線維素溶解酵素系の基礎的ならびに臨床的研究 : 第1報 Plasmin測定法の検討 (Fibrin平板法について)

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As to the fibrin plate method, the effect of fibrin membrane preparation and some experimental conditions for determination on the width of fibrinolytic area was examined. 1. The preparation of fibrin membrane 1) In relation between the amount of fibrinogen in the fibrin membrane and the width of the fibrinolytic area, the latter was almost constant against the fibrinogen concentration within the range of 300-500 mg/dl. But it was significantly expanded under the fibrinogen concentration of 100 mg/dl. No significant difference could be found against the thrombin concentration within the range of 10-100 u./ml. But over 500 u./ml, the width of fibrinolytic area was slightly reduced. There could be found a inversely proportional relation between the thickness of the fibrin membrane and the width of fibrinolytic area, and furthermore, larger standard deviation of estimated value could be found corresponding to the development of the thickness. In relation between the ionic strength of the buffer solution pH 7.3 as solvent and the width of fibrinolytic area, the results with μ= 0.15 buffer was somewhat larger (5-21%) than that with μ= 0.50. 2) Next, the effect of heat treatment, which required to denaturate the plasminogen in the fibrin itself, and the width of fibrinolytic area was examined. It was clarified that: i) the width of fibrinolytic area in the plate treated at 85℃ for 10 minutes was reduced to about 47.3% of that of the standard plate without any heating. ii) the results in the plate with 30 minutes heating at the same temperature was 42.9% of the latter, iii) the value with 50 minutes heating was 35.8%, respectively. Complete denaturation of plasminogen in the plate with heating at 85℃ for 30 minutes could be found. 2. Experimental conditions The effect of the concentration of streptokinase on the fibrinolytic activity of the plasma was examined. The width of fibrinolytic area was slightly increased corresponding to the increase of streptokinase concentration within the range of 100-10,000 u./ml. But some decrease of it could be found at the concentration of streptokinase over 1000,000 u./ml. In relation between the incubation time of the plateat 37℃ and the width of fibrinolytic area, the maximal expanding rate of the fibrinolytic area in every one hour was found at 16 hours incubation (21 mm^2/hr.), and it became a rather constant value (6-6.5 mm^2/hr.) after 20-24 hours of incubation. Next, the relation between the temperature of incubation and the width of fibrinolytic area at 18 hours incubation was examined. At 34℃ incubation, it was about 70.8% of that of 37℃ incubation, while the value of 146% was obtained at 41℃. About 10% deviation was found corresponding to the 1℃ temperature alteration arround 37℃. 3. Reproducibility The standard deviation of the results obtained from the standard plate as the width of fibrinolytic area was±6.1-12.9 mm^2 within the range of 198-248 mm^2, and it was rather diversed corresponding to the expansion of the width. In the heat-treated plate, the value of standard deviation was±14.4-15.8 mm^2 within the range of 278-367 mm^2 of the width, and the more lower reproducibility was confirmed on the heat treated plate. With the results obtained above, the more available fibrin plate is: 100-300 mg/dl of fibrinogen concentration, 10-100 u./ml of thrombin concentration, pH 7.3 buffer solution (μ= 0.15), and 1.3 mm in thickness of fibrin membrane, 30 minutes heating at 85℃. And the more reasonable procedure is: 18 hours incubation at 37℃ (one point check between 18-24 hours), and 100-10,000 u./ml of streptokinase concentration.
社団法人日本泌尿器科学会の論文

線維素溶解酵素系の基礎的ならびに臨床的研究 : 第1報 Plasmin測定法の検討 (Fibrin平板法について)

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