a series of chemokines, adhesion molecules and colony-stimulating factor expression or release, thus affecting the pathological process of autoimmunity, cancer and infection. In addition, IL-17 can also make the proliferation of human hematopoietic progenitor cells was inhibited.
Cyclosporine A (CsA), as an immunosuppressant, can effectively inhibit the cellular immune response mediated by T cells and block the proliferation and differentiation of T cells.
In conclusion, regulation of T cells and Foxp3 cells, Th17 and IL-17A genes were in aplastic anemia, plays an important role in the development. And cyclosporine as immunosuppressive agents, may be the regulation of T cells and Foxp3, have a certain effect on the expression of Th17 cells and IL-17A gene based on this., we designed the following experiment.
objective
The effects of cyclosporine on chronic aplastic anemia in children with peripheral blood T regulatory cells, the effects of Foxp3 and IL-17 gene, and provide a new theoretical basis for research and therapeutic target in the pathogenesis of aplastic anemia.
Materials and methods
50 cases were collected in First Affiliated Hospital of the Third Affiliated Hospital of Zhengzhou University and the Zhengzhou University for chronic aplastic anemia, these patients were not treated, they were randomly divided into treatment group and non cyclosporine CsA treatment group, were treated for 6 months. At the same time to collect the Affiliated Hospital of Zhengzhou University in the same period of third healthy children as normal control group. The object of study can be divided into three groups: CsA treatment group (30 cases) and non CsA group (20 cases) and control group (15 cases). The sex between the three groups, no significant difference in age. The children were detected T regulatory cells by flow cytometry (CD4+CD25+CD127lowTreg) expression in peripheral blood and, using real-time PCR to detect the expressions of Foxp3mRNA and IL-17A mRNA in serum in peripheral blood. The data were analyzed by statistical software SPSS17.0, with the mean and standard deviation ((?) + s) said that many groups like The average numbers were compared by analysis of variance analysis, and 22 of the groups were compared by LSD test, and the difference was statistically significant with P0.05.
Result
1.CD4+CD25+CD127low Treg: the expression level of CsA in aplastic anemia treatment group (5.31 + 1.18), was significantly higher than that in non CsA group (4.56 + 0.95), close to the normal control group (5.91 + 1.51);
2.Foxp3mRNA: AA CsA treated group (1.01 + 0.26), and control group (1.17 + 0.32) compared to the difference was not statistically significant, and the non CsA group (0.79 + 0.34) was significantly lower than that of CsA group and normal control group;
3.IL-17A mRNA: AA CsA treatment group close to the level of control group, the two groups had no significant difference (P0.05); the expression level of CsA in non treatment group were significantly higher than the control group (P0.05); the expression level of CsA in the treatment group were lower than CsA treatment group, the difference was statistically significant (P0.05).
conclusion
1. cyclosporine can promote the peripheral blood of children with aplastic anemia and CD4+CD25+CD127lowTreg Foxp3mRNA expression level increased; promote the level of IL-17AmRNA decreased.
2.T Foxp3 and IL-17 regulatory cells in aplastic anemia, the development process.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.5

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