• Food Science and Biotechnology
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• 제15권6호
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• pp.942-947
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• 2006

The effect of oat bran extracts on the formation of aberrant crypt foci (ACF) in the colon induced by 1,2-dimethylhydrazine (DMH) was studied in F344 male rats. Extracts were prepared using various combinations of temperature (40, 45, 50, 55, or 60$^{\circ}C:\;X_1$), ethanol concentration (0,5, 10, 15, or 20%: $X_2$), and pH (5, 6, 7, 8, or 9: $X_3$). Among the various extracts tested, one ethanol extract (EE; $45^{\circ}C$, 15% ethanol at pH 6) and one water extract (WE; $50^{\circ}C$ at pH 5) were selected based on their in vitro antitumor activity. The animals were fed with basal diet alone or basal diet supplemented with 0.25 or 0.5% of EE or WE for 6 weeks. During the initial 2 weeks of the 6-week test period, the rats were subcutaneously injected with DMH (30 mg/kg) 4 times for the induction of ACF. DMH induced an average of 322.7 and 142.9 aberrant crypts (AC) and ACF, respectively. A low dose (0.25%) of EE (containing 38.3% ${\beta}$-glucan) and WE (containing 22.8% ${\beta}$-glucan) greatly reduced the numbers of DMH-induced AC and ACF. Significantly, ACF consisting of more than 3 AC were reduced by half in which the effect of EE, containing a higher concentration of ${\beta}$-glucan, was superior to that of WE. These results demonstrate that oat bran extracts may confer protection against colon carcinogenesis.

• Molecular & Cellular Toxicology
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• 제1권4호
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• pp.256-261
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• 2005

The recent DNA microarray technology enables us to understand gene expression profiling in cell line and animal models. The technology has potential possibility to comprehend mechanism of multiple genes were related to compounds which have toxicity in biological system. So, microarray system has been used for the prediction of toxicity through gene expression induced by toxicants. It has been shown that compounds with similar toxic mechanisms produce similar changes in gene expression in vivo system. Here we focus on the use of toxicogenomics for the determination of gene expression analysis associated with hepatotoxicity in rat liver and cell line (WB-F344). Methotrexate (MTX) is a chemotherapy agent that has been used for many years in the treatment of cancer because it affects cells that are rapidly dividing. Also it has been known the toxicity of MTX, in a MTX abortion, it stops embryonic cells from dividing and multiplying and is a non-surgical method of ending pregnancy in its early stages. We have shown DNA microarray analyses to assess MTX-specific expression profiles in vivo and in vitro. Male Sprague-Dawely VAF+ albino rats of 5-6 weeks old and WB-F344 cell line have been treated with MTX. Total RNA was isolated from Rat liver and cell line that has treated with MTX. 4.8 K cDNA microarray in house has been used for gene expression profiling of MTX treatment. We have found quite distinct gene expression patterns induced by MTX in a cell line and in vivo system.

• 생명과학회지
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• 제15권1호
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• pp.87-93
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• 2005

본 연구에서는 Streptozotocin-유발 당뇨 쥐를 대상으로 8주 동안 운동 강도에 따른 혈당, 인슐린, 골격근 내 GLUT-4 발현 량과 GRP-78 발현량 변화를 관찰함으로써 당뇨 쥐의 혈당을 낮추는데 효과적인 운동 강도는 무엇이며, 기전적인 원인은 무엇인지를 밝히고자 실시되었다. 본 연구에서 Streptozotocin으로 당뇨를 유발시킨 흰쥐를 대상으로 운동 강도 처치에 따른 GLUT-4발현 량을 비교한 결과, 기본적으로 당뇨가 유발된 흰쥐의 GLUT-4 발현 량은 정상 쥐에 비해 현저하게 감소하는 것을 볼 수 있었다. 이중 8주 동안 저강도 운동 수행 한 쥐들의 경우는 정상 쥐 수준에 가까운 발현 량을 관찰 할 수 있었으며, 흰쥐 가자미 근 내 GRP-78 발현 량은 당뇨 유발 쥐의 경우 정상 쥐의 유의하게 증가한 것을 관찰할 수 있었고, 저강도 운동을 수행한 집단에서의 GRP-78 발현 량은 정상 흰쥐에서 발현되는 양에 비해 유의하게 높은 수준을 보이기는 했지만 당뇨 유발 대조군과 고강도 운동 군에 비해서는 유의하게 낮은 수준으로 나타났다. 결론적으로 저강도의 운동은 당뇨 질환자의 혈당을 낮추는 데 가장 효과적인 운동이라 할 수 있으며, 이러한 결과의 기전적인 원인은 저강도의 운동이 근육 내 GLUT-4의 증가로 세포내 당 유입을 촉진시킴으로 해서 나타난 사실이라 생각되며 이 같은 사실은 GRP-78의 발현 량의 감소로 확인할 수 있었다.

