title: Antiproteases attenuate the release of neutrophil chemotactic activity from bronchial epithelial cells induced by smoke.
abstract: The released neutrophil chemotactic activity (NCA) from bronchial epithelial cells (BECs) in response to smoke extract was evaluated by reverse-phase, high-performance liquid chromatography (RP-HPLC) and the involvement of proteolytic activity was assessed for the release of NCA from BECs. Smoke extract stimulated the release of NCA (55.3 +/- 5.2 vs. 17.3 +/- 4.1 cells per high-power field [HPF], p < .001). The released activity determined by RP-HPLC analysis was 15-hydroxyeicosatetraenoic acid and leukotriene B4. Several structurally and functionally different serine protease inhibitors, including alpha-1-protease inhibitor (alpha-1-PI), chloromethyl ketone (CK) derivatives, N-tosyl-L-lysine CK (TLCK), methoxysuccinyl-Ala-Ala-Pro-Val CK (SPCK), N-alpha-tosyl-L-phenylalanine CK (TPCK), and N-alpha-p-tosyl-L-arginine methyl ester hydrochloride (TAME), attenuated the release of NCA (P < .01) in a dose-dependent fashion. Leupeptin, a cysteine protease inhibitor, has only a small effect on the release of NCA (p < .05), and phosphoramidon, a neutral endopeptidase inhibitor, had no effect. The measurement of proteolytic enzyme activity using synthetic substrate S-2288 revealed that smoke extract significantly (p < .05) augmented the serine protease activity in BEC layers. Culture supernatant fluids and cell lysates of BECs in response to smoke extract solubilized 14C-labeled casein. These results suggest that BECs may release lipoxygenase-derived NCA in response to smoke extract and that the release of NCA may involve the activation of proteolytic activity of BECs which was inhibited by serine protease inhibitors.
authors.affiliation: First Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Koyama
authors.forename: S I
authors.initials: SI
authors.lastname: Rennard
authors.forename: G D
authors.initials: GD
authors.lastname: Leikauf
authors.forename: R F
authors.initials: RF
authors.lastname: Ertl
authors.forename: R A
authors.initials: RA
authors.lastname: Robbins
mesh_terms: D000418:Albumins / Q000494:pharmacology; D000818:Animals; D001980:Bronchi / Q000187:drug effects*; D002417:Cattle; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000378:metabolism*; D002633:Chemotaxis / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000378:metabolism; D004848:Epithelium / Q000187:drug effects; D006020:Glycopeptides / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D012697:Serine Endopeptidases / Q000378:metabolism*; D012906:Smoke / Q000009:adverse effects*; D014105:Tosyl Compounds / Q000494:pharmacology
chemical_list: D000418:Albumins; D002630:Chemotactic Factors; D015777:Eicosanoids; D006020:Glycopeptides; D007976:Leupeptins; D012906:Smoke; D014105:Tosyl Compounds; D012697:Serine Endopeptidases; C032854:leupeptin; C008890:phosphoramidon