Lactadherin and procoagulant activities of red blood cells in cyclosporine induced thrombosis  

作　　者：ZHENG Yi-ning YU Hong-juan HOU Jin-xiao LU Cheng-fang ZHOU Jin 

机构地区：[1]Department of Haematology, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, China

出　　处：《Chinese Medical Journal》2009年第14期1674-1680,共7页中华医学杂志（英文版）

摘　　要：Background The side effects of cyclosporine therapy include thromboembolic complications. However, the mechanisms underlying the hypercoagulable state induced by cyclosporine are not fully understood. Cyclosporine binds to red blood cells （RBCs） with a high affinity in circulation and alters the membranes of RBCs. Therefore, we propose that such alterations in RBCs membranes play a role in cyclosporine-induced coagulopathy and this disorder may be rectified by lactadherin, a phosphatidylserine binding protein. Methods RBCs from healthy adults were treated with various concentrations of cyclosporine. Procoagulant activity of the RBC membrane was measured by the single stage recalcification time and confirmed by detection of tenase and thrombin assembly through enzymatic assays. Inhibition assays of coagulation were carried out in the presence of lactadherin, annexin V or antitissue factor. Phosphatidylserine exposure was detected by flow cytometry and confocal microscopy through binding with fluorescein isothiocyanate （FITC）-Iabeled lactadherin as well as FITC annexin V. Results RBCs treated with cyclosporine demonstrated increased procoagulant activity. Cyclosporine treatment markedly shortened the clotting time of RBCs （（305±10） seconds vs （366±15） seconds） and increased the generation of intrinsic factor Xase （（7.68±0.99） nmol/L vs （2.86±0.11） nmol/L） and thrombin （（15.83±1.37） nmol/L vs （4.88±0.13） nmol/L）. Flow cytometry and confocal microscopy indicated that cyclosporine treatment induced an increased expression of phosphatidylserine on the RBC membrane. Lactadherin was more sensitive in detecting phosphatidylserine exposure of the RBC membrane than annexin V. The modulating effect of procoagulant activity was concomitant with and dependent on phosphatidylserine exposure. Blocking of phosphatidylserine with lactadherin effectively inhibited over 90% of FXa generation and prothrombinase activity and prolonged coagulation time. Conclusions Procoagulant propertiBackground The side effects of cyclosporine therapy include thromboembolic complications. However, the mechanisms underlying the hypercoagulable state induced by cyclosporine are not fully understood. Cyclosporine binds to red blood cells （RBCs） with a high affinity in circulation and alters the membranes of RBCs. Therefore, we propose that such alterations in RBCs membranes play a role in cyclosporine-induced coagulopathy and this disorder may be rectified by lactadherin, a phosphatidylserine binding protein. Methods RBCs from healthy adults were treated with various concentrations of cyclosporine. Procoagulant activity of the RBC membrane was measured by the single stage recalcification time and confirmed by detection of tenase and thrombin assembly through enzymatic assays. Inhibition assays of coagulation were carried out in the presence of lactadherin, annexin V or antitissue factor. Phosphatidylserine exposure was detected by flow cytometry and confocal microscopy through binding with fluorescein isothiocyanate （FITC）-Iabeled lactadherin as well as FITC annexin V. Results RBCs treated with cyclosporine demonstrated increased procoagulant activity. Cyclosporine treatment markedly shortened the clotting time of RBCs （（305±10） seconds vs （366±15） seconds） and increased the generation of intrinsic factor Xase （（7.68±0.99） nmol/L vs （2.86±0.11） nmol/L） and thrombin （（15.83±1.37） nmol/L vs （4.88±0.13） nmol/L）. Flow cytometry and confocal microscopy indicated that cyclosporine treatment induced an increased expression of phosphatidylserine on the RBC membrane. Lactadherin was more sensitive in detecting phosphatidylserine exposure of the RBC membrane than annexin V. The modulating effect of procoagulant activity was concomitant with and dependent on phosphatidylserine exposure. Blocking of phosphatidylserine with lactadherin effectively inhibited over 90% of FXa generation and prothrombinase activity and prolonged coagulation time. Conclusions Procoagulant properti

关 键 词：CYCLOSPORINE red blood cell procoagulant activity LACTADHERIN 

分 类 号：R686[医药卫生—骨科学]