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KMID : 0378019680110070075
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New Medical Journal
1968 Volume.11 No. 7 p.75 ~ p.88
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The Effect of Injury of Animal Tissues with Carbon Tetrachloride on Tissue Factor Formation
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Abstract
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Since establishing hypothesis that the homeostatic state and orderly growth of normal cells are maintained by the interaction of two forces, one which stimulates growth and the other which retards it, and then if the disturbance occur in the balance between the factors, either the loss of the retarding or the accumulation of the stimulating factor, would lead to uncontrolled growth, by Murphy and Sturm, numerous other investigators have made a study of both growth promoting and growth inhibitory factors extracted from chemical or mechanical damaged tissues as well as various normal tissues.
A series of experiments has been undertaken to investigate the influence of the tissue homogenate and tissue growth promoting and inhibitory factors, extracted from cirrhotic and injured liver induced in rabbit by injecting of carbon tetrachloride on the growth of cancer. And at the same time homogenate and both promoting and inhibitory factors of kidney which associated with cirrhotic and injured liver induced by carbon tetrachloride were also used same investigation.
Materials and Methods
Rabbits, irrespective of age or sex, about 2 kg in weight were obtained, and every three days single dose of 1.32 cc of 25 % carbon tetrachloride in liquid petrolatum per kg of body weight were injected intraperitonealy or subcutaneously after Hoffman and the other´s method.
The liver cirrhosis was noted on 80 to 100 days after first injection of carbon tetrachloride.
The injured liver and kidney were obtained from expired rabbits during treatment with carbon tetrachloride between 2 to 4 weeks.
The cirrhosis and injury of those organs were checked and confirmed histopathologically. Sterile technic was used in the preparation of tissue homogenates and factors as possible. The removed cirrhotic, injured and normal liver and kidney were weighed and macerated with 9 volumes of distilled water in a Waring blender respectively. These homogenate was, then, mashed through a sterile gauze (80 mash. In this method 10 % homogenate was prepared for ready of experiment. Tissue factors were extracted from cirrhotic, injured and normal liver and kidney respectively by Chin´s method. Each livers and kidnies were homogenized adding 4 volumes of the distilled water in Waring blendor. The homogenate was, then, filtered and the filtrate was centrifuged at 15, 000 r.p.m. for 40 minutes in an International centrifuge Model PRI. One volume of 95 % ethyl alcohol was added to one volume of the supernatant and it was left over night in the 4℃of ice box. And again centrifuged for 1, 500 r.p.m. for 20 minutes._ After dry for few days the final super-natant was used as the promoting tissue factor, and the other sediment as the inhibitory factor.
White mice weighing about 20 gm were used throughout the studies. 0.2m1 of Ehrlich ascitis cancer was inoculated sutcutaneously into the right side of the abdominal wall of each mouse for formation of solid form of Ehrlich cancer according to Chin´s method. These, 10/ solution of homogenate, 5 % solution of tissue promoting and inhibitory factors, were started to inject subcutanieously on the left side of abdominal wall, the other side of inoculation of cancer, on the day of the tumor inoculation of the mice and continued once a day for 18 consecutive days.
The daily doses per animal consisted of 0. 1, 0. 2, 0.3 ml each of 10% solution of homogenate and 5 % solution of extracted tissue factors respectively.
The size of the tumor was directly measured every three days on the basis of individual tumor from the skin with calipers and average tumor diameter of each group was calculated from two perpendicular diameters.
On the last day of injection, the 18th day after transplantation of the tumor, animals were sacrificed, the excisional tumor was weighed and the average weight of tumor of each group was calculated. The results of above were compared with the control group which were given a daily of corresponding volume of physiological saline.
Results
1) Effect of the tissue homogenate on the growth of Ehrlich ascitis cancer was generally shown marked promoting action as table 1, 2, 3, 4, 5, 6, Fig. 1 and 2. The 10% homogenate of cirrhotic group of liver and kidney, induced in rabbit by injecting of carbon tetrachloride had the most prominent effect of growth promotion, and was followed by injured group of those. Normal group of those shows the lowest effect.
2) Every group of 5% isolated growth promoting tissue factors accelerated markedly the tumor growth as compared with the controls. It was shown in table 7, 8, 9, 10, 11, 12, Fig. 3 and 4.
The promoting action was most prominent in group of injured liver and kidney tissue factors and was followed by group of normal liver and kidney tissue factors. The group of cirrhotic liver and kidney tissue factor had the lowest effect.
The potentiality of acceleration in every group of this experiment were increased according as increasing of dosage of administration from 0.Icc, 0.2cc to 0.3cc (refer to Table 19)
3) The inhibiting effect of 5% isolated growth inhibitory tissue factors on tumor growth was remarkable in this experiment as well as the promoting effect, which was shown by table 13, 14, 15, 16, 17, 18, Fig. 5 and 6.
But the most forceful inhibiting action was observed in the administration of inhibiting tissue factor which isolated from normal liver and kidney. And which was followed by that of injured liver and kidney, and cirrhotic liver and kidney, but both of latter were almost similar in potentiality
The potentiality of inhibiting factor according to dosage of injection was same as the case of promoting factor. (refer to Table 20)
4) Every effect of liver homogenate, tissue promoting and inhibiting factors extracted from livers were more forceful and roticeable than those of kidney throughout of the studies. {
5) From this experiment it is suggested that in the cirrhotic tissue there is not only a decrease of inhibi-
tory activity but a decrease of promoting factor as well.
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