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167 The relationship between cytokine concentrations and hemostatic abnormalities in liver cirrhosis
50
POSTER PRESENTATIONS
P-VII F i b r i n o l y s i s i n N o n - V a s c u l a r D i s e a s e s
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PLASMA AND PERITONEAL FIBRINOLYSIS DURING ABDOMINAL SURGERY 1 lvarsson M-L, 1 H o l m d a h l L, 1 E r i k s s o n E, 2 Risberg B lDept, of surgery, 0stra Hospital, university of GOteborg, GOteborg, and 2Dept. of surgery, Malm0 University Hospital, University of Lend, Malta0, Sweden.
FIBRINOLYSlS, THROMBIN GENERATION AND ENDOTHELIAL CELL DAMAGE IN PATIENTS WITH ANTIPHOSPHOLIPID ANTIBODIES. Reverter JC, Monteagudo J, Tbssies D, "Font____J_J,Mara.qall S, Escolar G_, "lngelmo M, Ordinas A. Dept. of Hemotherapy and Hemostasis, and "Autoimmune Diseases Unit. Hospital Clinic. Barcelona. Spain
Peritoneal fibrinolysis is important in the early formation of post surgical adhesions. The aim of the present study was to determine plasma and legal fibrinolytic capacity during surgery in inflamed and normal peritoneum. Plasma samples (from 39 patients of which 7 had peritonitis) and peritoneal biopsies (from 12 patients of which 7 had peritonitis) were taken during surgery when the abdomen was opened, and when the abdomen was closed. The fibrinolytic parameters tissue-type plasminogen activator antigen (t-PA ag) and activity (t-PA act), plasminogen activator inhibitor (PAl-l) was analysed in plasma, t-PA act was analysed in the peritoneal biopsy. The fibrinolytic components were analysed with ELISA technics. Data are given as median, (inter quartile range). In non-peritonitic patients there was a significant increase in the plasma levels of PAId ag 8,6 (6,6) at the beginning versus 22,9 (34,7) at the end of operation. There was also a significant increase of t-PA ag 7,9 (3,2) vs. 8,6 (3,1) and a significant decrease of t-PA act 22,7 (10,3) vs.7.7 (32,5), during surgery. In the peritoneal biopsies the t-PA act was significant reduced 82,3 vs.33,8 during surgery. In patients with peritonitis compared to those without there was a significant higher plasma level of both t-PA ag 15,5 (2,0) vs. 7,9 (3,2) and PAI-1 ag 22,4 (17,9) vs. 8,3 (6,6) at the start of operation. In the peritoneal biopsy there was a significant lower level of t-PA act. at the beginning 38,5 vs.82,3 and at the end 11,5 vs. 33,8 of operation in peritonitic patients. In conclusion there was an decrease in systemic and local fibrinolysis during surgery, as a respons to trauma. In patients with peritonitis the local fibrinolytic activity seemed to be reduced but the systemic fibrinolytic activity was unchanged. The legal inhibition of the fibrinolysios may be important in adhesion formations.
Several mechanisms including abnormalities in the fibrinolytic system have been proposed to explain the association between antiphospholipid antibodies (aPL) and thrombosis. To investigate fibrinolysis components in aPL patients and their relationship with clinical data, thrombin generation and endothelial cell damage markers we studied: 1) 46 patients with systemic lupus erythematosus (SLE) (16 with aPL and previous thrombosis, 12 with aPL but not thrombosis, and 18 without aPL), 2) 14 patients with primary antiphospholipid syndrome (PAS), 3) 6 asymptomatic subjects with aPL, and 4) 45 controls. Blood samples were obtained in the morning without venocclusion. Tissue-type plasminogen activator (tPA), type 1 plasminogen activator inhibitor (PAl-l), plasminogen and a2-antiplasmin were measured by chromogenic assays. Prothrombin fragment 1 + 2 (F1,2), thrombin-antithrombin III complexes (TAT) and thrombomodulin (TM) were determined by ELISA. No differences were seen among the groups in plasminogen, a2-antiplasmin and tPA levels. Increased PAl-1 was observed in plasmas from patients with SLE (p < 0.01) or with PAS (p < 0.01). In SLE patients, there was no correlation between PAl-1 and the presence of aPL or thrombosis. TM was elevated in 39.1% SLE and in 35.7% PAS patients. Patients with elevated TM had significantly higher PAl-1 levels (p < 0.01). F1,2 and TAT levels were elevated in patients with SLE, aPL and thrombosis or with PAS (p<0.01 in both). No relationship was found between PAl-1 and F~+2 or TAT. These results indicate that fibrinolysis seems not related with thrombotic risk in aPL patients and PAl-1 levels are related with endothelial damage markers.
