过敏性紫癜外周血单个核细胞中Tim-1、Tim-3 mRNA的表达和外周血中IL-2、IL-4含量测定
发布时间：2019-05-22 16:48
【摘要】：目的：过敏性紫癜(Henoch-Schonlein purpura，HSP)是儿童期常见的白细胞碎裂性血管炎。临床上以非血小板减少性紫癜、关节肿胀、胃肠道受累和肾炎为主要表现，多见于学龄期，冬春季为发病高峰期。HSP发病机理尚未研究透彻，多认为由多因素所致，感染、食物或者药物等都可作为致敏因素，近年来研究发现，HSP患者血清中IgA水平升高，血管壁有IgA免疫复合物（IgA-IC）沉积。Th细胞亚群功能失调在其发病机制中也起着非常重要的作用。 Th1细胞能增强巨噬细胞的吞噬作用，促进细胞免疫；Th2细胞可增强B细胞介导的体液免疫应答。Tim (T cell immunoglobin domain and mucindomain protein)家族因其特殊的结构和表达特异性，可能成为Thl/Th2特异性表面标志之一，参与T细胞的分化及免疫应答调节：Tim-1细胞主要表达在分化后的Th2细胞上，但在Th1细胞表面几乎不表达，可诱导Th2活化增殖、促进细胞因子释放、激发Th2型免疫应答；Tim-3细胞则特异性表达在分化终末期的Thl细胞上，，负调节Th1细胞介导的免疫应答。相应地，IL-2作为Th1型细胞因子、IL-4作为Th2型细胞因子，二者含量亦随之发生变化。 RT-PCR是一种基于传统PCR检测方法上的新技术，同时进行靶序列的扩增与荧光信号的检测，有效解决传统PCR定量只能在终点进行检测和传统PCR产物的污染问题，具有特异性强，操作简便，重复性好等优点。 本研究通过RT-PCR检测过敏性紫癜患者外周血单个核细胞(PBMC)中Tim-l、Tim-3mRNA的表达水平，并用ELISA方法检测外周血中IL-2、IL-4含量，探究Tim家族分子在过敏性紫癜发生中的作用，进一步阐明过敏性紫癜的发病机制。 方法：所有的实验对象均来自河北医科大学第二医院。病例组28例过敏性紫癜患者，男16例，女12例，平均年龄(19.30士9.58)岁，病程2天-3月。其中19例单纯型组（仅有皮疹），9例混合型组（有关节肿痛、腹痛或蛋白尿中的一项或多项）。对照组为20例健康正常人，经统计学分析，病例组在性别、年龄上与对照组均无统计学差异。抽取研究对象外周静脉血，采用SYBR Green I RT-PCR技术检测PBMC中Tim-1mRNA和Tim-3mRNA的表达状况,并用ELISA方法检测外周血中IL-2、IL-4含量。 数据采用SPSS13.0软件进行统计分析。正态分布数据IL-2、IL-4含量用均数±标准差(x±s)表示，组间比较采用t检验；非正态分布数据Tim-1、Tim-3mRNA表达量RQ值用中位数(四分位间距)[MD(IQR)]表示，组间比较采用Wilcoxon秩和检验，P＜0.05认为有统计学意义。 结果: 1过敏性紫癜组与健康对照组Tim-1mRNA相对表达量RQ值分别为0.974（1.108）和0.760（0.514），过敏性紫癜患者Tim-1mRNA的表达水平升高，有显著性差异（P＜0.05），而在过敏性紫癜患者中单纯型组和混合型组之间没有显著性差异（P＞0.05）。 2过敏性紫癜组与健康对照组Tim-3mRNA相对表达量RQ值分别为1.359（1.183）和1.604（1.177），两者无显著性差异（P＞0.05）。过敏性紫癜患者中单纯型组和混合型组之间亦没有显著性差异（P＞0.05）。 3过敏性紫癜组与健康对照组IL-2含量分别为188.517±12.867和202.759±20.903（pg/ml），患者组IL-2含量降低，有显著性差异（P＜0.05），过敏性紫癜组中单纯型组和混合型组IL-2含量分别为193.225±11.758和177.531±7.922（pg/ml），两者之间有显著性差异（P＜0.05）。 4过敏性紫癜组与健康对照组IL-4含量分别为73.046±8.750和62.301±11.232（pg/ml），患者组IL-4含量升高，有显著性差异（P0.05），过敏性紫癜中单纯型组和混合型组IL-4含量分别为70.400±7.642和79.218±8.591（pg/ml），两者之间有显著性差异（P＜0.05）。 结论：本研究提示HSP存在免疫功能紊乱，Th2细胞亚群占优势；Tim1mRNA表达增高，Tim蛋白可能参与HSP发病，但与疾病严重程度和疾病的发展无相关性；IL-2降低、IL-4升高在混合型组中的变化可能更明显，提示IL-2、IL-4可能与疾病严重程度密切相关。
[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卤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
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.5
本文编号：2483083
[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卤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
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.5
【参考文献】
相关期刊论文 前10条
1 师锁柱,田月,陈香美;I型纤溶酶原激活物抑制物在紫癜肾炎肾组织中的表达[J];中国组织化学与细胞化学杂志;2001年02期
2 吴舒华 ,江华;过敏性紫癜患儿血清IFN-α、IL-2、IL-4的变化[J];华中医学杂志;2002年02期
3 常红,阿爽;过敏性紫癜急性期患儿淋巴细胞凋亡特征的研究[J];山东医药;2003年14期
4 潘继豹,叶家鹤;过敏性紫癜体液免疫状态分析[J];上海医学检验杂志;2000年02期
5 董胜英,陈彤,张秋业,董增义,雷珂;过敏性紫癜病儿急性期外周血辅助性T淋巴细胞亚群功能的变化[J];齐鲁医学杂志;2004年02期
6 潘凯丽;白庆峰;黄莹;李琦;;过敏性紫癜患儿血清白细胞介素-4-、6-、8及肿瘤坏死因子-α表达的意义[J];实用儿科临床杂志;2007年21期
7 解德琼;甘华;杜晓刚;李正荣;巫江;;终末期肾病患者Th1/Th2型细胞因子的特征与外周血T细胞凋亡的关系[J];细胞与分子免疫学杂志;2006年06期
8 刘文彬,王太森,王剑峰,郑淑梅,李琴;初发过敏性紫癜患儿免疫功能研究[J];西南国防医药;2004年02期
9 李秋,杨锡强,李永柏,王莉佳;过敏性紫癜T淋巴细胞功能状态的研究[J];中华儿科杂志;2001年03期
10 高玉兴，池永学，李亚荣，尹永杰，刘禹仁;过敏性紫癜白细胞介素2水平及其受体表达[J];中华儿科杂志;1995年02期
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