本文選題：再生障礙性貧血　切入點：調(diào)節(jié)性T細胞　出處：《鄭州大學(xué)》2012年碩士論文

【摘要】：再生障礙性貧血(簡稱再障,Aplastic Anemia,AA)是由理化、生物、藥物和其他不明因素引起的骨髓造血干細胞數(shù)量下降、造血微環(huán)境損傷導(dǎo)致骨髓造血功能衰竭,出現(xiàn)以全血細胞(白細胞、紅細胞、血小板)減少為主要臨床表現(xiàn)的疾病。該疾病對患者的危害大,病死率高,治療療程長,療效差,在兒童中尤甚。因此,白再障被發(fā)現(xiàn)并報道以來,人們對其發(fā)病機制的探討從未間斷過,但至今依然不是很明了。不過,大多數(shù)國內(nèi)外學(xué)者們研究認為免疫異常在再障的發(fā)病中起著重要的作用。 近年來的研究熱點集中在T調(diào)節(jié)細胞(regulatory T cell,Treg cell)及Th17細胞的研究上。T調(diào)節(jié)細胞是一群具有免疫抑制功能的細胞,增殖能力低,可以通過識別自身抗原來抑制具有潛在傷害性的自身反應(yīng)性T細胞。該細胞可以控制免疫應(yīng)答強度,維持自身免疫穩(wěn)定,在自身免疫性疾病的發(fā)病中起著重要的作用。叉頭／翼狀螺旋轉(zhuǎn)錄因子(foxhead/winged-helix transprcription factor p3,Foxp3)是CD4+CD25+Treg中得以證實的的特異性標記物,對CD4+CD25+Treg的分化發(fā)育和發(fā)揮功能起著重要的作用,其在自身免疫性疾病的發(fā)生、發(fā)展中亦發(fā)揮著重要的作用。 另外,一種新型的T細胞(Th17細胞)所分泌的細胞因子IL-17也被證實在自身免疫性疾病中扮演著重要角色。IL-17在發(fā)揮作用時往往是與受體結(jié)合,而后被激活,可以使一系列的趨化因子、粘附分子和集落刺激因子進行表達或釋放,從而影響自身免疫、腫瘤和感染的病理過程。此外,IL-17還能夠使人類造血祖細胞的增殖過程受到抑制。 環(huán)孢素(Cyclosporine A,CsA)作為一種免疫抑制劑,它能有效的抑制T細胞介導(dǎo)的細胞免疫反應(yīng),阻斷T細胞的增殖和分化。 綜上分析,T調(diào)節(jié)細胞及其細胞因子Foxp3、Th17細胞及IL-17A基因均在再生障礙性貧血的發(fā)生、發(fā)展中發(fā)揮著重要的作用。而環(huán)孢素作為免疫抑制劑,或許會對T調(diào)節(jié)細胞及其細胞因子Foxp3、Th17細胞及IL-17A基因的表達有一定的影響�；诖�,我們設(shè)計如下實驗。 目的 觀察環(huán)孢素對慢性再障患兒外周血T調(diào)節(jié)細胞、Foxp3及IL-17基因的影響,為再障發(fā)病機制的研究及治療靶點的尋求提供新的理論依據(jù)。 材料與方法 收集鄭州大學(xué)第三附屬醫(yī)院及鄭州大學(xué)第一附屬醫(yī)院確診為慢性再障的50例患兒,這些患兒均未進行任何治療,將其隨機分為環(huán)孢素治療組及非環(huán)孢素治療組,分別給予治療6個月。同時收集同期鄭州大學(xué)第三附屬醫(yī)院健康體檢兒童作為正常對照組。即研究對象可分三組：CsA治療組(30例)、非CsA治療組(20例)和正常對照組(15例)。三組之間性別、年齡差異無統(tǒng)計學(xué)意義。應(yīng)用流式細胞術(shù)分別檢測各組患兒T調(diào)節(jié)細胞(CD4+CD25+CD127lowTreg)在外周血的表達,并采用實時熒光定量PCR檢測Foxp3mRNA及IL-17A mRNA在各組患兒外周血中的表達。采用SPSS17.0統(tǒng)計軟件分析,數(shù)據(jù)用均數(shù)±標準差((?)±s)表示,多組樣本均數(shù)比較采用方差分析檢驗,組間兩兩比較采用LSD檢驗,以P0.05表示差異有統(tǒng)計學(xué)意義。 結(jié)果 1.CD4+CD25+CD127low Treg表達水平：再障CsA治療組為(5.31±1.18),明顯高于非CsA治療組(4.56±0.95),接近于正常對照組(5.91±1.51)； 2.Foxp3mRNA水平：再障CsA治療組為(1.01±0.26),與對照組(1.17±0.32)相比,差異沒有統(tǒng)計學(xué)意義,而非CsA治療組(0.79±0.34)則明顯低于CsA治療組及正常對照組； 3.IL-17A mRNA水平：再障CsA治療組水平接近對照組,兩組比較無明顯差異(P0.05)；非CsA治療組的表達水平均顯著高于對照組(P0.05)；CsA治療組的表達水平均低于非CsA治療組,差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論 1.環(huán)孢素可以促進再障患兒外周血CD4+CD25+CD127lowTreg及Foxp3mRNA表達水平上升；促進IL-17AmRNA水平下降。 2.T調(diào)節(jié)細胞、Foxp3及IL-17參與了再障的發(fā)生、發(fā)展過程。
[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.

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.5

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