Increased anticoagulant activity of recombinant thrombomodulin modified with glycosaminoglycan
| Autor: | Masao Ohkuchi, Tomoyuki Koshi, Toshiyuki Edano, Hiroko Tabunoki, Akira Murakami, Susumu Yamamoto, Hironobu Sawada, Hideo Yoshizaki, Yasushi Wada, Keisuke Inoue, Noriko Komine |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Male
viruses Thrombomodulin Pharmaceutical Science Thrombin time Biology Pharmacology Fibrinogen Polymerase Chain Reaction Thromboplastin Thrombin Prothrombinase Thrombin receptor medicine Animals Humans Enzyme Inhibitors Rats Wistar Blood Coagulation Glycosaminoglycans medicine.diagnostic_test Antithrombin Anticoagulants General Medicine DNA Angiotensin III Disseminated Intravascular Coagulation Recombinant Proteins Blood Cell Count Rats Kinetics Biochemistry Rabbits Protein C medicine.drug |
| Zdroj: | Biologicalpharmaceutical bulletin. 21(4) |
| ISSN: | 0918-6158 |
| Popis: | Thrombomodulin (TM) is a thrombin receptor on the endothelial cell surface, effective as an anticoagulant by changing procoagulant thrombin to an anticoagulant one. As rabbit TM with glycosaminoglycan (GAG) has a more potent anticoagulant activity than that without GAG, we expressed recombinant GAG-modified urinary thrombomodulin (GAG-UTM) in C-127 cells. The effect of an additional GAG chain on anticoagulant activity was investigated in comparison with unmodified recombinant UTM (r-UTM). In vitro, the activity of cleavage of fibrinogen by thrombin or prothrombinase activity was more potently depressed by GAG-UTM than by r-UTM, and the generation of activated protein C by TM-thrombin complex was accelerated by GAG modification. The acceleration of antithrombin III-dependent anticoagulant activity was shown only by GAG-UTM. Parameters like thrombin time, prothrombin time and activated partial thromboplastin time in human plasma were prolonged by GAG-UTM more than by r-UTM. In vivo, the effect of GAG-UTM and r-UTM in endotoxin-induced disseminated intravascular coagulation (DIC) rats was investigated using hematological parameters. GAG-UTM and r-UTM significantly reduced the decrease in fibrinogen and platelet number induced by endotoxin at the dosage of 0.1 and 1.0 mg/kg/h, respectively, suggesting that the antithrombotic effect of GAG-UTM in endotoxin-induced DIC rats was 10-fold as potent as that of r-UTM. GAG-UTM reduced the prolongation of the bleeding time induced by endotoxin, while r-UTM accelerated it. These results suggest that the addition of a GAG chain may increase availability as an anticoagulant. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|