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人呼吸道黏膜下腺细胞的表面活性物质蛋白A2基因表达

Surfactant protein A2 gene expression by human airway submucosal gland cells.

作者信息

Saitoh H, Okayama H, Shimura S, Fushimi T, Masuda T, Shirato K

机构信息

First Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Japan.

出版信息

Am J Respir Cell Mol Biol. 1998 Aug;19(2):202-9. doi: 10.1165/ajrcmb.19.2.3239.

Abstract

To determine whether human airway submucosal glands produce and secrete surfactant proteins, we examined their protein and gene expression in submucosal glands from trachea and bronchi obtained from operated and autopsied lungs within 4 h of death. Using a monoclonal antibody (PE-10) against surfactant protein A (SP-A), a positive immunoperoxidase stain was observed over serous cells of submucosal glands in histologic sections of airway walls. Measurement of SP-A in culture medium samples using single-step enzyme-linked immunosorbent assay showed a significant secretion of SP-A by isolated submucosal glands (1.2 +/- 0.08 ng/ml/h, SEM, n = 40). In gene expression experiments by reverse transciption-polymerase chain reaction, the SP-A complementary DNA (cDNA) segment was amplified from isolated submucosal glands, indicating the presence of SP-A messenger RNA (mRNA) in airway submucosal glands. Bronchial superficial epithelial cells failed to show the presence of SP-A mRNA. No cDNA segment of SP-B, SP-C, or SP-D cDNA was amplified from isolated submucosal glands or superficial epithelial cells, whereas all were amplified from alveolar tissue. Furthermore, in contrast to the control alveolar tissue, which expressed both SP-A1 and SP-A2 genes, SP-A2 gene transcript alone was detected in isolated submucosal glands by Southern analysis that included the digestion of the amplified SP-A cDNA fragment with the restriction enzyme Apa I. These findings indicate that human airway submucosal gland cells can transcribe the SP-A2 gene and produce SP-A protein in a manner different from peripheral airways and alveoli, playing a role in the airway defense mechanism.

摘要

为了确定人类气道黏膜下腺是否产生和分泌表面活性物质蛋白,我们在死亡后4小时内,对取自手术切除肺和尸检肺的气管及支气管黏膜下腺的蛋白和基因表达进行了检测。使用抗表面活性物质蛋白A(SP-A)的单克隆抗体(PE-10),在气道壁组织学切片的黏膜下腺浆液细胞上观察到阳性免疫过氧化物酶染色。使用单步酶联免疫吸附测定法测量培养基样本中的SP-A,结果显示分离出的黏膜下腺可大量分泌SP-A(1.2±0.08 ng/ml/h,标准误,n = 40)。在通过逆转录-聚合酶链反应进行的基因表达实验中,从分离出的黏膜下腺中扩增出了SP-A互补DNA(cDNA)片段,表明气道黏膜下腺中存在SP-A信使核糖核酸(mRNA)。支气管表面上皮细胞未显示出SP-A mRNA的存在。从分离出的黏膜下腺或表面上皮细胞中未扩增出SP-B、SP-C或SP-D cDNA片段,而从肺泡组织中均扩增出了这些片段。此外,与表达SP-A1和SP-A2基因的对照肺泡组织不同,通过Southern分析(包括用限制性内切酶Apa I消化扩增的SP-A cDNA片段)在分离出的黏膜下腺中仅检测到SP-A2基因转录本。这些发现表明,人类气道黏膜下腺细胞能够转录SP-A2基因并产生SP-A蛋白,其方式不同于外周气道和肺泡,在气道防御机制中发挥作用。

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