Steiger D, Fahy J, Boushey H, Finkbeiner W E, Basbaum C
Department of Anatomy, University of California, San Francisco 94143.
Am J Respir Cell Mol Biol. 1994 May;10(5):538-45. doi: 10.1165/ajrcmb.10.5.8179917.
Mucus hypersecretion is a prominent feature of the airway's response to injury. The ability to quantitatively detect mucin and mucin mRNA in vivo in human airways would facilitate the determination of safe exposure levels to various air pollutants and the identification of drugs capable of attenuating mucus hypersecretion. To this end, we have developed two assays: an enzyme-linked immunosorbent assay (ELISA) quantifying mucin-like molecules and a polymerase chain reaction (PCR)-based assay quantifying mucin mRNA. These tests are performed on bronchial lavage fluid and epithelial cells brushed from the surfaces of human airways at bronchoscopy. The PCR data are normalized to eliminate potentially confounding effects of nonepithelial cells in the samples. In a study of six smokers and six nonsmokers, the ELISA detected significantly more mucin-like material in the airways of the smokers than of the nonsmokers. The median mucin concentration for the smokers was 52.2 micrograms/ml (range, 16.3 to 4,860.0), whereas that for the nonsmokers was 12.7 micrograms/ml (range, 4.5 to 22.9). The difference between smokers and nonsmokers was statistically significant (P < or = 0.01). The PCR-based test showed a trend for RNA samples from smokers to be enriched (vis-à-vis nonsmokers) in mucin mRNA.
黏液高分泌是气道对损伤反应的一个突出特征。能够在体内定量检测人类气道中的黏蛋白和黏蛋白mRNA，将有助于确定对各种空气污染物的安全暴露水平，并有助于识别能够减轻黏液高分泌的药物。为此，我们开发了两种检测方法：一种是定量黏蛋白样分子的酶联免疫吸附测定（ELISA），另一种是定量黏蛋白mRNA的基于聚合酶链反应（PCR）的检测方法。这些检测在支气管灌洗液和支气管镜检查时从人类气道表面刷取的上皮细胞上进行。对PCR数据进行标准化处理，以消除样本中非上皮细胞可能产生的混杂影响。在一项对6名吸烟者和6名不吸烟者的研究中，ELISA检测到吸烟者气道中的黏蛋白样物质明显多于不吸烟者。吸烟者的黏蛋白浓度中位数为52.2微克/毫升（范围为16.3至4860.0），而不吸烟者的为12.7微克/毫升（范围为4.5至22.9）。吸烟者和不吸烟者之间的差异具有统计学意义（P≤0.01）。基于PCR的检测显示，吸烟者的RNA样本中黏蛋白mRNA相对于不吸烟者有富集的趋势。
