Sanghavi J N, Rabe K F, Kim J S, Magnussen H, Leff A R, White S R
Department of Medicine, University of Chicago, Illinois 60637.
Am J Respir Cell Mol Biol. 1994 Aug;11(2):181-7. doi: 10.1165/ajrcmb.11.2.8049078.
Calcitonin gene-related peptide (CGRP) is contained within and secreted by nerves and neuroepithelial bodies in the airway epithelium. To determine whether CGRP is a chemoattractant, tracheal epithelial cells isolated from 23 guinea pigs, and bronchial epithelial cells isolated from seven human donors were grown in primary culture for 4 to 5 days. Cell migration was assessed in a blindwell chemotaxis chamber. A gelatin-coated polycarbonate filter (8-microns pore size) separated the upper wells containing 5 x 10(4) cells from the lower wells containing chemoattractant (either CGRP or insulin). Cells were stimulated for 6 h, after which migrated cells on the filter were stained and counted. Both insulin and CGRP elicited migration of guinea pig tracheal epithelial (GPTE) cells. Exposure to 30 micrograms/ml insulin caused migration of 26.5 +/- 4.0 cells versus 4.3 +/- 0.6 cells per 10 hpf for control (P < 0.001). Treatment with 10(-9) M CGRP elicited migration of 39.6 +/- 2.5 cells versus 2.6 +/- 0.9 cells per 10 hpf for control (P < 0.001). Human bronchial epithelial (HBE) cells also migrated after stimulation with either insulin or CGRP: after 10(-9) M CGRP, HBE cell migration was 41.0 +/- 6.1 cells versus 3.4 +/- 0.3 cells per 10 hpf (P < 0.002; n = 3). Checkerboard analysis showed that the migration to CGRP was both chemotactic and chemokinetic. Incubation with 10(-8) M hCGRP-(8-37), a receptor antagonist for CGRP, plus 10(-9) M CGRP decreased GPTE cell migration to 9.8 +/- 2.4 cells versus 38.6 +/- 1.2 cells per 10 hpf for 10(-9) M CGRP alone (P < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)
降钙素基因相关肽（CGRP）存在于气道上皮的神经和神经上皮小体中并由其分泌。为了确定CGRP是否为一种趋化因子，从23只豚鼠分离出的气管上皮细胞以及从7名人类供体分离出的支气管上皮细胞进行原代培养4至5天。在盲孔趋化室中评估细胞迁移。用明胶包被的聚碳酸酯滤膜（孔径8微米）将含有5×10⁴个细胞的上孔与含有趋化因子（CGRP或胰岛素）的下孔隔开。细胞被刺激6小时，之后对滤膜上迁移的细胞进行染色并计数。胰岛素和CGRP均能引起豚鼠气管上皮（GPTE）细胞的迁移。暴露于30微克/毫升胰岛素时，每10高倍视野（hpf）迁移的细胞数为26.5±4.0个，而对照组为4.3±0.6个（P<0.001）。用10⁻⁹M CGRP处理时，每10 hpf迁移的细胞数为39.6±2.5个，而对照组为2.6±0.9个（P<0.001）。人支气管上皮（HBE）细胞在用胰岛素或CGRP刺激后也会迁移：在10⁻⁹M CGRP作用后，HBE细胞每10 hpf的迁移数为41.0±6.1个，而对照组为3.4±0.3个（P<0.002；n = 3）。棋盘分析表明，向CGRP的迁移既是趋化性的也是化学动力学性的。用10⁻⁸M hCGRP -（8 - 37）（一种CGRP受体拮抗剂）与10⁻⁹M CGRP共同孵育，使GPTE细胞迁移数降至每10 hpf 9.8±2.4个，而单独使用10⁻⁹M CGRP时为38.6±1.2个（P<0.0001）。（摘要截短于250字）
