• 한국식품위생안전성학회지
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• 제14권1호
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• pp.60-66
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• 1999

Diethylnitrosamine (DEN) is known as a potential hepatic carcinogen by single administration. This study was designed to measure the effects of DEN-induced cell damage on the triglyceride and cholesterol concentration in the liver, excluding dietary effects. Fertilized chicken eggs, 10 days before hatching, were randomly divided into three groups (n=20) and each egg was injected 10 ${mu}ell$ of corn oil (vehicle control), 5 $\mu\textrm{g}$ of DEN/10 ${mu}ell$ of DEN/10 ${mu}ell$ into yolk via air sac. After 48 hr and 96 hr incubation, the damage of the chick-embryo liver cell was investigated by electron microscopy and by measuring the concentration of lipid components (total cholesterol, free cholesterol, phospholipid and triglyceride). For eggs administered 10 $\mu\textrm{g}$ of DEN and incuvated 96 hr, in hepatocyte, the nucleus membrane was roughed, the size of nucleolus was apparently increased and euchromatin was accumulated. Mitochondria were condensed and cristae, located mitochondiral inner membrane, were obscured. Additionally, the leaves of triglyceride and cholesterol classes were significantly increased depend on the amount treated with 10 $\mu\textrm{g}$ DEN at 96 hr, but phospholipids component of cell membrane, were decreased with significance. As a conclusion, carcinogen induced hepatic lesion was correlated with the changes in lipid component of liver.

• Toxicological Research
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• 제27권4호
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• pp.247-251
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• 2011

To clarify whether inhibitory effect of estrogen on liver tumor is associated with cell proliferation, we investigated its role in diethylnitrosamine (DEN)-induced rat preneoplastic lesions, with time sequenced manners. F344 male rats (n = 90) were divided into three groups at 5 weeks of age. The mini-osmotic pumps providing a continuous infusion of DEN was implanted into the abdominal cavity of each animal in group 1, 2 and 3 at 6 weeks of age. To see the effect of estrogen, pellet containing 1 or 10 ${\mu}g$ of estradiol-3-benzoate (EB) was implanted subcutaneously in the animals of groups 2 or 3, respectively, one week prior to DEN treatment. Ten animals of each group were euthanized at 10, 14 and 18 weeks after DEN treatment. Liver tissues at each time point were fixed in 10% phosphate-buffered formalin and were processed and embedded in paraffin and 5 ${\mu}g$ sections mounted on a silanized slide. Glutathione S-transferase placental form (GST-P) positive foci and 5-bromo-2-deoxyuridine (BrdU) labeling cells were detected at each time point. Area of GST-P positive foci in DEN+EB 1 or 10 ${\mu}g$ group was significantly decreased compared to DEN alone at 14 weeks (p < 0.01 or p < 0.05, respectively) an at 18 weeks (p < 0.05 or p < 0.01, respectively). BrdU index in DEN+EB 1 or 10 ${\mu}g$ groups was significantly decreased compared to DEN alone at 14 weeks and at 18 weeks (p < 0.01). Taken together, we conclude that EB treatment decrease the DEN-induced liver preneoplastic lesions and this may be associated with decrease of cellular proliferation.

• 한국환경성돌연변이발암원학회지
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• 제11권2호
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• pp.118-133
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• 1991

신생랫드를 이용한 다장기 발암모델의 전암단계 병변에서 간장의 GST-P 활성도와 발암 과정에 영향을 미치는 phenobarbital (PB)의 발암촉진효과 및 병리조직학적 소견을 관찰하였다. 신생랫드를 150마리 3군으로 나누어 diethylnitrosamine (DEN : 100mg/kg,i,p.), N-me-thyl-N-nitrosourea (MNU : 20mg/kg,i,p.), N-bis(2-hydroxypropyl)nirtosamine(DHPN : 0.1% D. W)을 각각 0, 3, 6 주에 투여하였다. 또한 PB (0.5% in basal diet)를 7주부터 계속 투여하여 8, 12, 20주에 경시적인 부검을 실시하였다.

