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再生肝組織의 成長促進性因子에 關한 硏究

Growth Promoting Factor in Regenerated Liver Tissue

최신의학 1965년 8권 1호 p.53 ~ 63
김영홍,
소속 상세정보
김영홍 (  ) - 서울대학교 의과대학 외과학교실 서울대학교 의과대학 암연구소

Abstract


Authors attempted in the present paper to investigate to obtain the growth promoting factor from the regenerated liver tissue of partially hepatectomized rat, and to investigate the effect of it on the growth of cancer and normal tissue of mice.
materials and methods
It is well known that after partial hepatectomy in rats the residual part of the liver regenerates rapidly and reaches the size of a normal liver in 2∼3 weeks. In the present investigation partial hepatectomy was performed by Higgin´s methed. Under ether anesthesia a median incision was made from the xyphoid process 2∼4 ㎝ caudad. Left lateral and median lobes were aseptically removed in toto with only slight loss of blood. This yielded approximately 65∼75% volume of total liver. At 3 days interval up to 15 days postoperatively the rats were sacrificed by ether anesthesia and then removed the regenerated liver tissues.
The cancer strain used was Ehrlich ascites cancer and the tumor transplantation was performed according to Whwang´s methed.
1. The normal and regenerated liver tissues were homogenized adding 4 volumes of the distilled water, centrifuged in a Model PRI International Centrifuge at 15,000 r.p.m., and the precipitate was discarded. To the supernatant 95% ethyl alcohol was added and discarding the alcohol precipitate, the resulting supernatant was evaporated. 5% & 10 % tissue factor solutions for injection were prepared from the residue. And then a series of injections of both the homogenates and tissue factor solutions(5%, 10%) were done once daily. On the 18th day after tumor transplantation, the animals were sacrificed, the tumors being removed and weighed. The growth of the tumors were measured by average length of 2 diameters of the tumors and these were compared with those of control group.
2. Experiment on the growth promoting effect of normal and regenerated liver tissue factor on the cellular mitosis of liver and kidney of normal mice was carried out in the following manner. A single dosis of 0.1 ㎖ of the 10 tissue factor were injected intraperitoneally every day. On every 3 days, animals were sacrificed, the liver and kidney were removed, and fixed in the Canoy´s solution. After dehydration and decalcification, the tissues were embedded in Paraffin, cut in sections of 5 micron thickness, and stained with Hematoxylin-Eosin. Mitosis was counted in 100 fields on each slide with magnified microscope(430×), comparing with those of the control group in which 0.1 ㎖ of the saline solution was injected instead of tissue factor as sham experiment.
Results
As to the growth promoting effect of the homogenate and, tissue factor on the growth of Ehrlich ascites cancer, the preparation obtained from the 3rd-day-regenerated liver shows the- highest effect and the effect of the other preparations decrease as the regeneration progresses.
a. The homogenate has dose-dependent growth promoting effect, 0.1 ㎖ showing the highest and 0.5 ㎖ the lowest accelerating effect.(Table 1.2.3., Fig 1.)
b. The 5% isolated tissue factor has more acceleration effect than the homogenate showing more effective growth with 0.2 ㎖l than 0.1 ㎖.(Table 4. 5., Fig 2.)
c. The 10% isolated tissue factor has more acceleration effect than the 5%, 0.1 ㎖ showing the strongest effect and 0.5 ㎖ the weakest. (Table 6.7.8., Fig 3.)
2. In comparison between promoting effects of normal liver tissue factor and regenerated liver tissue factor on cellular mitoses of liver and kidney of the normal mice, both promote the cellular mitoses of the normal tissues of mice, and the latter being more marked in effect than the former.(Table 9, 10, Fig 4, 5) Further more both factors displayed stronger promotion uppon cellular mitosis of liver than of kidney. The successive daily injection promotes growth gradually up to a peak acceleration and then decreases gradually in the following days.
The tissue specificity involved in this observation is discussed.

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