IgA肾病组织蛋白质组和尿液蛋白质组研究
发布时间：2018-11-28 08:33
【摘要】：背景和目的:IgA肾病(IgA nephropathy, IgAN)是目前世界范围内最常见的原发性肾小球疾病,是引起终末期肾衰竭的重要原因之一,30%～40%的IgAN患者在诊断该病后20年内发展成终末期肾病(end stage renal disease,ESRD),给家庭和社会带来了沉重的负担。IgAN的确诊目前仍依赖于肾活检,但肾穿刺活检为有创性检查,存在一定风险和禁忌证。IgAN起病隐匿,易导致诊治不及时,因此早期筛查、诊治对于防控,减少ESRD患者数量尤为重要。因此,探寻IgAN的发病机制和早期无创生物标志物具有重要意义。本研究采用Raybiotech细胞因子抗体芯片技术对IgAN患者和对照组肾脏组织进行检测;采用高分辨质谱技术对IgAN患者、原发性膜性肾病(idiopathic membranous nephropathy,IMN)患者和正常人群尿液进行对照检测分析;筛选IgAN的生物标志物,探讨其应用价值,进一步阐明IgAN的发病机制。方法:(1)入选年龄18～60岁,解放军总医院住院肾活检确诊为IgAN的患者10例,取活检肾组织进行研究;年龄18～60岁的于解放军总医院泌尿外科行肾癌肾脏切除的患者10例,取肾癌患者癌旁6cm以外的正常肾脏组织作为对照组进行研究。收集上述人群的临床基本信息及化验检查结果。上述人群肾脏组织利用Raybiotech细胞因子抗体芯片进行检测,筛选差异蛋白质,并对差异蛋白质进行GO(Gene Ontology)分析。结合分析结果,选取了 IL-33利用酶联免疫吸附试验(ELISA)进行验证。(2)对解放军总医院肾科肾穿刺活检患者进行筛选,入组20-60岁的48例诊断为IgANⅡ～Ⅲ级的患者和9例诊断为IMNⅠ～Ⅱ期的患者;另入组40例20-60岁尿常规、血常规及血生化结果无明显异常且无基础疾病的正常人作为健康对照。收集上述人群晨起第1次尿中段尿,通过Lable free LC/MS高分辨质谱技术对29例IgAN患者9例IMN患者和40例健康对照进行尿液蛋白质检测,筛选差异蛋白质,对差异蛋白质进行GO富集分析;依据蛋白质谱和数据分析的结果,结合IgAN可能的发病机制,选取48例IgAN患者和40例正常人尿液利用酶联免疫吸附试验(ELISA)对转铁蛋白(transferrin,TF)进行验证。结果:(1)对10例病理诊断为IgANⅡ~Ⅲ级、10例正常肾脏组织进行细胞因子表达谱检测,共发现了 25个差异蛋白质,GO分析结果表明:这些蛋白质主要与应激、细胞粘附、蛋白质代谢、信号转导相关。ELISA验证结果显示:IgAN患者肾组织中IL33表达水平显著高于对照组。(2)利用高分辨质谱技术检测发现,与正常对照组相比对,在IgAN和IMN疾病组,检测到53种蛋白质差异表达(p0.05);在IMN组中找到15种显著差异表达但在IgAN组中无差异表达的蛋白质(p0.05);在IgAN组中找到13种差异表达但在IMN组中无差异表达的蛋白质(p0.05)。IMN组和IgAN组组间,找到了 9种蛋白质差异表达(p0.05)。GO富集发现这些差异蛋白质主要参与凝血与血栓形成、炎症反应、补体系统激活、蛋白转运、铁代谢、糖及脂质代谢生物过程。差异蛋白C3、EGF、APOD等与以往研究结果相符,ORM1、ORM2可能是诊断IgAN和IMN的生物标志物。ELISA验证结果显示:IgAN患者尿液中TF表达水平显著高于正常对照组,TF具有非常大的诊断价值。结论:IL-33可能是IgAN发病机制中起重要作用的细胞因子,TF可能是优于24h尿蛋白定量诊断IgAN和IMN的早期生物标志物,ORM1、ORM2,尤其ORM2在IMN患者尿液中高表达可能是IMN区别于IgAN的重要标志物,CETP可能是诊断IgAN的生物标志物。
[Abstract]:Background and Objective: IgA nephropathy (IgAN) is one of the most common primary glomerular diseases in the world, which is one of the most important causes of end-stage renal failure. 30-40% of the IgAN patients developed end-stage renal disease (ESRD) in 20 years after the diagnosis of the disease. The heavy burden on the family and society. The diagnosis of IgAN is still dependent on renal biopsy, but the biopsy of the kidney is invasive and there is a certain risk and contraindication. The diagnosis and treatment of IgAN is very important for prevention and control and reduction of the number of ESRD patients. Therefore, it is of great significance to explore the pathogenesis of IgAN and the early invasive biomarkers. In this study, the renal tissue of the IgAN patients and the control group were tested by the technique of Raybitech's cytokine antibody chip. The detection and analysis of the urine of the patients with IgAN and the primary membranous nephropathy (IMN) and the normal population were analyzed by high-resolution mass spectrometry. The biological markers of the IgAN were selected. The application value of IgAN is discussed, and the pathogenesis of IgAN is further clarified. Methods: (1) 10 patients with IgAN were selected for the diagnosis of renal biopsy from 18 to 60 years old and