【摘要】：目的：過敏性紫癜(Henoch-Schonlein purpura，HSP)是兒童期常見的白細(xì)胞碎裂性血管炎。臨床上以非血小板減少性紫癜、關(guān)節(jié)腫脹、胃腸道受累和腎炎為主要表現(xiàn)，多見于學(xué)齡期，冬春季為發(fā)病高峰期。HSP發(fā)病機(jī)理尚未研究透徹，多認(rèn)為由多因素所致，感染、食物或者藥物等都可作為致敏因素，近年來研究發(fā)現(xiàn)，HSP患者血清中IgA水平升高，血管壁有IgA免疫復(fù)合物（IgA-IC）沉積。Th細(xì)胞亞群功能失調(diào)在其發(fā)病機(jī)制中也起著非常重要的作用。 Th1細(xì)胞能增強(qiáng)巨噬細(xì)胞的吞噬作用，促進(jìn)細(xì)胞免疫；Th2細(xì)胞可增強(qiáng)B細(xì)胞介導(dǎo)的體液免疫應(yīng)答。Tim (T cell immunoglobin domain and mucindomain protein)家族因其特殊的結(jié)構(gòu)和表達(dá)特異性，可能成為Thl/Th2特異性表面標(biāo)志之一，參與T細(xì)胞的分化及免疫應(yīng)答調(diào)節(jié)：Tim-1細(xì)胞主要表達(dá)在分化后的Th2細(xì)胞上，但在Th1細(xì)胞表面幾乎不表達(dá)，可誘導(dǎo)Th2活化增殖、促進(jìn)細(xì)胞因子釋放、激發(fā)Th2型免疫應(yīng)答；Tim-3細(xì)胞則特異性表達(dá)在分化終末期的Thl細(xì)胞上，，負(fù)調(diào)節(jié)Th1細(xì)胞介導(dǎo)的免疫應(yīng)答。相應(yīng)地，IL-2作為Th1型細(xì)胞因子、IL-4作為Th2型細(xì)胞因子，二者含量亦隨之發(fā)生變化。 RT-PCR是一種基于傳統(tǒng)PCR檢測(cè)方法上的新技術(shù)，同時(shí)進(jìn)行靶序列的擴(kuò)增與熒光信號(hào)的檢測(cè)，有效解決傳統(tǒng)PCR定量只能在終點(diǎn)進(jìn)行檢測(cè)和傳統(tǒng)PCR產(chǎn)物的污染問題，具有特異性強(qiáng)，操作簡(jiǎn)便，重復(fù)性好等優(yōu)點(diǎn)。 本研究通過RT-PCR檢測(cè)過敏性紫癜患者外周血單個(gè)核細(xì)胞(PBMC)中Tim-l、Tim-3mRNA的表達(dá)水平，并用ELISA方法檢測(cè)外周血中IL-2、IL-4含量，探究Tim家族分子在過敏性紫癜發(fā)生中的作用，進(jìn)一步闡明過敏性紫癜的發(fā)病機(jī)制。 方法：所有的實(shí)驗(yàn)對(duì)象均來自河北醫(yī)科大學(xué)第二醫(yī)院。病例組28例過敏性紫癜患者，男16例，女12例，平均年齡(19.30士9.58)歲，病程2天-3月。其中19例單純型組（僅有皮疹），9例混合型組（有關(guān)節(jié)腫痛、腹痛或蛋白尿中的一項(xiàng)或多項(xiàng)）。對(duì)照組為20例健康正常人，經(jīng)統(tǒng)計(jì)學(xué)分析，病例組在性別、年齡上與對(duì)照組均無統(tǒng)計(jì)學(xué)差異。抽取研究對(duì)象外周靜脈血，采用SYBR Green I RT-PCR技術(shù)檢測(cè)PBMC中Tim-1mRNA和Tim-3mRNA的表達(dá)狀況,并用ELISA方法檢測(cè)外周血中IL-2、IL-4含量。 數(shù)據(jù)采用SPSS13.0軟件進(jìn)行統(tǒng)計(jì)分析。正態(tài)分布數(shù)據(jù)IL-2、IL-4含量用均數(shù)±標(biāo)準(zhǔn)差(x±s)表示，組間比較采用t檢驗(yàn)；非正態(tài)分布數(shù)據(jù)Tim-1、Tim-3mRNA表達(dá)量RQ值用中位數(shù)(四分位間距)[MD(IQR)]表示，組間比較采用Wilcoxon秩和檢驗(yàn)，P＜0.05認(rèn)為有統(tǒng)計(jì)學(xué)意義。 結(jié)果: 1過敏性紫癜組與健康對(duì)照組Tim-1mRNA相對(duì)表達(dá)量RQ值分別為0.974（1.108）和0.760（0.514），過敏性紫癜患者Tim-1mRNA的表達(dá)水平升高，有顯著性差異（P＜0.05），而在過敏性紫癜患者中單純型組和混合型組之間沒有顯著性差異（P＞0.05）。 2過敏性紫癜組與健康對(duì)照組Tim-3mRNA相對(duì)表達(dá)量RQ值分別為1.359（1.183）和1.604（1.177），兩者無顯著性差異（P＞0.05）。過敏性紫癜患者中單純型組和混合型組之間亦沒有顯著性差異（P＞0.05）。 3過敏性紫癜組與健康對(duì)照組IL-2含量分別為188.517±12.867和202.759±20.903（pg/ml），患者組IL-2含量降低，有顯著性差異（P＜0.05），過敏性紫癜組中單純型組和混合型組IL-2含量分別為193.225±11.758和177.531±7.922（pg/ml），兩者之間有顯著性差異（P＜0.05）。 4過敏性紫癜組與健康對(duì)照組IL-4含量分別為73.046±8.750和62.301±11.232（pg/ml），患者組IL-4含量升高，有顯著性差異（P0.05），過敏性紫癜中單純型組和混合型組IL-4含量分別為70.400±7.642和79.218±8.591（pg/ml），兩者之間有顯著性差異（P＜0.05）。 結(jié)論：本研究提示HSP存在免疫功能紊亂，Th2細(xì)胞亞群占優(yōu)勢(shì)；Tim1mRNA表達(dá)增高，Tim蛋白可能參與HSP發(fā)病，但與疾病嚴(yán)重程度和疾病的發(fā)展無相關(guān)性；IL-2降低、IL-4升高在混合型組中的變化可能更明顯，提示IL-2、IL-4可能與疾病嚴(yán)重程度密切相關(guān)。
[Abstract]:Objective: Henoch-Schonlein purpuura (HSP) is a common leucocytic vasculitis in childhood. It is mainly characterized by non-thrombocytopenic purpura, joint swelling, gastrointestinal involvement and nephritis. The pathogenesis of HSP has not been studied thoroughly and many factors, such as infection, food or drug, can be used as a sensitizing factor. In recent years, the level of IgA in the serum of HSP patients is increased, and the blood vessel wall has IgA-IC (IgA-IC) deposition. The dysfunction of Th cell subpopulation plays a very important role in its pathogenesis. Th1 cells can enhance the phagocytosis of macrophages, promote cellular immunity, and Th2 cells can enhance the humoral immunity mediated by B cells. A. Tim (T cell immoglobulin domain and mucin protein) family may be one of the specific surface markers of Thl/ Th2 for its special structure and expression, and it can be involved in the differentiation of T cells and the regulation of immune response: Tim-1 cells are mainly expressed on the differentiated Th2 cells, but the surface of the Th1 cells is almost impossible. up to, the Th2-activated proliferation can be induced, the