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Gelatin이 가토 insulin분비에 미치는 실험적 연구
全毅哲 최신의학사 1971 最新醫學 Vol.14 No.1
At the present time gelatin is used widely in the dietary control of obesity because of its low calorie content. Studies on rabbits in our laboratory have demonstrated that gelatin has an hyperinsulinemic effect on glucose tolerance, and have led us to investigate possible answers to the following questions. (1) What is the component of gelatin which causes the development of the hyperinsulinemic glucose tolerance, and how we can illustrate its mechanism. (2) Are there any quantitative relationships between the glucose and insulin levels in the blood of rabbits receiving gelatin as compared with the controls. In the view of these considerations, the present investigation was undertaken to obtain possible answers to the questions. Five experiments were done using 2 kg rabbits which had been fed bean curd refuse with fresh vegetables, ad libitum. In Experiment 1, four groups of experiments were done as follows: Group 1: After the animals fasted for 12 hours, a glucose tolerance test (Exton-Rose method) was done on each normal animal and on adrenalectnmized animals. Group 2: The glucose tolerance test was done with a simultaneous administration of glucose and gelatin. Group 3: The glucose tolerance test was done 2.5 hours. after the administration of 15 gm of gelatin in 30 ml of water, by stomach tube. The control animals were given 30 ml of water. Group 4: Instead of gelatin, the animals were given an amino acid mixture of gelatin constituent which was equivalent to 15 gm of gelatin. Blood for the measurement of glucose was removed at 0, 30, 60, 90 and 120 minute intervals from a vein of the rabbit's ear. Plasma glucose was determined by the Folin-Wu micro method. Using the method proposed by C. N. Hales and P. J. Randle, a quantitative radioimmuno assay of the plasma insulin was made. Human insulin and 1125 labelled pig insulin, along with insulin binding reagents, were obtained from the Cea-Cen-Sorin Company in France. A standard calibration curve was prepared. In Experiment II, the animals were given an amino acid mixture of gelatin constituent which was administered in group 4 of experiment 1 by adding 50, 100, 150, 200, 400 and 800 mcg of Zn as ZnC12 in 30 ml of water, in contrast to another group of animals which were given a single dose of Zn as ZnCl2 in 30 ml of water. In Experiment III, determination of Zn in gelatin was done using the method proposed by A. S Prasad. In Experiment IV. 200 mcg of Znss as ZnC12 was given in 30 ml of water. Blood, liver and pancreas for the measurement of radioactivity was removed at 0, 30, 60, 90, 120 and 180 minute intervals immediately after decapitation of the rabbit. Radioactivity was determined from the sample by using Well-type-crystal scintillation counter. In order to find out distribution of Znss in the pancreas cell, a radioaudiography was performed on the pancreas, obtained 150 minutes after Znss administration. In Experiment V, digestive enzymes, amylase, lipase, trypsinogen and chymotrypsinogen activities of the pancreas, 150 minutes after Zn administration, were determined, It is evident that, as the glucose level in the blood increased, the secretion of insulin also increased until a plateau was reached 60 minutes after the administration of glucose. From this 60 minute plateau the insulin level returned , to its original value. in 60 minutes. However, the glucose and gelatin were given to the rat at the same time there was no difference observed in the glucose levels in the gelatin treated rats and the control rats. In remarkable contrast, however, rabbits given 15 gm of gelatin, 150 minutes before being given glucose, showed a 30 minute secretion level of insulin which was twice that of the control -animals. This occurred even though the glucose level in the two groups remained the same over the same interval of time. Thus the effect is due directly to the gelatin.. However, the effect is not due to the amino acid constituent of gelatin, but due to Zn which is a constituent of gelatin, because this fact was also true when 200 mcg. of ZnClz were given instead of gelatin. Meanwhile no remarkable differences were observed in the glucose levels between the group ZnCI2 with glucose and the gelatin treated group. Low concentration of Zn (50-100 mcg) does not, seem to be responsible, on. the other hand a high conceptration (400-800 mcg) of Zn inhibits the secretion of insulin. The responsible concentration of Zn being 150-200 mcg for secretion of insulin. The Zn content of gelatin averaged 110 mcg. per gm of gelatin. C. P. M. of Zn65 in plasma, after administration of Zn65 was increased up to 90 minutes, then a saturated phenomena was observed. However, a continuous increase of C. P. M. of Zn was shown in the liver and pancreatic tissue, with a little more rapid rate of uptake in liver than in pancreatic tissue. It. was confirmed that the Zn65 was mainly localized in the acinar cells of the pancreas. The digestive enzyme activity was not affected at all. We had a postulate as to which component of gelatin was responsible for the secretion of insulin, and the results partially confirmed the postulate that glucose, insulin and Zn levels, in cellular process, were related.
