• 한국식품영양과학회지
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• 제31권5호
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• pp.782-787
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• 2002

본 연구는 한국산 겨우살이 물 추출물과 이것에서 분리한 렉틴 성분을 실험적으로 간암을 유도한 흰쥐에게 투여하여 이들이 간 발암억제 효과 및 그 기전의 일부인 apoptosis와의 관련성을 조사하고자 하였다. 이를 위해 80～90 g 정도로 이유된 .3주령 Sprague-Dawley종을 한국산 겨우살이 물 추출물 투여 (100 $\mu\textrm{g}$/kg BW), 렉틴의 투여 10 ng/kg BW)와 DEN 투여 여부에 따라6군으로 나누어 발암물질 투여 1주일 후부터 주 2회 복강 투여하였다. 총 사육기간은 9주간이었고, 희생시킨 후 간조직을 분리하여 조직 병리학적 변화 측정을 위한 전암성 병변의 지표인 GST-P$^{+}$foci의 수와 면적을 측정하였고, apopotosis와의 관련성을 조사하기 위하여 apoptosis 염색, DNA fragmentation 측정 및 Bcl-2, c-lAP2, caspase-9, caspase .:, fas-L의 발현정도 측정을 위한 western blotting을 실시하였다. 그 결과 다음과 같은 결론을 얻었다. (1) 체중은 발암물질에 의해 식이 섭취가 감소하는 것을 고려하여 제한 섭취를 실시하였으므로 발암물질 투여 에 따른 차이는 없었으며, 겨우살이 또는 렉틴의 투여에 따른 차이도 나타나지 않았다. 간 무게는 겨우살이, 렉틴의 투여에 따른 변화는 보이지 않았으나 발안물질 투여 에 따라 유의적으로 증가하였고, 체중에 대한 상대적인 간 무게에 대한 결과도 간 무게의 결과와 유사하였다. 따라서 본 연구에 사용한 투여 량의 한국산 겨우살이 물 추출물 및 렉틴이 간에 독성을 나타내는 농도가 아닌 것으로 보여진다. (2) GST-P$^{+}$ foci의 수, 면적과 PCNA은 겨우살이 물 추출물과 렉틴의 투여에 따라 모두 유의적으로 감소하였고, 겨우살이 물 추출물과 렉 틴에 따른 차이는 보이지 않았다. (3) 조직화학적으로 apoptosis염색한 결과와 DNA fragmentation을 측정한 결과는 모두 apoptosis에 대한 증거가 나타나지 않았다. 그러나 western blot을 측정한 결과 apoptosis을 억제하는 것으로 알려진 단백질인 Bcl-2와 IAP2는 발암물질 투여에 의해 증가된 발현 정도가 겨우살이 물 추출물이나 렉틴의 투여에 따른 차이를 보이지 않았으며, apoptosis를 촉진하는 단백질인 caspase-9, fas-L의 경우에는 발암물질 투여 에 의 해 증가된 발현 정도가 겨우살이 물 추출물 및 렉틴의 투여시 더욱 증가되었다.

• 한국식품영양과학회지
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• 제30권4호
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• pp.697-702
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• 2001