cytokine release is promoted, the Th2-type immune response is excited, and the Tim-3 cell is specifically expressed on the Thl cell with the end stage of differentiation, and the negative regulation of the Th1 cell-mediated immunity A. As a result, IL-2, as a Th1-type cytokine and IL-4 as a Th2-type cytokine, also changes in the content of IL-2. The RT-PCR is a new technology based on the traditional PCR detection method, and the detection of the target sequence and the detection of the fluorescent signal can be carried out at the same time, so that the detection of the traditional PCR can only be carried out at the end point and the pollution problem of the traditional PCR product is solved, the method has the advantages of strong specificity, simple and convenient operation and good repeatability. The expression level of Tim-l, Tim-3 mRNA in peripheral blood mononuclear cells (PBMC) of patients with Henoch-Schonlein purpura was detected by RT-PCR and the content of IL-2 and IL-4 in peripheral blood was detected by ELISA. The mechanism of the pathogenesis. Methods: All the experimental subjects were from Hebei Medical University. There were 28 cases of allergic purpura,16 males and 12 females, with an average age of 19.30 (9.58). The course of the course was 2 days to 3 months. Of these,19 simple groups (only a rash) and 9 mixed groups (with joint swelling and pain, abdominal pain or proteinuria) One or more of the 20 healthy controls in the control group were statistically analyzed and the case group was in gender, age, and control group The expression of Tim-1 mRNA and Tim-3 mRNA in PBMC was detected by SYBR Green I RT-PCR, and IL-2 in peripheral blood was detected by ELISA. And IL-4 content. The data is SPSS13. The statistical analysis was performed by 0 software. The normal distribution data IL-2, IL-4 content was expressed by mean square standard deviation (x% s), and t-test was used between the groups; the non-normal distribution data, Tim-1, and Tim-3 mRNA expression, were expressed as median (quartile spacing)[MD (IQR)], and Wil was used between the groups. coxon rank sum test, P <0.0 5. Results: The relative expression of Tim-1 mRNA was 0.974 (1.108) and 0.760 (0.514), and the expression level of Tim-1 mRNA in Henoch-Schonlein purpura group and healthy control group were 0.760 (0.514) and 0.760 (0.514), respectively. There was no significant difference (P <0.05), and there was no difference between the simple group and the mixed group in the patients with Henoch-Schonlein purpura. The relative expression of Tim-3 mRNA was 1.359 (1.183) and 1.604 (1.177), respectively. There was no significant difference between the simple group and the mixed group in the patients with Henoch-Schonlein purpura (P> 0.05). There was no significant difference (P> 0.05). The levels of IL-2 in the allergic purpura group and the healthy control group were 188.517-12.867 and 202.759-20.903 (pg/ ml), respectively. .531-7.922 (pg/ ml), two The levels of IL-4 in the Henoch-Schonlein purpura group and the healthy control group were 73.046-8.750 and 62.301-11.232 (pg/ ml), respectively. 79.218鹵8.591(pg/ml) Conclusion: In this study, there was a significant difference between the two groups (P <0.05). Conclusion: The present study suggests that there is an immune function disorder in HSP, the subpopulation of Th2 cells is dominant, the expression of Tim1mRNA is increased, and Tim protein may be involved in the pathogenesis of HSP, but not related to the severity of the disease and the development of the disease, and the decrease of IL-2 and the increase of IL-4. the change in the hybrid group may be more pronounced, prompting i
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R725.5

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