本文選題：特發(fā)性肺纖維化 + 基質(zhì)金屬蛋白酶-1　； 參考：《河北醫(yī)科大學(xué)》2012年碩士論文

【摘要】：特發(fā)性肺纖維化是一種慢性致命性、以肺部組織異常重塑為特征的間質(zhì)性肺部疾病。其病因不明，目前認(rèn)為是環(huán)境因素與多種遺傳因素相互作用可能參與其疾病的發(fā)展。基質(zhì)金屬蛋白酶-1其主要作用是降解細(xì)胞外基質(zhì)成份，認(rèn)為其在肺纖維化的形成中起了重要作用。近年來(lái)研究顯示MMP-1具有基因多態(tài)性現(xiàn)象，其與哮喘、慢性阻塞型肺疾病、肺結(jié)核等多種肺疾病之間的易感性已有報(bào)道。而國(guó)內(nèi)尚無(wú)關(guān)于MMP-1基因多態(tài)性與特發(fā)性肺纖維化的研究報(bào)道。本實(shí)驗(yàn)旨在通過(guò)研究基質(zhì)金屬蛋白酶-1（Matrix metalloproteinase-1）啟動(dòng)子區(qū)-1607位點(diǎn)1G/2G單核苷酸基因多態(tài)性(single nucleotide polymorphism，，SNP）與中國(guó)河北地區(qū)漢族特發(fā)性肺纖維化的相關(guān)性，并探測(cè)特發(fā)性肺纖維化患者血清學(xué)MMP-1和TIMP-1的蛋白含量，探討中國(guó)河北地區(qū)漢族特發(fā)性肺纖維化患者的遺傳易感因素、發(fā)病機(jī)制及該位點(diǎn)基因多態(tài)性在不同種族特發(fā)性肺纖維化患者易感性方面的差異。 方法：采用基于人群的病例-對(duì)照研究方法，參考2002年中華醫(yī)學(xué)會(huì)頒布的《特發(fā)性肺纖維化診斷和治療指南（草案）》中的診斷標(biāo)準(zhǔn)，選取臨床上診斷為特發(fā)性肺纖維化（IPF）漢族患者84例，其中男性51例，女性33例，年齡43歲-88歲，平均（66.7±9.5）歲，且除外繼發(fā)性肺纖維化、急性感染、腫瘤、栓塞、心衰、控制不良的糖尿病、消化性潰瘍、嚴(yán)重骨質(zhì)疏松、結(jié)核病患者。對(duì)照組：選取漢族健康體檢人群100例，其中男性53例，女性47例，年齡40歲-73歲，平均年齡（50.2±7.5）歲，且既往無(wú)變態(tài)反應(yīng)性疾病病史，近期無(wú)感染，無(wú)免疫抑制劑應(yīng)用史。采集IPF患者及健康對(duì)照組外周靜脈血5ml，其中2mlEDTA抗凝，以消化-苯酚-氯仿有機(jī)溶劑法和樹(shù)脂型TM基因組DNA提純?cè)噭┖刑崛⊥庵苎准?xì)胞DNA，利用限制性?xún)?nèi)切酶ALUI，聚合酶鏈反應(yīng)-限制性片段長(zhǎng)度多態(tài)性(PCR-RFLP）技術(shù)結(jié)合瓊脂糖凝膠電泳的方法，檢測(cè)IPF患者和健康對(duì)照者M(jìn)MP-1基因啟動(dòng)子區(qū)-1607(1G/2G)多態(tài)性位點(diǎn)的基因型和等位基因分布，分析兩組人群MMP-l基因型和等位基因頻率的差異。余3ml靜脈血促凝劑促凝，采用雙抗夾心酶聯(lián)免疫吸附法（ELISA）測(cè)定IPF患者和健康對(duì)照組MMP-1和TIMP-1的血清蛋白表達(dá)水平。 結(jié)果： 1MMP-1位點(diǎn)基因多態(tài)性分析 1.1MMP-1-1607的基因型 本實(shí)驗(yàn)檢測(cè)得到三種基因型：1G/1G型、1G/2G型及2G/2G型；其中1G/1G型為野生型，1G/2G型及2G/2G型為突變型。IPF組與健康對(duì)照組均符合Hardy-Weinberg遺傳平衡性檢驗(yàn)（P0.05），兩組之間具有良好的可比性。 1.2MMP-1-16071G/2G各基因型在兩組間的分布情況 IPF組84例中檢測(cè)出1G/1G基因型12例(14.3%)，1G/2G基因型30例(35.7%)，2G/2G基因型42例(50%)。健康對(duì)照組100例中檢測(cè)出1G/1G基因型11例(11%)，1G/2G基因型42例(42%)，2G/2G基因型47例(47%)。MMP-1-1607位點(diǎn)各基因型在兩組間的分布頻率比較無(wú)明顯統(tǒng)計(jì)學(xué)差異（χ2=0.940, P0.05）。 1.3MMP-1-1607位點(diǎn)1G/2G各等位基因頻率在兩組間的分布情況 IPF組84例中1G等位基因頻率為32.1%，2G等位基因頻率為67.9%，健康對(duì)照組100例中1G等位基因頻率為32%，2G等位基因頻率為68%。MMP-1-1607位點(diǎn)的各等位基因在兩組間的分布頻率比較無(wú)顯著統(tǒng)計(jì)學(xué)差異性（χ2=0.001, P0.05）。 2血清MMP-1，TIMP-1蛋白濃度的變化 2.1血清MMP-1蛋白濃度變化： IPF患者與健康對(duì)照組MMP-1血清蛋白濃度分別為：374.66±54.18ng/ml，97.45±20.39ng/ml。IPF患者血清MMP-1蛋白濃度明顯高于健康對(duì)照組，兩組比較有統(tǒng)計(jì)學(xué)意義（P0.05）。 2.2血清TIMP-1蛋白濃度變化： IPF組與健康對(duì)照組TIMP-1血清蛋白濃度分別為：111.10±26.70ng/ml，40.75±11.46ng/ml。IPF患者血清TIMP-1蛋白濃度較健康對(duì)照組明顯升高，兩組比較有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.3用MMP-1/TIMP-1蛋白濃度得出兩者的比值 IPF組與健康對(duì)照組MMP-1/TIMP-1比值分別為：3.39±1.06，2.72±1.00，兩組比較有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論： 1．結(jié)合本研究推測(cè)MMP-1-1607位點(diǎn)的1G/2G基因多態(tài)性與中國(guó)河北地區(qū)漢族人群特發(fā)性肺間質(zhì)纖維化的易感性無(wú)明顯相關(guān)性，結(jié)合對(duì)其它地區(qū)、種族人群的研究，說(shuō)明該位點(diǎn)基因多態(tài)性與IPF的易感性存在種族、地區(qū)的差異。 2. IPF患者肺纖維化的發(fā)生與MMP-1,TIMP-1水平升高可能有關(guān)，MMP-1和TIMP-1可能參與IPF疾病的發(fā)生發(fā)展。
