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聚合酶链反应扩增人支气管肺泡灌洗细胞中生长因子的信使核糖核酸。

Polymerase chain reaction amplification of messages for growth factors in cells from human bronchoalveolar lavage fluids.

作者信息

Adolff C H, Golden J A, Kennedy P W, Goetzl E J, Turck C W

机构信息

Department of Medicine, University of California Medical Center, San Francisco.

出版信息

Inflammation. 1991 Aug;15(4):259-68. doi: 10.1007/BF00917311.

Abstract

Genetic messages for polypeptide growth factors were assessed in human alveolar macrophages, obtained by bronchoalveolar lavage (BAL) from normal subjects (N = 3) and from patients with pneumonia (N = 3), pulmonary lymphoma (N = 3), and idiopathic pulmonary fibrosis (N = 3). Complementary DNAs (cDNAs) were prepared by reverse transcription of the RNA extracted from alveolar macrophages before and after culture on a plastic surface. The cDNAs encoding 10 different growth factors were amplified for electrophoretic analysis by polymerase chain reaction with a pair of 3' and 5' primers specific for each factor. Alveolar macrophages from all normal subjects and patients expressed the messages for interleukin-1 beta and transforming growth factor-beta. Alveolar macrophages from some normal subjects also contained message for insulin-like growth factor-1. Alveolar macrophages from six of nine patients with lung diseases also expressed messages for one or more additional growth factors, including epidermal growth factor, transforming growth factor-alpha, interleukin-1 alpha, and platelet-derived growth factor. The polymerase chain reaction technique thus permits determination of the profile of growth factors contributed to pulmonary reactions by alveolar macrophages, which may be important in pulmonary healing and fibrosis.

摘要

通过支气管肺泡灌洗(BAL)从正常受试者(N = 3)以及患有肺炎(N = 3)、肺淋巴瘤(N = 3)和特发性肺纤维化(N = 3)的患者中获取人肺泡巨噬细胞,评估其中多肽生长因子的遗传信息。在塑料表面培养前后,从肺泡巨噬细胞中提取RNA,通过逆转录制备互补DNA(cDNA)。使用针对每种因子的一对3'和5'引物,通过聚合酶链反应扩增编码10种不同生长因子的cDNA,用于电泳分析。所有正常受试者和患者的肺泡巨噬细胞均表达白细胞介素-1β和转化生长因子-β的信息。一些正常受试者的肺泡巨噬细胞还含有胰岛素样生长因子-1的信息。9名肺部疾病患者中有6名的肺泡巨噬细胞还表达了一种或多种其他生长因子的信息,包括表皮生长因子、转化生长因子-α、白细胞介素-1α和血小板衍生生长因子。聚合酶链反应技术因此能够确定肺泡巨噬细胞对肺部反应所贡献的生长因子谱,这在肺部愈合和纤维化过程中可能很重要。

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