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168
TIIE RELATIONSHIP BETWEEN CYTOKINE CONCENTRATIONS AND HEMOSTATIC ABNORMALITIES IN LIVER CIRRHOSIS
Kvumz Soon Somz. Anna Lee, Q Eun Park..Seur~ Mooj~',e, Oh Hun Kwo~ Department of Clinical Pathology, Yonsei University, College of Medicine, Seoul Korea
Elevated thrombin/antithrombin III(TAT) complex and elevated D-dimer levels have been reported in liver cirrhosis, indicating that cirrhotics have both increased thrombin generation and increased plasmin formation, A number of factors that are elevated in various inflammatory and vascular diseases including the cytoldnes tumor necrosis factor-ct(TNF-a) and interleuldn-l(IL-1), endotexin(LPS), transforming growth factor-13(TGF-I~), and thromhin can stimulate plasminogen activator inhibiter(PAI-1) production by endothelial cells in vitro. Moreover, both in vitro and in vivo investigations have suggested that TNF is an important mediator of the ~ct~vation of coagulation. However, the relationship between cytokines and hemostatic abnormalities in liver cirrhosis is unknown. Plasma concentrations of TNF-, IL-6, TAT and PAI-1 by using enzyme linked immunosorbent assay were determined in 50 patients with cirrhosis(alcoholic=l, postviral= 35, cryptogenic=14) who were at different stages(A=3, B=21, C=26) of Child classification and results were compared to those obtained in 24 healthy subjects. The IL-6 and TNF- levels in patients with cirrhosis were significantly increased compared with those in healthy subjects(median [intarquantile ranges]: 15.96 [8.69-49.79] vs. 0.73 [0.37-1.52] pg/ml, p<0.0001, 5.30 [3.60-10.85] vs. 1.10 [0.77-2.67] pg/ml, p<0.05, respectively). The TAT and PAI-1 levels in patients with cirrhosis were also significantly increased compared with those in healthy subjects(3.95 [3.2-9.15] vs. 2.35 [2.20-2.65] g/l, p<0.001, 32.45 [17.5-49.8] vs. 12.25 [4.7-24.9] ng/ml, p<0.001, respectively). The TNF-0t level was positively correlated with TAT(r=0.5979, p<0.001). The IL-6 level was also positively correlated with those of TNF-a(r=0.3436, p<0.05) and TAT(r=0.3982, p<0.05). However, the PAI-1 level did not show any correlation with cytokines or TAT. We conclude from this study that tumor necrosis factor could play an important part in the activation of hemostatic mechanism in liver cirrhosis, a condition commonly associated with intravascular coagulation.
KETANSERIN CORRECTS SOME PROCOAGULANT CONSEQUENCES OF LONG-TERM rHuEPO TREATMENT 1 1 1 2 Borawski J, Rydzewski A, Mazerska M, Azzadin A, 2Buczko W, IMy~liwiec M INephrology and 2Pharmacodynamics Depts, Medical School, Bialystok, Poland Serotonin (5-hydroxytryptamine; 5-HT) is involved in some changes in haemostasis in initial weeks of recombinant human erythropoietin (rHuEPO) therapy. In the present study, the long-term haemostatic effects of rHuEPO with special relation to 5-HT, were investigated in 15 chronic haemodialysis (HD) patients. Furthermore, w e studied some effects of ketanserin: a blocker of peripheral 5-HT2A receptors, in patients receiving rHuEPO for 8 months. Whole blood platelet aggregation (electric impedance method) was markedly enhanced in response to ristocetin (P<0.005) and collagen (P<0.001) as compared with normals and HD patients not receiving rHuEPO. 5-HT levels in platelets and whole blood that were raised in the 2nd month of rHuEPO (P<0.001), returned to the pretreatment values. A 14-day oral ketanserin administration produced prolongation of the bleeding time (P<0.05), the fall in fibrinogen concentration (P<0.05) and ristocetin-induced platelet aggregation (P<0.005). There were no changes in collagen-, adenosine diphosphate and arachidoniC acidinduced platelet aggregation, or 5-HT levels, euglobulin clot lysis time, or tissue plasminogen activator (tPA) and its inhibitor (PAl-l) concentration. We conclude that long-term rHuEPO patients may be at increased risk of thrombosis. However, the benefit from apparent antithrombotic effects of ketanserin must be carefully weight against a previously reported decrease in serum EPO concentration and inhibition of erythropoiesis in these already anaemic patients.