in the general hospital of the PLA. The normal kidney tissues except 6cm in the carcinoma of the renal cell carcinoma were studied as the control group. Collect the clinical basic information and test results of the above-mentioned population. In the above-mentioned population, the renal tissue was tested using a Raybitech cytokine antibody chip to screen the differential protein, and a GO (Gene Ontology) analysis of the differential protein was performed. In combination with the results of the analysis, an enzyme-linked immunosorbent assay (ELISA) was used for the validation of IL-33. (2) The patients with the renal biopsy of the General Hospital of the PLA General Hospital were selected, 48 of the patients aged 20 to 60 were diagnosed as the patients with the IgAN 鈪，
本文编号：2362360
[Abstract]:Background and Objective: IgA nephropathy (IgAN) is one of the most common primary glomerular diseases in the world, which is one of the most important causes of end-stage renal failure. 30-40% of the IgAN patients developed end-stage renal disease (ESRD) in 20 years after the diagnosis of the disease. The heavy burden on the family and society. The diagnosis of IgAN is still dependent on renal biopsy, but the biopsy of the kidney is invasive and there is a certain risk and contraindication. The diagnosis and treatment of IgAN is very important for prevention and control and reduction of the number of ESRD patients. Therefore, it is of great significance to explore the pathogenesis of IgAN and the early invasive biomarkers. In this study, the renal tissue of the IgAN patients and the control group were tested by the technique of Raybitech's cytokine antibody chip. The detection and analysis of the urine of the patients with IgAN and the primary membranous nephropathy (IMN) and the normal population were analyzed by high-resolution mass spectrometry. The biological markers of the IgAN were selected. The application value of IgAN is discussed, and the pathogenesis of IgAN is further clarified. Methods: (1) 10 patients with IgAN were selected for the diagnosis of renal biopsy from 18 to 60 years old and in the general hospital of the PLA. The normal kidney tissues except 6cm in the carcinoma of the renal cell carcinoma were studied as the control group. Collect the clinical basic information and test results of the above-mentioned population. In the above-mentioned population, the renal tissue was tested using a Raybitech cytokine antibody chip to screen the differential protein, and a GO (Gene Ontology) analysis of the differential protein was performed. In combination with the results of the analysis, an enzyme-linked immunosorbent assay (ELISA) was used for the validation of IL-33. (2) The patients with the renal biopsy of the General Hospital of the PLA General Hospital were selected, 48 of the patients aged 20 to 60 were diagnosed as the patients with the IgAN 鈪，
本文编号：2362360
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