PPACK attenuates plasmin-induced changes in endothelial integrity.
In order to determine whether plasmin affects endothelial cell integrity directly, confluent bovine aortic endothelial cells were treated with plasminogen and a plasminogen activator. The permeability of the monolayer to [125I]-albumin was shown to be increased significantly (P < 0.01) with a concomitant decrease in viability. Plasmin activity correlated significantly with endothelial cell permeability (p < 0.004; r = 0.82). Coincubation with D-phenylalanyl-L-prolyl-L-arginyl chloromethylketone, a tripeptide inhibitor of plasmin, reduced the increase in endothelial permeability induced by plasmin by 59% (p = 0.033). Monolayers studied in parallel were stained with rhodamine-phalloidin to visualize F-actin. There were significant morphologic changes in the endothelial monolayers exposed to plasmin compared to control monolayers, and these changes could be attenuated by coincubation with D-phenylalanyl-L-prolyl-L-arginyl chloromethylketone. These studies show that: 1) plasmin induces significant increases in endothelial cell permeability with accompanying morphologic changes; and 2) these deleterious functional and morphologic effects can be attenuated by coincubation with the plasmin inhibitor, D-phenylalanyl-L-prolyl-L-arginyl chloromethylketone.
Rabbani
L E
LE
Department of Medicine, Brigham and Women's Hospital, Boston, MA.
Johnstone
M T
MT
Rudd
M A
MA
Devine
P
P
George
D
D
Loscalzo
J
J
D000199:Actins / Q000032:analysis; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D000933:Antifibrinolytic Agents / Q000494:pharmacology*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D005341:Fibrinolysin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D008969:Molecular Sequence Data
D000199:Actins; D000590:Amino Acid Chloromethyl Ketones; D000933:Antifibrinolytic Agents; D005343:Fibrinolytic Agents; D005341:Fibrinolysin; C040308:phenylalanyl-prolyl-arginine-chloromethyl ketone