Elevated fibrinogen fragment levels in uremic plasma inhibit platelet function and expression of glycoprotein IIb-IIIa.
Hemostatic dysfunction is frequently noted in uremia, but the mechanisms responsible for it are poorly understood and are assumed to be multifactorial. Preliminary findings from our laboratory suggest that elevated levels of circulating fibrinogen fragments (FF) might contribute to the hemostatic defect in uremic patients. Defibrinated plasma obtained from chronic hemodialysis (HD) patients as well as normal subjects were examined by SDS-PAGE and immunoblotting and quantified by an immunoassay. In addition, endogenous FF isolated from normal and uremic plasma using affinity chromatography were examined by flow cytometry for their effect on glycoprotein (GP) IIb-IIIa receptor expression and tested for their ability to inhibit platelet aggregation. The mean FF concentration in uremic plasma (1.14 +/- 0.85 mg/ml) was noted to be eight times greater than in normal plasma (0.15 +/- 0.01 mg/ml) (P < 0.05). Moreover, the mean FF level decreased by 48.25% following HD (from 1.14 +/- 0.85 mg/ml to 0.59 +/- 0.33 mg/ml; P < 0.05). SDS-PAGE and immunoblotting experiments showed that the decrease was observed in both medium-sized (20-60 kDa) as well as large (>100 kDa) FF. Further, FF isolated from uremic plasma inhibited platelet aggregation by (46.8 +/- 18.1)% (P < 0.05) and the GP IIb-IIIa receptor expression by (28.0 +/- 7.6)% (P < 0.05 vs. control). The results show that (1) FF levels are elevated in uremic plasma, (2) HD results in significant decrease in FF and (3) endogenous FF inhibit platelet function, presumably via competitive binding to the fibrinogen receptor GP IIb-IIIa. The decrease in plasma levels of FF > 100 kDa following HD suggests that adsorption to the dialysis membrane contributes to their removal.
Thekkedath
Usha R
UR
Research and Medical Service, George E. Wahlen Department of Veterans Afairs Medical Center, and University of Utah School of Medicine; Salt Lake City, UT 84148, USA.
Chirananthavat
Thanit
T
Leypoldt
John K
JK
Cheung
Alfred K
AK
Mohammad
Syed F
SF
D000368:Aged; D000369:Aged, 80 and over; D001667:Binding, Competitive; D001792:Blood Platelets / Q000378:metabolism* / Q000473:pathology; D002846:Chromatography, Affinity / Q000379:methods; D002908:Chronic Disease; D005340:Fibrinogen / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008875:Middle Aged; D010974:Platelet Aggregation; D010979:Platelet Function Tests; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000096:biosynthesis*; D011485:Protein Binding; D006435:Renal Dialysis / Q000379:methods; D014511:Uremia / Q000097:blood* / Q000473:pathology / Q000628:therapy
D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen