[GATA1在支气管哮喘中的表达及其对ORMDL3基因转录调控的影响]。

Q3 Medicine

中国当代儿科杂志 Pub Date : 2025-02-15 DOI:10.7499/j.issn.1008-8830.2408143

Hu Chen, Jiao-Jiao Li, Yue Yuan, Rui Jin

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引用次数: 0

摘要

目的：研究转录因子GATA1在支气管哮喘（简称哮喘）中的表达及其对哮喘易感基因orosomucoid 1-like protein 3 （ORMDL3）表达水平的影响及其潜在的分子机制。方法：从基因表达综合数据库（Gene Expression Omnibus， GEO）中选取28例中度哮喘、46例重度哮喘和12例正常人作为研究对象。分析哮喘患者与正常人GATA1、ORMDL3 mRNA表达水平及其相关性。将pGL-185/58质粒与GATA1基因siRNA （si-GATA1组）和siRNA阴性对照（si-对照组）共转染BEAS-2B细胞。采用生物信息学方法预测ORMDL3基因启动子区域GATA1结合位点。采用双荧光素酶报告基因系统评估ORMDL3启动子活性，采用实时定量PCR和Western blotting检测GATA1和ORMDL3 mRNA和蛋白表达水平。采用染色质免疫沉淀法（ChIP）测定GATA1是否与ORMDL3的启动子区结合。结果：重度哮喘组GATA1、ORMDL3 mRNA表达水平显著高于正常对照组(Pr分别为0.636、0.341；PORMDL3启动子荧光素酶活性以及ORMDL3 mRNA和蛋白的表达水平在si-GATA1组中低于si-对照组（PORMDL3）。结论：哮喘患者GATA1表达升高，可能调控哮喘易感基因ORMDL3启动子活性及表达。

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[Expression of GATA1 in bronchial asthma and its effect on the transcription regulation of the ORMDL3 gene].

Objectives: To study the expression of the transcription factor GATA1 in bronchial asthma (referred to as asthma) and its effect on the expression level of the asthma susceptibility gene orosomucoid 1-like protein 3 (ORMDL3), along with the underlying molecular mechanisms.

Methods: The study included 28 cases of moderate asthma, 46 cases of severe asthma, and 12 normal controls from the Gene Expression Omnibus (GEO) database. The mRNA expression levels of GATA1 and ORMDL3 were analyzed among the asthma patients and the normal controls, including their correlation. The pGL-185/58 plasmid was co-transfected with GATA1 gene siRNA (si-GATA1 group) and siRNA negative control (si-control group) into BEAS-2B cells. Bioinformatics methods were used to predict GATA1 binding sites in the promoter region of the ORMDL3 gene. The dual-luciferase reporter gene system was employed to assess the promoter activity of ORMDL3, while real-time quantitative PCR and Western blotting were used to measure the mRNA and protein expression levels of GATA1 and ORMDL3. Chromatin immunoprecipitation (ChIP) assays were conducted to determine whether GATA1 binds to the promoter region of ORMDL3.

Results: The expression levels of GATA1 and ORMDL3 mRNA were significantly higher in the severe asthma group compared to the normal control group (P<0.001). Positive correlations were observed between GATA1 mRNA and ORMDL3 mRNA expression levels in both the moderate and severe asthma groups (r=0.636 and 0.341, respectively; P<0.05). In BEAS-2B cells, the dual-luciferase reporter assay revealed that ORMDL3 promoter luciferase activity, as well as ORMDL3 mRNA and protein expression levels, were lower in the si-GATA1 group compared to the si-control group (P<0.05). ChIP assay results demonstrated that GATA1 could bind to the promoter region of ORMDL3.

Conclusions: The expression of GATA1 is increased in asthma patients, which may regulate the promoter activity and expression of the asthma susceptibility gene ORMDL3.

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来源期刊

中国当代儿科杂志 Medicine-Pediatrics, Perinatology and Child Health

CiteScore

1.50

自引率

0.00%

发文量

5006

期刊介绍： The Chinese Journal of Contemporary Pediatrics (CJCP) is a peer-reviewed open access periodical in the field of pediatrics that is sponsored by the Central South University/Xiangya Hospital of Central South University and under the auspices of the Ministry of Education of China. It is cited as a source in the scientific and technological papers of Chinese journals, the Chinese Science Citation Database (CSCD), and is one of the core Chinese periodicals in the Peking University Library. CJCP has been indexed by MEDLINE/PubMed/PMC of the American National Library, American Chemical Abstracts (CA), Holland Medical Abstracts (EM), Western Pacific Region Index Medicus (WPRIM), Scopus and EBSCO. It is a monthly periodical published on the 15th of every month, and is distributed both at home and overseas. The Chinese series publication number is CN 43-1301/R；ISSN 1008-8830. The tenet of CJCP is to “reflect the latest advances and be open to the world”. The periodical reports the most recent advances in the contemporary pediatric field. The majority of the readership is pediatric doctors and researchers.