| Co-Investigator(Kenkyū-buntansha) |
TAKADA A. Hamamatsu Univ., Sch. Med., Department of Physiology, Professor, 医学部, 教授 (80092980)
URANO T. Hamamatsu Univ., Sch. Med., Department of Physiology, Assistant, 医学部, 助手 (50193967)
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| Research Abstract |
We have developed an enzyme immunoassay for the measurements of fibrinolytic parameters such as tissue plasminogen activator (t-PA) and urokinase (UK). A silicone piece coupled with first antibody was incubated with a sample. After incubation the silicone piece was washed and then incubated with a solution containing beta-D-galactosidase labeled second antibody. The activity of beta-D-galactosidase was measured. Using a monoclonal antibody against PAI-1 as a first antibody and beta-D-galactosidase labeled anti- t-PA Fab' fragments as a second antibody, we developed a new assay system to measure the concentrations of total PAI-1, free PAI-1 and t-PA-PAI-1 complex. The amounts of t-PA-PAI-1 complex were measured from the plasma, and those of total PAI-1 were measured from plasma preincubated with t-PA, and those of free PAI-1 were obtained by subtractions of the amounts of t-PA-PAI-1 complex from those of total PAI-1. Plasma levels of t-PA and t-PA-PAI-1 complex increased with increase in age. Plasma levels of t-PA were higher in males than in females, and those in females gradually increased with increase in age, reaching the same levels as those of males at their 60s. Plasma levels of t-PA-PAI-1 complex and free PAI-1 were also higher in males than in females. Then circadian rhythm (8:30, 10:30, 12:30, 14:30 and 16:30 h) of fibrinolysis was measured. Euglobulin lysis time (ELT) showed longest time at 8:30 to 10:30 h and shortest time at 16:30 h. ELT was always shorter in females than in males, The highest level of t-PA or free PAI-1 was obtained in the morning and the lowest was in the evening. These results indicate that the fibrinolytic parameters were balanced at higher levels in plasma in males than in females, but the fibrinolytic activity was higher in females than in males.
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