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ANALYSIS OF MACROPHAGE SPECIFICalpha1-ANTITRYPSIN PROMOTER IN PATIENTS WITH EMPHYSEMA

肺气肿患者巨噬细胞特异性α1-抗胰蛋白酶启动子的分析

基本信息

批准号：
07670664
负责人：
HASEGAWA Yoshinori
金额：
$ 1.54万
依托单位：
NAGOYA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670664/
关键词：
EMPHYSEMA macrophage specific promoter alpha1-antitrypsin gene α_1-アンチトリプシン遺伝子

项目摘要

The purpose of this study is to investigate the genetic factors for pulmonary emphysema. We analyzed the genetic polymorphism of macrophage specific promoter sequences of alpha1-antitrypsin gene. Mononuclear cells were separated from peripheral blood of patients with pulmonary emphysema, chronic bronchiolitis and bronchial asthma, and the genomic DNA was isolated. Macrophage specific promoter sequences of alpha1-antitrypsin gene was amplified by polymerase chain reaction. Single strand conformational polymorphism analysis was adopted on the analysis of the amplified DNA sequences. We detected three types of genetic polymorphism of macrophage specific promoter sequences of alpha1-antitrypsin gene by single-strand conformational polymorphism analysis. The genetic polymorphism was not associated with the diseases which were investigated. Then, we cloned the cDNA of macrophage specific promoter sequences of alpha1-antitrypsin gene from the normal subjects. The sequences were inserted into pGL2-basic vector, which contains luciferase gene as a reporter gene to test the unknown promoter activity. Our results demonstrated that macrophage specific promoter sequences of alpha1-antitrypsin gene showed the weak promoter activity itself, and the macrophage specific promoter activity was not clear. These findings suggest that further investigation would be necessary to study the high activity and macrophage specificity of the promoter sequences for alpha1-antitrypsin gene after stimulation of the gene transfected cells by cytokines or other factors.

本研究的目的是探讨肺气肿的遗传因素。我们分析了α 1-抗胰蛋白酶基因巨噬细胞特异性启动子序列的遗传多态性。从肺气肿、慢性毛细支气管炎和支气管哮喘患者外周血中分离单核细胞，提取基因组DNA。用聚合酶链反应扩增巨噬细胞特异性α 1-抗胰蛋白酶基因启动子序列。采用单链构象多态性分析对扩增的DNA序列进行分析。应用单链构象多态性分析方法检测了巨噬细胞特异性α 1-抗胰蛋白酶基因启动子序列的3种遗传多态性。该基因多态性与所调查的疾病无关。然后，我们克隆了正常人巨噬细胞特异性α 1-抗胰蛋白酶基因启动子序列的cDNA。将该序列插入到含有荧光素酶基因作为报告基因的pGL 2-basic载体中，以测试未知的启动子活性。结果表明，α 1-抗胰蛋白酶基因的巨噬细胞特异性启动子序列本身具有较弱的启动子活性，而巨噬细胞特异性启动子活性不明显。这些结果表明，有必要进一步研究α 1-抗胰蛋白酶基因的启动子序列的高活性和巨噬细胞特异性的基因转染细胞的细胞因子或其他因素的刺激后。