TY - JOUR
T1 - Antiproteases attenuate the release of neutrophil chemotactic activity from bronchial epithelial cells induced by smoke
AU - Koyama, Sekiya
AU - Rennard, Stephen I.
AU - Leikauf, George D.
AU - Ertl, Ronald F.
AU - Robbins, Richard A.
PY - 1996
Y1 - 1996
N2 - 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 α-1-protease inhibitor (α-1-PI), chloromethyl ketone (CK) derivatives. N-tosyl-L-lysine CK (TLCK), methylsuccinyl-Ala-Ala-Pro-Val CK (SPCK). N-α-tosyl-L- phenylalanine CK (TPCK), and N-α-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.
AB - 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 α-1-protease inhibitor (α-1-PI), chloromethyl ketone (CK) derivatives. N-tosyl-L-lysine CK (TLCK), methylsuccinyl-Ala-Ala-Pro-Val CK (SPCK). N-α-tosyl-L- phenylalanine CK (TPCK), and N-α-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.
KW - bronchial epithelial cells
KW - neutrophil chemotactic activity
KW - serine protease inhibitor
KW - smoking
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U2 - 10.3109/01902149609074014
DO - 10.3109/01902149609074014
M3 - Article
C2 - 8838132
AN - SCOPUS:0030052644
SN - 0190-2148
VL - 22
SP - 1
EP - 19
JO - Experimental Lung Research
JF - Experimental Lung Research
IS - 1
ER -