線維素溶解現象の研究

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Fibrinolytic phenomenon is attributed to an increase of plasmin which is activated by the system of proactivator and activator. The fibrin plate method, which is widely used at present for the purpose of measuring plasmin, has some disadvantages when quantitative measurement of a very small amount of plasmin is required. In this report, the fibrin tube method which was newly devised for the purpose of titrating a very small amount of plasmin is described. Moreover, plasma plasmin in normal healthy persons and that of patients with various diseases was measured by this new method and a relation between diseases and their plasmin value is discussed. Thus, in order to appreciate the property of serum and tissue activators, we tried to separate and extract activators from human serum and tissues. After the measurement method of the tissue activators was examined, the value of activator was determined concerning cancer tissues and mucosae of gastric ulcer obtained from the surgical specimens, and gastric mucosae and various organs from sudden death cadavers. Finally, the corresponding relationship between the activator value and the plasmin value is discussed. I. Fibrin-Tube Method. (1) The fibrin obtained from various animals was examined first. The coagulating conditions of the fibrin incubated for 18 hrs. at 37℃. was investigated, and it was found that bovine fibrin was most available in the bovine, canine, pig's and rabbit's fibrin. (2) Astrup's fibrin plate method was then examined. Conditions of fibrinolysis of the plate colored with carmin suspension were investigated, and four types of fibrinolysis could be differentiated in consequence. 1) Type I : Saucer form- the lysis which does not reach to the base of the petri dish. 2) Type II : Crater form- the lysis reaches to the base of the petri dish, but the area of lysis is narrower at the base than at the surface. 3) Type III : Complete form- the lysis reaches to the base of the petri dish, and the area of the base is equal to that of the surface. 4) Type IV : Triangle flask form- the lysis reaches to the base of the petri dish, but the area of lysis is larger at the base than at the surface. These four types of lysis could not be differentiate by the non-colored plat method of Astrup's. At the time of practical measurement of the lysed area, the value varied in how to decide the lysed diameter. In the case of type I, particularly, the quantitative measurement of plasmin became impossible. (3) For the purpose of quantitative measurement of plasmin, especially that of low activity such as in healthy normal persons, the fibrin tube method was devised in this report. A mass of fibrin colored with carmin was prepared in glass tubes which had a constant inner diameter, and the material to be measured was piled on the mass. The lysed distance was then measured after incubation. As a result of this examination under various conditions, the following was adopted in this report : a clottable protein concentration of 400mg/dl ; the volume of material used was 1.0 ml ; and measurement of the lysed distance was done after incubation for 18 hrs. at 37℃. The fibrin tube method could be compared with the fibrin plate method by means of multiplying the certain conistants. (4) Lastly, the heated fibrin tube method was attempted. The water bath for 10 minutes at 55℃. was most convenient, because of less degeneration of the fibrin, and satisfactory inactivation of the plasmin in the fibrin mass. II. Comparison of the Plasma Plasmin Value Between Healthy Normal Persons, and Patients with Various Diseases Using the Fibrin Tube Method. (1) The plasmin value was determined by the fibrin tube method devised for the quantitative measurement of a small amount of plasmin. The materials were prepared as follows : Plasma was separated from human blood which was prevented from coagulating by an addition of 1/10 volume of 10% w/v sodium citrate. Precipitate of the plasma at pH 5.2 was redissolved in the original volume of the pH 7.8 HCl-Sodium diethylbarbiturate buffer solution. (2) The plasmin value of a normal person was estimated every tow hours, and it was suggested that the value of plasmin was alterative periodically in a day. Practically, the value was minimum at the time of arising and maximum between 3-5 p.m. Measurement of the plasmin value in 13 normal healthy males and 13 females at early morning provided an average of 1.05mm for the male, and 0.96mm for the female. Maximum value was 1.90mm for both sexes. (3) Estimation of the plasmin value in patients with various diseases at early morning was carried out, on the assumption that the maximum limit in the normal persons was 2.0 mm. One hundred and tow patients, all preoperative, were studied at Chiba university hospital. The diseases involved were gastric cancer, esophagocardiac cancer, gastroduodenal ulcer, and others. The average in plasmin values of nearly all the patients was 1.75mm, with σ at 0.89mm. In the cases of gastric cancer, gastroduodenal ulcer, and esophagocardiac cancer, which there were many, the plasmin value indicated higher than when in a normal healthy state. Liver function of the patients was examined due to an increasing of plasmin being attributed to liver diseases by some authors, but a relation between the plasmin value and the liver function could not be correlated. III. Estimation of the Tissue Activators. Measurement of the tissue activator was carried out as follows : the tissues added to 2 M KSCN were homogenated completely and extracted by shaking for 2hrs. The centrifuged supernatant of the extract was then dialysed thoroughly against pH 7.0, 0.1M phosphate buffer solution over night at 4℃., and the dialysed precipitate was collected by centrifugation. The precipitate was added to the plasminogen solution prepared from human serum and Incubated for 5hrs. at 37℃. Then the mixture was centrifuged and the supernatant was titrated by the fibrin plate method. (1) The tissue activators of various organs from cadavers were estimated with the result indicating no direct relationship between the tissue activator value and the course of death. (2) Intensity of the tissue activators ranged almost as follows : the kidney, the suprarenal gland, the liver, the gastric mucosa, the brain, etc. (3) Tissue activator values in patients with gastric cancer and gastric ulcer were higher than those of the gastric mueosa of the cadavers. (4) A direct relationship could not be found between the pre-operative plasma plasmin value of the patients and the tissue activator value of their surgical specimens. However, the group indicated the high value plasma plasmin had as well as the high tissue activator on an average.
千葉大学の論文
1965-05-28

線維素溶解現象の研究

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