Activated protein C reduces the ischemia/reperfusion-induced spinal cord injury in rats by inhibiting neutrophil activation.
OBJECTIVE
To examine whether activated protein C (APC) reduces spinal cord injury in rats by inhibiting neutrophil activation after the transient ischemia.


BACKGROUND
Ischemic spinal cord injury is an important pathologic mechanism leading to the paraplegia observed after surgery to repair aortic aneurysms. Activated neutrophils play a pivotal role in the development of ischemia/reperfusion-induced tissue injury. Recently, the authors have reported that APC, a physiologic anticoagulant, prevents lipopolysaccharide-induced pulmonary vascular injury by inhibiting neutrophil activation. These observations strongly suggest that APC reduces ischemia/reperfusion-induced spinal cord injury by inhibiting neutrophil activation.


METHODS
In rats, spinal cord ischemia was induced by using a balloon catheter placed into the aorta. After the transient ischemia, survival and motor function were evaluated, and histologic examination of the spinal cord was performed by using both hematoxylin-and-eosin staining and 2,3,5, -triphenyltetrazolium chloride (TTC) staining 24 hours after the ischemia. Tissue levels of myeloperoxidase and cytokines, including tumor necrosis factor-alpha (TNF-alpha) and rat interleukin-8, were measured in six experimental groups: sham-operated, control, APC (100 microg/kg, intravenous), dansyl glutamyl-glycyl-arginyl chloromethyl ketone-treated activated factor X (DEGR-F.Xa), a selective inhibitor of thrombin generation (1 mg/kg, intravenous), nitrogen mustard-induced leukocytopenia, and diisopropyl fluorophosphate-treated APC (DIP-APC), active site-blocked APC (100 microg/kg, intravenous). APC, DEGR-F.Xa, and DIP-APC were administered intravenously 30 minutes before aortic occlusion. Control and leukocytopenic rats received saline instead of other drugs.


RESULTS
Pretreatment with APC significantly reduced motor disturbances compared with those in control animals. In contrast, neither DEGR-F.Xa nor DIP-APC had any effect. Microinfarctions, evidenced by the absence of TTC staining and histologic change, were markedly reduced in animals given APC. The increases in the tissue levels of TNF-alpha, rat interleukin-8, and myeloperoxidase in the ischemic part of the spinal cord were significantly reduced in animals that received APC. These levels were not reduced in rats given DEGR-F.Xa or DIP-APC. Leukocytopenia produced effects similar to those of APC.


CONCLUSIONS
APC reduced the ischemia/reperfusion-induced spinal cord injury by inhibiting neutrophil activation. The therapeutic mechanisms of APC might depend on its inhibitory effect on the production of TNF-alpha, which is a potent activator of neutrophils. Although the anticoagulant effects of APC might not be related to its ability to inhibit TNF-alpha production, its serine protease activity appears to be essential in the therapeutic mechanism. APC appears to have potential as a therapeutic agent for prevention of spinal cord injury in patients undergoing aortic aneurysm repair.
Hirose
K
K
First Department of Surgery and the Department of Laboratory Medicine, Kumamoto University School of Medicine, Kumamoto, Japan.
Okajima
K
K
Taoka
Y
Y
Uchiba
M
M
Tagami
H
H
Nakano
K
K
Utoh
J
J
Okabe
H
H
Kitamura
N
N
D000818:Animals; D007238:Infarction / Q000188:drug therapy; D008297:Male; D018375:Neutrophil Activation / Q000187:drug effects*; D011486:Protein C / Q000627:therapeutic use*; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000401:mortality / Q000517:prevention & control*; D013116:Spinal Cord / Q000098:blood supply / Q000187:drug effects / Q000473:pathology; D013119:Spinal Cord Injuries / Q000150:complications / Q000401:mortality / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D015996:Survival Rate
D011486:Protein C