SCIE
SCOPUS
KCI등재
Radio Immuno Assay of Insulin affected by Gelatin in Vivo and Vitro System
저자
김윤수 (Department of Biochemistry, Yonsei University College of Medicine) ; 송정석 (Department of Biochemistry, Yonsei University College of Medicine) ; 전의철 (Department of Biochemistry, Yonsei University College of Medicine)
발행기관
생화학분자생물학회(Korean Society for Biochemistry and Molecular Biology)
학술지명
권호사항
발행연도
1971
작성언어
English
KDC
472
등재정보
SCIE,SCOPUS,KCI등재
자료형태
학술저널
발행기관 URL
수록면
10-12(3쪽)
제공처
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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