[Abstract]:Idiopathic pulmonary fibrosis is a chronic and fatal interstitial lung disease characterized by abnormal remodeling of the lung tissue. Its etiology is unknown. It is considered that environmental factors interact with a variety of genetic factors and may participate in the development of its disease. Matrix metalloproteinase -1 is mainly used to degrade extracellular matrix components and consider it in the lung. There has been an important role in the formation of fibrosis. In recent years, studies have shown that MMP-1 has genetic polymorphisms, and its susceptibility to multiple lung diseases such as asthma, chronic obstructive pulmonary disease, tuberculosis and other lung diseases has been reported. However, there is no report on the polymorphism of MMP-1 gene and idiopathic pulmonary fibrosis in China. The aim of this experiment is to study the study. The correlation between 1G/2G single nucleotide polymorphism (single nucleotide polymorphism, SNP) in the promoter region of the matrix metalloproteinase -1 (Matrix metalloproteinase-1) and the Han idiopathic pulmonary fibrosis in Hebei, China, and detection of the protein content of MMP-1 and TIMP-1 in the blood of patients with idiopathic pulmonary fibrosis and the exploration of Hebei land in China Genetic susceptibility factors, pathogenesis and genetic polymorphisms of this loci in patients with idiopathic pulmonary fibrosis in different ethnic groups in patients with idiopathic pulmonary fibrosis.
Methods: a population based case control study was used, referring to the diagnostic criteria for the diagnosis and treatment of idiopathic pulmonary fibrosis (Draft) issued by the Chinese Medical Association of 2002, and 84 cases of Han patients diagnosed as idiopathic pulmonary fibrosis (IPF) were selected clinically, including 51 male, 33 female, 43 years old, and an average of (66.7 + 9.5). With the exception of secondary pulmonary fibrosis, acute infection, tumor, embolism, heart failure, poor control of diabetes, peptic ulcers, severe osteoporosis, and tuberculosis patients. Control group: 100 cases of Han health examination were selected, of which 53 were male, 47 women, age 40 -73 years, average age (50.2 + 7.5) years, and no previous allergy disease. History of disease, no infection, no history of immunosuppressant application. Collection of peripheral venous blood 5ml in IPF patients and healthy control group, including 2mlEDTA anticoagulant, extraction of DNA from peripheral blood leukocyte by digestion phenol chloroform organic solvent method and resin type TM genomic DNA purification kit, using restrictive endonuclease ALUI, polymerase chain reaction restriction fragment length PCR-RFLP technique combined with agarose gel electrophoresis to detect the genotype and allele distribution of the -1607 (1G/2G) polymorphic loci in the promoter region of the MMP-1 gene of IPF and healthy controls. The difference between the MMP-l genotypes and allele frequencies of the two groups was analyzed. The coagulant promoting coagulant in the venous blood of the two groups was used and the double resistant sandwich enzyme was used. Serum protein levels of MMP-1 and TIMP-1 in IPF patients and healthy controls were measured by ELISA.
Result錛

本文編號(hào)：1925186