본 연구는 한국산 겨우살이 물 추출물과 이에서 분리한 렉틴 성분을 실험적으로 간암을 유도한 흰쥐에게 투여하여 이들의 간 발암억제 효과를 조사하고자 하였다. 이를 위해 80~90 g정도로 이유된 3주령 Sprague-Dawley종을 유럽산겨우살이 추출물 투여(10 $\mu\textrm{g}$/kg BW, 100 $\mu\textrm{g}$/kg BW)와 렉틴(1 ng/kg Bw, 10 ng/kg BW)의 투여량 DEN 투여 여부에 따라 12군으로 나누어 발암물질 투여 1주일 후부터 주 2회 복강 투여하고 실험식이를 공급하였다. 총 사육기간은 9주간이었고 희생시킨후 혈청과 간 조직을 분리하여 조직병리학적 변화 측정을 위한 전암성 병변의 지표인 GST-P+ foci의 수와 면적을 측정하였고 생화학적 변화 측정을 위해 간 기능의 지표인 혈청 GOT와 GPT 활성도, 혈청 $\alpha$-L-fucosidase 활성도를 측정하였다. 그결과 다음과 같은 결론을 얻었다. (1)체중은 발암물질에 의해 식이 섭취가 감소하는 것을 고려하여 제한 섭취를 실시하였으나 발암물질 투여나 겨우살이 또는 렉틴의 투여에 따라 약간 감소하는 경향을 보였고 간 무게는 겨우살이 렉틴의 효과는 보이지 않으나 발암물질 투여에 따라 유의적으로 증가하였다. 체중에 대한 상대적인 간 무게에 대한 결과도 간 무게의 결과와 유사하였고 유럽산보다는 한국산 겨우살이 물 추출물의 간 무게와 체중에 대한 간 무게의 비율이 작게 나타나 한국산 겨우살이 물 추출물의 독성이 간에 영향을 작게 미치는 것으로 보여진다 (2) GST-P+ foci의 수나 면적은 겨우살이나 렉틴의 투여에 따라 유의적으로 감소하였고 유럽산과 한국산의 겨우살이 추출물 종류에 따라서는 한국산 겨우살이 추출물을 투여한 군에서 약간 증가하는 경향을 보였으나 한국산 겨우살이 물 추출물의 양을 증가시켜 100 $\mu\textrm{g}$/kg BW 수준으로 투여한 결과 foci의 수나 면적이 유의적으로 감소하여 가장 낮은 값을 보였다. 렉틴의 투여량에 따른 결과도 양이 증가함에 따라 GST-p_ foci의 수와 면적이 감소하였다. (3) 혈청 GOT 활성도는 유의적이지는 않지만 발암물질 투여에 따라 증가하는 경향을 보였고 겨우살이나 렉틴의 투여에 따라유의 으로 증가하였으며 겨우살이와 렉틴에 따라서는 GOT와 비슷한 경향을 나타냈다.을 나타냈다.

• 한국식품위생안전성학회지
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• 제5권4호
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• pp.179-186
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• 1990

이 연구는 수입 축산식품중에 잔류해서 사람에게 해를 줄 수 있는 여러 합성 홀몬의 발암성을 알아보기 위하여 수행한 연구이다. 6주령의 F344 랫드를 5군으로 나누어 발암유발물질인 diethylstilbesterol(DENA)를 체중 kg당 200 mg을 복강 내 투여하고, 시험게시 제2주에서 시험종료시까지 제 1, 2, 3,군에 각각 diethylstilbesterol(DES), ethynylestradio($EE_2$), bovine somatotrophin(BST)을 투여하고, 제 4군에는 발암촉진이 잘 알려진 phenobabital을 투여하였으며, 제 5군은 대조군으로 기초사료만을 급여하였다. 동시에 모든군의 랫드는 D-galactosamine을 체중 kg 당 300 mg을 복강내로 1회 투여하였으며, 간변화의 증폭을 위하여 2/3 간부분 절제를 시험개시 제 5주에 실시하였다. 시험종료시 부검후 랫드의 간장은 GST-P에 대한 면역조직화학적 염색을 실시하여 발암성을 검색하였다. 두개의 합성 에스트로젠 홀몬($EE_2$, DES)은 대조군과 비교하였을 때 발암성이 있는 것으로 생각되어지고, BST는 발암성이 없는 것으로 생각된다.