POSTER PRESENTATIONS
P-VII F i b r i n o l y s i s i n N o n - V a s c u l a r D i s e a s e s
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166
PLASMA AND PERITONEAL FIBRINOLYSIS DURING ABDOMINAL SURGERY 1 lvarsson M-L, 1 H o l m d a h l L, 1 E r i k s s o n E, 2 Risberg B lDept, of surgery, 0stra Hospital, university of GOteborg, GOteborg, and 2Dept. of surgery, Malm0 University Hospital, University of Lend, Malta0, Sweden.
FIBRINOLYSlS, THROMBIN GENERATION AND ENDOTHELIAL CELL DAMAGE IN PATIENTS WITH ANTIPHOSPHOLIPID ANTIBODIES. Reverter JC, Monteagudo J, Tbssies D, "Font____J_J,Mara.qall S, Escolar G_, "lngelmo M, Ordinas A. Dept. of Hemotherapy and Hemostasis, and "Autoimmune Diseases Unit. Hospital Clinic. Barcelona. Spain
Peritoneal fibrinolysis is important in the early formation of post surgical adhesions. The aim of the present study was to determine plasma and legal fibrinolytic capacity during surgery in inflamed and normal peritoneum. Plasma samples (from 39 patients of which 7 had peritonitis) and peritoneal biopsies (from 12 patients of which 7 had peritonitis) were taken during surgery when the abdomen was opened, and when the abdomen was closed. The fibrinolytic parameters tissue-type plasminogen activator antigen (t-PA ag) and activity (t-PA act), plasminogen activator inhibitor (PAl-l) was analysed in plasma, t-PA act was analysed in the peritoneal biopsy. The fibrinolytic components were analysed with ELISA technics. Data are given as median, (inter quartile range). In non-peritonitic patients there was a significant increase in the plasma levels of PAId ag 8,6 (6,6) at the beginning versus 22,9 (34,7) at the end of operation. There was also a significant increase of t-PA ag 7,9 (3,2) vs. 8,6 (3,1) and a significant decrease of t-PA act 22,7 (10,3) vs.7.7 (32,5), during surgery. In the peritoneal biopsies the t-PA act was significant reduced 82,3 vs.33,8 during surgery. In patients with peritonitis compared to those without there was a significant higher plasma level of both t-PA ag 15,5 (2,0) vs. 7,9 (3,2) and PAI-1 ag 22,4 (17,9) vs. 8,3 (6,6) at the start of operation. In the peritoneal biopsy there was a significant lower level of t-PA act. at the beginning 38,5 vs.82,3 and at the end 11,5 vs. 33,8 of operation in peritonitic patients. In conclusion there was an decrease in systemic and local fibrinolysis during surgery, as a respons to trauma. In patients with peritonitis the local fibrinolytic activity seemed to be reduced but the systemic fibrinolytic activity was unchanged. The legal inhibition of the fibrinolysios may be important in adhesion formations.
Several mechanisms including abnormalities in the fibrinolytic system have been proposed to explain the association between antiphospholipid antibodies (aPL) and thrombosis. To investigate fibrinolysis components in aPL patients and their relationship with clinical data, thrombin generation and endothelial cell damage markers we studied: 1) 46 patients with systemic lupus erythematosus (SLE) (16 with aPL and previous thrombosis, 12 with aPL but not thrombosis, and 18 without aPL), 2) 14 patients with primary antiphospholipid syndrome (PAS), 3) 6 asymptomatic subjects with aPL, and 4) 45 controls. Blood samples were obtained in the morning without venocclusion. Tissue-type plasminogen activator (tPA), type 1 plasminogen activator inhibitor (PAl-l), plasminogen and a2-antiplasmin were measured by chromogenic assays. Prothrombin fragment 1 + 2 (F1,2), thrombin-antithrombin III complexes (TAT) and thrombomodulin (TM) were determined by ELISA. No differences were seen among the groups in plasminogen, a2-antiplasmin and tPA levels. Increased PAl-1 was observed in plasmas from patients with SLE (p < 0.01) or with PAS (p < 0.01). In SLE patients, there was no correlation between PAl-1 and the presence of aPL or thrombosis. TM was elevated in 39.1% SLE and in 35.7% PAS patients. Patients with elevated TM had significantly higher PAl-1 levels (p < 0.01). F1,2 and TAT levels were elevated in patients with SLE, aPL and thrombosis or with PAS (p<0.01 in both). No relationship was found between PAl-1 and F~+2 or TAT. These results indicate that fibrinolysis seems not related with thrombotic risk in aPL patients and PAl-1 levels are related with endothelial damage markers.