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朴東浩,全毅喆,韓景植,김완식,Ian. S. Robb 中央醫學社 1968 中央醫學 Vol.14 No.3
Recently, many investigations have been made regarding the way in which the newborn infant changes over in a first minutes of life from a passive intrauterine existence to more active extrauterine life. Establishment of an extra-uterine circulatory system involves separating the systemic and pulmonary circulation and the obliteration of the placental circulation. When should the cord be clamped in relation to its haemodynamic and respiratory effects on the infant, if the placental blood is transfused into the infant, in the first week of life there is a significant increase in the erythrocyte count and hemoglobin as compared to the infant in which clamping was early. Furthermore, failure of the infant to receive its placental blood may lead to a deficiency of iron during the first year of life, and if the baby does not receive this placental blood a hypovolaemic state may ensure. The principle of possibly a more physiological method is to extract infant and placenta together, without. cutting the cord, and then to establish an environment which allows the placental blood volume to be transfused according to the infant's circulatory demands. This has been done Severance Hospital, Yonsei University Medical College, Department of Anesthesiology. Out of a series of 735 delivered cases from October 1966 to May 1967, it was used in 20 infants delivered by caesarian section and 30 normally delivered infants, after clamping of the umbilical cord from the moment of birth to spontaneous birth of the placenta, and it is very easy to carry out. There seems to be no essential difference between the blood volume transfused after delivery by caesarian section and after normal vaginal delivery. In most of transfused infants this consisted about 23% of total blood volume and was 50 to 100ml. From the view point of the haemoglobin, if the infant is deprived of this placental blood there is a significant reduction of the haemoglobin as compared to infants in which clamping was delayed. The described procedure is still experimental, and it is too early to say whether better results are possible. A larger series has been started and we are studying, in correlation with the obstetricians, the physiology of respiratory and circulatory changes in relation to different forms of delivery.
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Radio Immuno Assay of Insulin affected by Gelatin in Vivo and Vitro System
김윤수,송정석,전의철 생화학분자생물학회 1971 BMB Reports Vol.2 No.2
At the present time gelatin is widely in the dietary control of obesity because of its low caloric content. Recent studies in the pancreas of the rat have demonstrated that gelatin has a biosynthetic effect on trypsin and chymotrypsin. The mechanism of this effect is unknown. Studies in rabbits in our laboratory have demonstrated that gelatin also has an hyperinsulinemir. effect on glucose tolerance, and have led us to investigate possible answers to the following questions. (1) What is the component of gelatin which causes the development of the hyperinsulinemic glucose tolerance ? (2) Are there any quantitative relatioships between the glucose and insulin levels in the blood of rabbits receiving gelatin as compared with the controls ? (3) What is the effect on human beings of the hyperirsulinemic factor found in gelatin ? We postulated that gelatin then may contain a regulatory secretion factor which affects biosynthesis and pancreatic secretion of insulin. To search for this hypersecretory factor in gelatin a quantitative radioimmuno assay of insulin as it was affected by the factors of glucose and ZnCl₂ was made in vitro. Four experiments were done using 2 ㎏ rabbits. which had been fed bean curd refuse with fresh vegetables. In experiments I, after the animals were fasted for 12 hours, a glucose tolerance test (Exton-Rose method) was done on each. In Experiment II, the glucose tolerance test was done 2. 5 hours after the administration of 15 gm of gelatin in 30m1 of water by stomach tube. The control animals were given 30 ㎖ of water. In Experiment III, instead of the gelatin, the animals were given 200 ㎎ of zinc as ZnCl₂ in the 30 ㎖ of water. Blood for the measurement Of glucose was removed at 0, 30, 60, 90, 120 minutes intervals from the rabbit's ear vein. Plasma glucose was determined by the Folin-Wu micro method. Using the method proposed by C. N. Hales and P. J. Randle, a quantitative radioimmunoassay of the plasma insulin was made. ^(-125)I labelled, human insulin and ^(-125)I labelled pig insulin, Tong with insulin binding reagents, were obtained from the CEA-CEN-SORIN comany in France. A standard calibration curve was prepared. In Experiment IV, immediately after decapitation of the rabbit the pancreas was carefully removed and placed in a cold bicarbonate buffer solution (4 ). Obvious lymph nodes were removed from the glandular portion along with adipose tissue. The pancreas then was cut into R-11 ㎎ pieces. 500 ㎎ of pancreas, divides into small pieces, were blotted on filter paper and then placed in 2 ㎖ of the incubation medium. It is evident that, as the glucose level in the blood increased, the secretion of insulin also increased until a plateau was reached 60 minutes after the administration of glucose. From this 60 minutes plateau the insulin level returned to its original value in 60 minutes. However, the glucose level continued to remain high, as much as 170 ㎎ glucose. When glucose and gelatin were given to the rat at the same time there was no difference in the glucbse levels observed in the gelatin treated rats and the control rats. In remarkable contrast, however, rabbits given 15 gm of gelatin 2.5 hours before being given glucose showed a 30 minutes secretion level of insulin which was twice that of the control :animals. This occured even though the glucose level in the two groups remained the same over the 'same interval of time. Thus the effect is due directly to the gelatin. This fact was also true when 200 ㎍ of. ZnCl₂ was given instead of gelatin. Meanwhile no remarkable difference were observed in the glucose levels between the group given ZnCl₂ with glucose at the same time as gelatin and the controls. The maximal excretion sate came at a concentration of 2b ㎍/100㎖. If the ZnCl₂ concentration was increased to more than 26 ㎍/100㎖ concentration, in vitro, there was no corresponding increase in insulin secretion. This would correlate with in vivo experiments in which it was found that the insulin was not secreted at all when 40㎍ of ZnCl₂ was administering by orally to the 2 ㎏ of rabbit, these amount of ZnCl₂ was administered is .equivalent to 33 ㎍ of ZnCl₂ concentration/100 ㎖ of infra and extracellular fluid of rabbit. The study reported here was designated to provide us with information concerning the effect of gelatin and ZnCl₂ on insulin secretion in vivo and in vitro. We had a postulate as to which components of gelatin were involved in the secretion of insulin. The study partially confirmed the postulate that glucose, insulin and Zn levels in blood are related in vivo and in vitro. We are now investigating the relationships. between the glucose, insulin and Zn levels in vitro, at the present time this study is not complete.
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