• Biomolecules & Therapeutics
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• 제3권4호
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• pp.279-287
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• 1995

This study examined the anti-cancer effects of diallyl sulfide(DAS) and/or diallyl disulfide(DDS), major components of garlic oil, with the DEN-PH model in rats, by the numbers and areas per cm$^2$ of induced glutathion S-transferase placental form(GST-P) positive foci and silver-stained nucleolar organizer regions(Ag-NORs) counts per nuclei in liver as indicator. Sprague-Dawley(SD) rats were given the diethylnitrosamine(DEN, 200 mg/kg, i.p.) as initiator and 2 weeks later, in experiment 1, rats were treated with DAS(200 mg/kg, i.g.) and/or DDS(50 mg/kg, i.g.) for 6 weeks, respectively and concomitantly and also were given the same dose of DAS and/or DDS prior to DEN treatment for 2 weeks, and in experiment II, rats were treated with potential cancer promoter, 2-acetylaminofluorene (2-AAF, 20 mg/kg, i.g.). The DAS and/or DDS were treated prior to 2-AAF for 8 weeks, respectively and concomitantly. Then the anti-promoting effects of DAS and/or DDS were assessed. All rats were subjected to the two-thirds partial hepatectomy(PH) at week 3 and sacrificed at week 8. In experiment I, DAS and/or DDS treatment only prior to DEN showed inhibition of the development of GST-P positive foci. In experiment II, DAS and/or DDS treatment prior to 2-AAF promotion showed obvious inhibition of the development of GST-P positive foci in numbers and areas and AgNORs counts. In conclusion, We found DAS and/or DDS had the preventive effects on the hepatocarcinogenesis in rats and the concomitant treatment had some additive effects compared with the each treatment and AgNORs counts correlated well with the preneoplastic hepatic lesion.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2002년도 추계학술대회초록집
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• pp.149-149
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• 2002

Epidemiologically the incidence of liver cancer is markedly sex-differentiated, with a much higher frequency in men than in women. In experimental animals, it is also higher in male than in female irrespective of carcinogen-induced or spontaneous tumors. Therefore, we tried to investigate the modulating effects of sex hormones in experimental hepatocarcinogenesis. For induction of liver tumors, mini-osmotic pump containing diethylnitrosamine at a dose level of 47.5mg was implanted into the peritoneal cavity of the rat at 6 weeks old. To remove the effects of male sex hormones, the animals of group 2 were castrated one week prior to DEN treatment. To see the effects of estrogen, pellet containing 1g or 10g of estradiol-3-benzoate was infused subcutaneously to the animals of group 3 and 4 one week prior to DEN treatment. The pellets were exchanged every 4 weeks until sacrifice. All animals were sacrificed at 26 weeks after DEN treatment. The tumor incidences in group 1 (DEN alone), group 2 (DEN +castration), group 3(DEN +EB 1g) and group 4 (DEN +EB 10g) were 100％ (15/15), 93.3％ (14/15), 85.7％ (12/14) and 66.7％ (10/15), respectively, showing that the value of group 4 is significantly different from that of group 1. Tumor multiplicity data of group 1, 2, 3 and 4 were 5.470.73, 2.800.51, 2.070.41 and 1.670.46, respectively, showing castration or EB treatment reduced number of liver tumors significantly (P<0.001). With immunohistochemistry and Western blotting of ER the expressions were detected in normal adjacent liver cells but decreased or lost in tumor cells. From these results we conclude that female sex hormone, especially estrogen, may act as a liver tumor suppressor, and it seemed that the down regulation of ER may be associated with liver tumor development.

• 대한수의학회지
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• 제40권1호
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• pp.117-129
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• 2000

