Inhibition of matrix metalloproteinase MMP-2 activates chloride current in human airway epithelial cells.
Matrix metalloproteinases (MMPs) are involved in the remodeling and degradation of the extracellular matrix. Recently, it has been found that MMPs also contribute to processes not directly related to tissue remodeling, such as platelet aggregation or degranulation of airway gland cells. Since mucus secretion is closely related to ion channel function, we investigated whether MMPs could also be involved in the regulation of ion channels. We used human airway submucosal cell line Calu-3 to study the effects of MMPs on whole-cell current and transepithelial short-circuit current (I(sc)). Phenanthroline, a specific inhibitor of MMPs, increased whole-cell current with the half-maximally effective dose of 5.2 microM, and reversibly activated I(sc) in transepithelial measurements. Current stimulated by phenanthroline displayed linear current-voltage relationships and had inhibitor pharmacology and ion selectivity consistent with cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel activity. Zymography and Western blot showed significant expression of MMP-2 in Calu-3 cells. Moreover, anti-MMP-2 antibodies (1 microg/mL) increased whole-cell current and I(sc), whereas human recombinant MMP-2 (10 ng/mL) reduced it. We also studied the expression of MMPs and the effects of phenanthroline on whole-cell current in A549 cells, which are derived from airway surface epithelium and do not express CFTR Cl- channels. While these cells also showed significant expression of MMP-2, inhibition of this enzyme with phenanthroline exerted no significant effect on whole-cell current. It is concluded that MMP-2 is involved in the regulation of CFTR Cl- channels in human airways.
Duszyk
M
M
Department of Physiology, University of Alberta, Edmonton, Canada. mduszyk@gpu.srv.ualberta.ca
Shu
Y
Y
Sawicki
G
G
Radomski
A
A
Man
S F
SF
Radomski
M W
MW
D002460:Cell Line; D018118:Chloride Channels / Q000187:drug effects*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000502:physiology; D061965:Matrix Metalloproteinase Inhibitors; D018408:Patch-Clamp Techniques; D010618:Phenanthrolines / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D011650:Pulmonary Alveoli / Q000187:drug effects* / Q000201:enzymology
C505032:CFTR protein, human; D018118:Chloride Channels; D061965:Matrix Metalloproteinase Inhibitors; D010618:Phenanthrolines; D011480:Protease Inhibitors; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D020778:Matrix Metalloproteinase 2; C025205:1,10-phenanthroline