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急性 饑餓마우스의 肝蛋白質, 核酸 및 Guanine Deaminase 活性에 關한 硏究

A Study on The Content of Liver Protein, Nucleic Acids, and Guanine Deaminase Activity of Mouse During Acute Starvation

한국영양학회지 1968년 1권 2호 p.107 ~ 115
朴勝熙, 金昇元,
소속 상세정보
朴勝熙 (  ) - 首都女子師範大學 大學院 家政學科
金昇元 (  ) - 서울대학교 醫科大學 生化學敎室

Abstract


Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either. undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo.
It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacrificed 24,´48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g.
The animals were sacrificed by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin. by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionation of the homogenate (10% `V/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4).
For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10 % trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue.being delipidated by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis.
The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utiliz-
ing the Caraway´s colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser´s method, which was followed by Marmur´s method of purification for DNA and by Chargaff´s method of purification for RNA. The determinations of both DNA and RNA were carried out´by´the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work.
1. It was observed that the body as -well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein.
2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded.
3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the de-cline as compared to the former.
4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue.
5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein.
6. The possible mechanisms involved during in this acute starvation as to bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

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