This study was carried out to examine proliferative activity and expression of c-myc oncoprotein and p2lras in normal and preneoplastic rat livers induced by an in vivo mid-term chemical carcinogenesis assay. Sixty, six-week-old male specific pathogen free Sprague-Dawley male rats were randomly divided into five groups. Group I was received a single intraperitoneal(IP) dose(200mg/kg) of diethylnitrosamine(DEN). Group 2(10 rats) was operated partial hepatectomy(PH) and Group 3 was received IP(200mg/kg) DEN, fed two weeks later with 500ppm of phenobarbital(PB). Group 4 was received IP(200mg/kg) DEN, fed two weeks later 500ppm(PB) and PH at week 3 after the onset of experiment. While group 5(20 rats) was not treated and used as a control group. All the rats were sacrificed at age 14 weeks except 10 rats from group 5 were sacrificed at the onset of experiment. Livers of all rats were examined for 5-bromo-2'-deoxyuridine(BrdU) incoporation, proliferating cell nuclear antigen(PCNA), silver-binding nucleolar organizer regions(AgNORs) counts per nucleus and expression of c-myc oncoprotein and p21ras. Both the number and area of the preneoplastic lesions were significantly(p<0.01) compared to other groups. A significant(p<0.01) increase in immunoreactive cells were detected in preneoplastic hepatocytes in Groups 3 and 4 by PCNA and BrdU immunohistochemical stain. The number of the positive cells were significantly(p<0.05) lower in normal 14-week-old rats than those of 6-week-old rats. The results showed that proliferative activity of the hepatocytes was increased by treatment with DEN, PH and PB. Meanwhile, AgNORs counts per nucleus were significantly(p<0.05) increased in the preneoplastic hepatocytes of rats in both groups 3 and 4. The expression of c-myc oncoprotein and p21ras were more readily localized within the hepatic preneoplastic lesions such as hyperplastic nodules. Especially, group 4 showed significantly (p<0.05) overexpressed levels compared to groups 1 and 3. These findings suggest that PCNA, BrdU and AgNORs are significantly increased and c-myc oncoprotein and p21ras are significantly overexpressed in hepatic preneoplastic lesions induced by mid-term carcinogenesis. So these parameters can be an effective markers for hepatic prencoplastic lesions.

• 대한지역사회영양학회지
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• 제2권5호
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• pp.735-744
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• 1997

The effect of green tea drinking on the hepatocellular chemical cacinogenesis have been studied. Placental glutathione S-transferase(GST-P) positive foci area in a liver tissue, contents of thiobarbituric acid reactive substances(TBARS), total cytochrome P450 and glucose 6-phospphatase(G6P) activity in hepatic microsomes were investigated. Weaning Sprague-Dawley male rats were fed AIN-76A diet with deionized water or green tea infusion, Rats of CTR and CTR+ groups were provided deionized water while GTI and GTI+ groups were provided green tea instead of deionized water for the entire experimental period of 13weeks. Rats of GTP and GTP + groups had deionized water for the first 6 weeks and switched to green tea for the last 7weeks of the experimental period. CTR+, GTI +, and GTP + groups were carcinogen treated groups, Diethylnitrosamine(DEN) was injected as a single dose of 200mg/kg body weight intraperitoneally after 4 weeks of feeding. 2-Acetyla-minofluorene(AAF) was used as a carcinogen proliferater and suppled in the diets of carcinogen treated rats as 0.02% content for the last 6weeks starting from 2weeks after DEN injection. Rats were sacrificed after 13week weeks of feeding. The area and number of GST-P positive foci detected in carcinogen treated rats were decreased by green tea ingestion but when timing and duration of green tea ingestion was delayed after promotion period as in GTP + group, GST-P positive foci were not decreased as much as in GTI+ group. TBARS contents of carcinogen treated rats decreased by 13weeks of green tea ingestion but GTP groups did not show statiscally significant differences. G6P activities tended to decrease by carcinogen treatment but changes were not statiscally significant by green tea ingestion. Total cytochrome P450 contents were increased by carcinogen treatment. Thirteen weeks of green tea ingestion (GTI) also increased to total cytochrome P450 contents while 7weeks of green tea ingestion(GTP) did show any effects. These results suggest that green tea has suppressive effects on hepatocellular chemical carcinogenesis probably through the activities of antioxidant compounds. (Korean J Community utrition 2(5) : 735∼744, 1997)