Broad-spectrum and selective serine protease inhibitors prevent expression of platelet-derived growth factor-BB and cerebral vasospasm after subarachnoid hemorrhage: vasospasm caused by cisternal injection of recombinant platelet-derived growth factor-BB.
OBJECTIVE
Plasma serine protease cascade, including the complement system and thrombin, is activated in the subarachnoid space during the acute phase after subarachnoid hemorrhage (SAH). To examine the effect of protease cascade-based inflammation and subsequent vascular repair in the development of cerebral vasospasm, we examined the effect of 2 synthetic serine protease inhibitors-FUT-175, an inhibitor of thrombin and the complement system, and argatroban, a selective inhibitor of thrombin-on the development of cerebral vasospasm in a rabbit SAH model.


METHODS
One hundred Japanese White male rabbits were used in the study. The SAH was simulated by a single injection of autologous arterial blood into the cisterna magna. To evaluate the development of cerebral vasospasm, the caliber of the basilar artery was measured on x-ray film before and at 2 days after SAH. Nine groups of rabbits (n=6 each) were treated with continuous intravenous injection of FUT-175 (2.5, 5, 10, or 20 mg/d), argatroban (1.25, 2.5, or 5 mg/d), or the same amount of saline (vehicle) for 48 hours, starting 40 minutes after SAH. Two days after SAH, the expression of homodimer of platelet-derived growth factor-BB (PDGF-BB) in the basilar artery was examined with immunohistochemical techniques. In 20 normal rabbits, 5 microg of recombinant PDGF-BB or vehicle was injected into the cisterna magna, and the basilar arteries were examined on angiograms for 48 hours.


RESULTS
Significant differences were observed in the caliber of the basilar arteries between the vehicle group and the groups with the 3 larger doses of FUT-175 (vehicle, 52+/-5.0%; 5 mg, 79+/-5.7%; 10 mg, 80+/-2.5%; 20 mg, 80+/-3.7%) and between the vehicle group and the groups with the 2 larger doses of argatroban (vehicle, 52+/-6.4%; 2.5 mg, 81+/-9.0%; 5 mg, 85+/-4.1%) (P<0.05). In the histological examination, administration of effective doses of FUT-175 or argatroban suppressed the expression of PDGF-BB in the endothelial and medial smooth muscle cell layers. Exogenous PDGF-BB caused delayed and prolonged vasoconstriction on normal basilar arteries.


CONCLUSIONS
Activation of the serine protease cascade and/or thrombin after SAH was demonstrated to play an essential role in the development of cerebral vasospasm. The expression of PDGF-BB-like protein in the arterial walls correlated with the development of cerebral vasospasm. Elevated PDGF-BB level in the subarachnoid space was found to induce delayed and chronic vasoconstriction.
Zhang
Z
Z
Laboratory for Cerebrovascular Disorders, Research Institute of National Cardiovascular Center, Osaka, Japan.
Nagata
I
I
Kikuchi
H
H
Xue
J H
JH
Sakai
N
N
Sakai
H
H
Yanamoto
H
H
D000818:Animals; D000991:Antithrombins / Q000494:pharmacology* / Q000627:therapeutic use; D001120:Arginine / Q000031:analogs & derivatives; D000077214:Becaplermin; D001550:Benzamidines; D002536:Cerebral Arteries / Q000000981:diagnostic imaging / Q000378:metabolism / Q000473:pathology; D002946:Cisterna Magna; D006146:Guanidines / Q000494:pharmacology* / Q000627:therapeutic use; D007150:Immunohistochemistry; D007267:Injections; D007700:Kinetics; D008297:Male; D010875:Pipecolic Acids / Q000494:pharmacology* / Q000627:therapeutic use; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis* / Q000276:immunology / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D011817:Rabbits; D011859:Radiography; D011994:Recombinant Proteins / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D013345:Subarachnoid Hemorrhage / Q000150:complications*; D013449:Sulfonamides; D020301:Vasospasm, Intracranial / Q000209:etiology* / Q000378:metabolism / Q000517:prevention & control
D000991:Antithrombins; D001550:Benzamidines; D006146:Guanidines; D010875:Pipecolic Acids; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D013449:Sulfonamides; D000077214:Becaplermin; D001120:Arginine; C031942:argatroban; C032855:nafamostat