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168
TIIE RELATIONSHIP BETWEEN CYTOKINE CONCENTRATIONS AND HEMOSTATIC ABNORMALITIES IN LIVER CIRRHOSIS
Kvumz Soon Somz. Anna Lee, Q Eun Park..Seur~ Mooj~',e, Oh Hun Kwo~ Department of Clinical Pathology, Yonsei University, College of Medicine, Seoul Korea
Elevated thrombin/antithrombin III(TAT) complex and elevated D-dimer levels have been reported in liver cirrhosis, indicating that cirrhotics have both increased thrombin generation and increased plasmin formation, A number of factors that are elevated in various inflammatory and vascular diseases including the cytoldnes tumor necrosis factor-ct(TNF-a) and interleuldn-l(IL-1), endotexin(LPS), transforming growth factor-13(TGF-I~), and thromhin can stimulate plasminogen activator inhibiter(PAI-1) production by endothelial cells in vitro. Moreover, both in vitro and in vivo investigations have suggested that TNF is an important mediator of the ~ct~vation of coagulation. However, the relationship between cytokines and hemostatic abnormalities in liver cirrhosis is unknown. Plasma concentrations of TNF-, IL-6, TAT and PAI-1 by using enzyme linked immunosorbent assay were determined in 50 patients with cirrhosis(alcoholic=l, postviral= 35, cryptogenic=14) who were at different stages(A=3, B=21, C=26) of Child classification and results were compared to those obtained in 24 healthy subjects. The IL-6 and TNF- levels in patients with cirrhosis were significantly increased compared with those in healthy subjects(median [intarquantile ranges]: 15.96 [8.69-49.79] vs. 0.73 [0.37-1.52] pg/ml, p<0.0001, 5.30 [3.60-10.85] vs. 1.10 [0.77-2.67] pg/ml, p<0.05, respectively). The TAT and PAI-1 levels in patients with cirrhosis were also significantly increased compared with those in healthy subjects(3.95 [3.2-9.15] vs. 2.35 [2.20-2.65] g/l, p<0.001, 32.45 [17.5-49.8] vs. 12.25 [4.7-24.9] ng/ml, p<0.001, respectively). The TNF-0t level was positively correlated with TAT(r=0.5979, p<0.001). The IL-6 level was also positively correlated with those of TNF-a(r=0.3436, p<0.05) and TAT(r=0.3982, p<0.05). However, the PAI-1 level did not show any correlation with cytokines or TAT. We conclude from this study that tumor necrosis factor could play an important part in the activation of hemostatic mechanism in liver cirrhosis, a condition commonly associated with intravascular coagulation.
KETANSERIN CORRECTS SOME PROCOAGULANT CONSEQUENCES OF LONG-TERM rHuEPO TREATMENT 1 1 1 2 Borawski J, Rydzewski A, Mazerska M, Azzadin A, 2Buczko W, IMy~liwiec M INephrology and 2Pharmacodynamics Depts, Medical School, Bialystok, Poland Serotonin (5-hydroxytryptamine; 5-HT) is involved in some changes in haemostasis in initial weeks of recombinant human erythropoietin (rHuEPO) therapy. In the present study, the long-term haemostatic effects of rHuEPO with special relation to 5-HT, were investigated in 15 chronic haemodialysis (HD) patients. Furthermore, w e studied some effects of ketanserin: a blocker of peripheral 5-HT2A receptors, in patients receiving rHuEPO for 8 months. Whole blood platelet aggregation (electric impedance method) was markedly enhanced in response to ristocetin (P<0.005) and collagen (P<0.001) as compared with normals and HD patients not receiving rHuEPO. 5-HT levels in platelets and whole blood that were raised in the 2nd month of rHuEPO (P<0.001), returned to the pretreatment values. A 14-day oral ketanserin administration produced prolongation of the bleeding time (P<0.05), the fall in fibrinogen concentration (P<0.05) and ristocetin-induced platelet aggregation (P<0.005). There were no changes in collagen-, adenosine diphosphate and arachidoniC acidinduced platelet aggregation, or 5-HT levels, euglobulin clot lysis time, or tissue plasminogen activator (tPA) and its inhibitor (PAl-l) concentration. We conclude that long-term rHuEPO patients may be at increased risk of thrombosis. However, the benefit from apparent antithrombotic effects of ketanserin must be carefully weight against a previously reported decrease in serum EPO concentration and inhibition of erythropoiesis in these already anaemic patients.