Leukocyte-platelet aggregates in rat peripheral blood after ischemic stroke and reperfusion.
Ischemic stroke and reperfusion (ISR) is associated with an inflammatory response characterized, in part, by the formation of leukocyte-platelet aggregates (LPA). Aggregate formation may amplify the immunologic and hemostatic functions of both cell types and thus exacerbate reperfusion injury after ischemic stroke. LPA formation in peripheral blood may also serve as a biomarker of the severity of injury. However, it is not fully known whether ISR causes LPA formation that can be detected in the peripheral blood. Therefore, the purpose of this study was to measure LPA in the peripheral blood after ISR using a rat model. The filament method was used to perform ISR. Blood was collected from the jugular vein before ischemia, after 4 hours of ischemia, and after 1 hour of reperfusion. Flow cytometry was used to quantify LPA in peripheral blood. Separate ISR groups were treated with tirofiban, a platelet GPIIb/IIIa inhibitor, and fucoidan, a selectin adhesion molecule inhibitor, and analyzed for LPA. Leukocyte CD11b expression and reactive oxygen species production were also analyzed to note the role of polymorphonuclear neutrophilic (PMN) activation on LPA formation. After ISR, LPA levels in peripheral blood were twice as large as preischemic levels. Both GPIIb/IIIa and selectin adhesion molecule inhibition (p < .05) decreased LPA to preischemic values. PMN CD11b expression was increased above baseline but did not differ between groups. Reactive oxygen species production did not differ between groups during reperfusion. These data suggest that ischemic stroke and reperfusion results in an increase in LPA that can be consistently measured in peripheral blood. LPA formation may be a useful biomarker and potential therapeutic target after ischemic stroke and reperfusion.
Ritter
Leslie S
LS
University of Arizona, College of Nursing, Department of Neurology, Sarver Heart Center, PO Box 245023, Tucson, AZ 85724-5023, USA. lritter@nursing.arizona.edu
Stempel
Karl M
KM
Coull
Bruce M
BM
McDonagh
Paul F
PF
D000704:Analysis of Variance; D000818:Animals; D000925:Anticoagulants / Q000627:therapeutic use; D015415:Biomarkers / Q000097:blood; D001792:Blood Platelets / Q000276:immunology*; D002545:Brain Ischemia / Q000150:complications*; D004195:Disease Models, Animal; D005434:Flow Cytometry; D007249:Inflammation; D007962:Leukocytes / Q000276:immunology*; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000097:blood* / Q000209:etiology / Q000276:immunology / Q000517:prevention & control; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology; D010974:Platelet Aggregation / Q000276:immunology; D010975:Platelet Aggregation Inhibitors / Q000627:therapeutic use; D011134:Polysaccharides / Q000627:therapeutic use; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000276:immunology; D012720:Severity of Illness Index; D020521:Stroke / Q000150:complications*; D013997:Time Factors; D000077466:Tirofiban; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000627:therapeutic use
D000925:Anticoagulants; D015415:Biomarkers; D010975:Platelet Aggregation Inhibitors; D011134:Polysaccharides; D017382:Reactive Oxygen Species; D014443:Tyrosine; C007789:fucoidan; D000077466:Tirofiban