Involvement of proteinase-activated receptor-2 in mast cell tryptase-induced barrier dysfunction in bovine aortic endothelial cells.
We report here a direct modulation by mast cell tryptase of endothelial barrier function through activation of proteinase-activated receptor-2 (PAR-2). In cultured bovine aortic endothelial cells (BAECs), tryptase, trypsin and PAR-2 activating peptide impaired the barrier function as determined by the permeability of protein-conjugated Evans blue. The tryptase-induced barrier dysfunction was completely blocked by U73122, and partially reversed by xestospongin C, calphostin C or Y27632. The intracellular Ca(2+) was elevated by tryptase. It was notable that ioxaglate, a contrast material that degranulates mast cells, markedly increased the permeability when applied to BAECs in combination with mast cells, an action that was blocked by nafamostat, a potent tryptase inhibitor. Immunofluorescence analysis showed that actin stress fibre formation and disruption of VE-cadherin were observed after exposure to tryptase or ioxaglate in combination with mast cells. Therefore, it is suggested that mast cell tryptase impairs endothelial barrier function through activation of endothelial PAR-2 in a manner dependent on the phospholipase C activity.
Sendo
Toshiaki
T
Department of Hospital Pharmacy, Faculty of Medicine, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Sumimura
Tomoko
T
Itoh
Yoshinori
Y
Goromaru
Takeshi
T
Aki
Keisei
K
Yano
Takahisa
T
Oike
Masahiro
M
Ito
Yushi
Y
Mori
Shuji
S
Nishibori
Masahiro
M
Oishi
Ryozo
R
D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000707:Anaphylaxis / Q000139:chemically induced / Q000378:metabolism / Q000503:physiopathology; D000818:Animals; D015703:Antigens, CD; D001011:Aorta / Q000166:cytology; D001550:Benzamidines; D015820:Cadherins / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002199:Capillary Permeability / Q000187:drug effects* / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D003287:Contrast Media / Q000009:adverse effects; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005070:Evans Blue; D016670:Gabexate / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D007485:Ioxaglic Acid / Q000494:pharmacology; D044464:Receptor, PAR-2 / Q000187:drug effects / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism* / Q000494:pharmacology; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D053802:Tryptases; D000099152:Cadherin 5
D000199:Actins; D015703:Antigens, CD; D001550:Benzamidines; D015820:Cadherins; D003287:Contrast Media; D004791:Enzyme Inhibitors; D005070:Evans Blue; D016670:Gabexate; D006146:Guanidines; D007485:Ioxaglic Acid; D044464:Receptor, PAR-2; D012697:Serine Endopeptidases; D053802:Tryptases; D000099152:Cadherin 5; C032855:nafamostat