• Journal of Nutrition and Health
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• 제33권1호
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• pp.23-32
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• 2000

The purpose of this study was to investigate the effcts of n-3, n-6 fatty arid and d-limonene on histopathological and biochemical changes in experimental rat hepatocarcinogenesis. To attain the above objectives, weanling Sprague-Dawley female rats were intraperitoneally injected twice with a dose of diethylnitrosamine(DEN, 50mg/kg body weight) and after 1 week 0.05% phenobarbital was provided with water. Sardine oil rich in n-3 fatty acids and corn oil rich in n-6 fatty acids were fed at 15% by weight and 5% d-limonene was added to the diet in each group. Ten weeks or 20 weeks after DEN treatment, rats were sacrifirced. The formation of glutathione S-transferase placental form positive(GST-P$\^$+/) foci was significantly decreased by the treatment of either sardine oil or d-limonene HMG-CoA reductase activity was not affected by dietary oils and d-limonene. Protein kinase C (PKC) activity was decreased by either sardine oil or d-limonene. Particularly d-limonene decreased the membrane PKC activity. Membrane Cholesterol/Phospholipid(Chol/PL) ratio was significantly decreased by d-limonene in sardine oil group. The data showed that GST-P$\^$+/ foci number was positively correlated with membrane PKC activity and serum cholesterol and negatively correlated with liver cholesterol level. These results suggest informations about the correlation between histopathological and biochemical changes such as cholesterol metabolism and PKC activity in experimental hepatocarcinogenesis and thereby can elucidate the possible mechanism related to the cancer inhibition.(Korean J Nutrition 33(1) : 23-32, 2000)

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1993년도 제2회 신약개발 연구발표회 초록집
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• pp.96-96
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• 1993

간부분 절제술을 하지 않는 비수술적 방법으로서 D-galactosamine을 이용한 중기발암성 시험의 개발을 목적으로 F344 수괵 랫드를 이용하여 본 실험을 수행하였다. 실험 I 에서는 실험방법에 따라 3가지 모델로 구분하고, 각 모델에 처치군과 대조군을 두었다. 모델 1 에서는 실험개시시에 diethylinitrosamine (DEN)을 200 mg/kg body weight로 복강내로 1회 투여하고, 실험개시후 2및 5주에 D-galactosamine을 300 mg/kg body weight로 복강내로 각각 1회 투여하였다. 처치군에는 실험개시후 2주부터 6주간 2-acetylaminofluorene을 0.01%로 혼합한 사료를 급여하였으며, 대조군에는 기초사료를 계속 급여하였다. 모델 2에서는 모델 1의 4주차까지의 처치를 2회 반복하였다. 모델 3은 간부분 절제술을 하는 DEN-PH (diethylnitrosamine-partial hepatectomy) 모델과 같은 방법으로 처치하였다. 사육기간 중 매주 체중 및 사료소비량을 측정하였고, DEN 투여후 8주에 전동물을 부검하여 적출한 간의 중량을 측정하고, glutathione S-transferase placental form (GST-P) 양성 foci에 대한 면역조직화학적 염색표본을 만들어 GST-P 양성 foci의 수 및 면적을 측정하였다. 실험 II에서는 모델 1의 방법으로 phenobarbital(PB), 3-methylcholanthrene (3-MC), n-ethyl-n'-nitro-n-nitrosoguanidine 및 3,3'-diaminobenzidine외 GST-P 양성 foci의 발현정도를 조사하였다. 실험 I의 결과, 모델 1이 정상적인 체중 증가를 보여주었으며, 간조직의 GST-P 양성 foci 의 발현율이 가장 좋았다. GST-P 양성 foci의 면적은 큰것 부터 미상엽, 내측우엽, 외측우엽의 순으로 나타났으나 foci의 수는 모델별로 다르게 나타났다. 실험 II의 PB 투여군과 3-MC 투여군에서 GST-P 양성 foci의 수 및 면적의 유의성 있는 증가가 관찰되었다. 이와 같은 결과로 볼때, 비수술적 방법인 D-galactosamine 을 이용한 중기 발암성 검색법은 간부분 절제술을 이용한 중기발암성 검색법에 비하여 GST-P 양성foci의 발현능력이 동등하거나 더 우수하였으며, 간 및 간이외 장기의 발암물질에 대한 발암성 검색에 보다 유용할 것으로 생각된다.

• 한국수의병리학회지
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• 제5권1호
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• pp.23-28
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• 2001

This study was conducted to assess the chemopreventive effects of sodium selenite in the rat medium-term multi-organ bioassay using a DMBDD model (DEN+MNU+BBN+DMH+DHPN). Seventy five,6-week-old, male SD rats were divided into 3 groups. The animals in group 1 received DEN(diethylnitrosamine,100 mg/kg bw, single i.p., in saline), MNU (N-methyl-nitrosourea,20 mg/kg bw, i.p.,4 times for 2 weeks), BBN (N-butyl-N-(4-hydroxybutyl) nitrosamine, 0.2% in drinking water for 2 weeks), DMH (1,2-dimethylhydrazine, 40 mg/kg bw, s.c., in saline.4 times (or 2 weeds), and DHPN (N-bis(2-hydroxy-pro-pal)nitrosamine,0.1% in drinking water for 2 weeks), then were placed on sodium selenite (4 ppm in drinking water) for 22 weeks from weeks 4 to 26. The animals in group 2 were given DMBDD alone. The animals in group 3 were given sodium selenite alone. Animals were sacrificed at week 12 for ACF quantitative analysis and at week 26 for tumor induction. The body weights in the group 1 were significantly decreased compared with those of group 2. The tumor multiplicities of large intestine in the group 1 were significantly decreased compared with those of group 2 (P<0.05). These results indicate that sodium selenite may have a potential as chemopreventive agents of colon carcinogenesis.

• 한국식품과학회지
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• 제38권4호
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• pp.577-583
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• 2006

유근피 추출물을 20주간 장기 복용한 B6C3F1 마우스의 신장에서 노화 및 생체 독성의 중요한 지표로 알려진 반응성 산소종 발생 및 제거에 관여하는 효소들의 활성 현화를 관찰하였다. 실험군을 1군(대조군), 2군(암유발군-DEN을 주사하여 암유발), 3군(유근피 복용군), 4군(유근피 암치료군-DEN을 주사한 후 유근피복용), 5군(유근피 암예방군-유근피를 먹이면서 DEN 주사)등 5개군으로 나누었다. 암을 유발시킨 군에 유근피 추출물을 투여시 4군 및 5군에서 암유발군에 비해 유의적(p<0.01)으로 감소된 xanthine oxidase 효소활성을 확인하였다. Cu,Zn-SOD의 활성도는 정상군과 처리군에서 큰 차이는 없으나 Mn-SOD의 활성도는 정상군보다 암유발군에서 유의적으로 증가하는 경향을 보였다. 암유발군(2군)과 유근피 암치료군(4군)은 대조군 보다 catalase 효소이 낮았으며, 유근피 암예방군(5군)에서는 암유발군(2군)보다 catalase 활성도가 높은 것으로 나타났다. 암유발군(2군)에서는 glutathione peroxidase의 활성이 감소하는 것으로 나타났으며, 발암물질 처리와 동시에 유근피 추출물을 처리한 유근피 암예방군(5군)에서는 기타 실험군에 비해 효소활성이 매우 증가하는 것으로 (p<0.01) 나타났다. 대조군(1군)에 비해 암유발군(2군)에서 지질의 과산화도는 매우 증가하는 것으로 나타났으며, 유근피 복용군(3군)에서도 유의적으로 증가(p<0.05)하는 경향을 나타내었다. 그러나 암발생 후 유근피 추출물 식이군인 유근피 암치료군(4군)과 암발생과 동시에 유근피 추출물을 식이한 유근피 암예방군(5군)의 경우 암유발군(2군)보다 지질의 산화도가 낮은 것으로 나타났다.

• Molecular & Cellular Toxicology
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• 제4권4호
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• pp.278-284
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• 2008

The X-linked inhibitor of apoptosis (XIAP) is a member of a novel family of inhibitors of apoptosis and has several BIR domains (BIR1, BIR2, and BIR3) and a carboxy-terminal RING zinc-finger. Since suppressionof apoptosis is fundamentally important for carcinogenesis and tumor growth, we investigated the expression and function of XIAP in DEN-induced carcinogenesis using rat model. Wistar rats were injected intraperitoneally with DEN at a dose of 50 mg/kg in twice a week for 12 weeks (Group II) and 16 weeks (Group III) followed by the recovery periods, respectively. The evaluation of DEN-induced carcinogenesis carried out the blood, RT-PCR, histopathological and western blot analysis. The level of blood chemistry including GOT/GPT, albumin, and total bilirubin were significantly exchanged comparing to control and Group I/Group II. The expression of albumin and collagen mRNA were significantly exchanged (P<0.05) in both groups. In addition, AFP mRNA expression decreased more after recovery periods than Group II. XIAP was expressed constitutively in normal rat liver as well as DEN-induced Groups I and Group II. In addition, XIAP expression increased more in Group I with 4 weeks recovery periods than Group I. However, XIAP expression shown to increase in Group lI, otherwise, it was decreased in Group II with 10 weeks repair periods. Taken together, these results suggest the alteration of XIAP expression could be involved in hepatocellular carcinogenesis.

• 한국식품영양학회지
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• 제25권4호
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• pp.1016-1022
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• 2012

In order to investigate the effects of 70% EtOH extract obtained from Hovenia dulcis Thunberg on enzymes relating reactive oxygen intermediate, cancer-stricken animals induced by DEN (N,N-diethylnitrosamine) were recovered by administering the extract of Hovenia dulcis Thunberg. It showed that there was no effect on the generation of superoxide radical by the extract of Hovenia dulcis Thunberg. However, considering the increase of the activity of Cu, Zn-SOD and Mn-SOD in the tested animal class, the extract of Hovenia dulcis Thunberg could participate directly in removing of superoxides. The experimented-animals treated with the extract of Hovenia dulcis Thunberg showed an increase in the activity of the enzymes, catalase and glutathione peroxidase, which can eliminate hydrogen peroxide pertained in liver tissue. The extract of Hovenia dulcis Thunberg seemed to have some factors that accelerate the oxidation. Also, the extract of Hovenia dulcis Thunberg showed effects on the enzymes relating to the active oxygen toxicity which could be an indicator of aging and body toxicity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권1호
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• pp.103-108
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• 2015

Curcumin is widely used as a traditional medicine. This work was aimed to investigate its possible protective effect against chemically induced hepatocellular carcinoma (HCC) in rats. Fifty male albino rats were divided into five groups (n=10, each). The control group received a single dose of normal saline, the diethylnitrosamine (DENA) group received a single intra-peritoneal dose at 200mg/kg body weight, and the 3rd, 4th and 5th groups were given DENA and daily administrated curcunine (CUR) via intra-gastric intubation in doses of 300, 200 and 100 mg/kg b.wt. respectively for 20 weeks. Serum, and liver samples were used for determination of alpha feto-protein (AFP), interleukin-2 (IL-2), interleukine-6 (IL-6), serum liver enzymes (AST, ALT, ALP and GGT) levels as well the activities and gene expression of glutathione peroxidise (GPx), glutathione reductase (GR), catalase (CAT) and super oxide dismutase (SOD). Curcumin significantly lowered the serum levels of AFP, IL-2 and IL-6, ALT, ALT, and malondialdehyde (MDA) as well gene expression of IL-2 and IL-6. In contrast it increased the gene expression and activities of Gpx, GRD, CAT and SOD. The protective effect of CUR against DEN-induced hepatocarcinogenesis in albino rats was proven.