• 환경생물
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• 제24권2호
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• pp.102-111
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• 2006

KWNP의 특정 손상에 대한 회복 효과에 대한 과학적인 접근을 하기 위해서 본 연구를 수행하였다. 실험은 STZ를 복강 주사하여 당뇨를 유발, $220mg\;dL^{-1}$ 이상인 실험동물을 이용하여 수행되었으며 방사선 전신 조사 실험군과 KWNP 처리군 간의 각 항목 분석치를 비교, 분석하였다. 실험 5주간의 체중증가율은 방사선 처리군과 당뇨군 모두 대조군에 비하여 낮게 나타났으며 특히 당뇨군에서는 통계적으로 유의적으로 낮았다. 정소와 비장, 또 당뇨병의 직접적인 병증이 나타나는 신장의 무게를 비교한 결과, 방사선을 조사한 실험군의 경우 대조군과 비교하여 정소의 무게의 감소는 비교적 뚜렷하게 관찰된 반면 비장의 무게는 유의한 감소가 관찰되지 않았다. 당뇨군에서는 대조군과 비교하여 정소의 현저한 무게감소가 관찰되었으며 신장의 무게는 유의하게 증가되었다. 혈액 내 지표인자의 수준 변화는 적혈구의 경우 모든 실험군에서 감소하는 경향을 보였으며, 백혈구의 경우에는 방사선처리군, STZ와 방사선 복합 처리군, 당뇨군에서 낮은 수치를 나타냈다. ALP의 측정 결과, 당뇨군에서 그 수치가 현저하게 높아짐을 확인할 수 있었고, STZ와 방사선, KWNP를 복합 처리한 실험군에서는 백혈구의 수치 및 ALP 수치를 측정한 결과 모두에서 방사선 처리구나 당뇨군에 비하여 그 회복능이 현저히 높은 것을 확인할 수 있었다. 혈중중성지질의 증가율 확인결과 KWNP를 처리한 모든 군에서 현저한 지질의 감소를 확인 할 수 있었다. 인슐린의 수치 측정 결과, STZ와 방사선, KWNP를 복합 처리한 실험군에서 다른 당뇨군과 비교하여 10배 정도 높은 insulin수치가 측정되었다. 조직학적 검경시 정소의 경우, 방사선 처리군과 당뇨군 모두에서 정소 세정관의 직경감소와 세정관 내부의 공포화가 관찰되었고 당뇨군의 경우 세정관 내부의 비가 역적 손상이 관찰되었다. 신장의 경우 유의한 형태적 차이는 관찰되지 않았으나 당뇨군의 근위곱슬세관 분분에 세포 자연사가 중점적으로 발견되었다.

• Toxicological Research
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• 제21권2호
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• pp.141-150
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• 2005

1,3-Dichloro-2-propanol (1,3-DCP) is known as chloride chemicals and causes severe hepatotoxic agent. The Ito cells and Kupffer's cells of the liver in the 5 old F344 Rats were exposed to 1,3-DCP gas chamber for 6 hours/ a day, 5 days/ a week, and 13 weeks, in the 0, 5, 20, 80 ppm, respectively. After then the body weights, liver weights, and relative liver weight to body weight were measured, and the hepatic tissues were prepared by the routine and Immunostain method, and observed by the LM, and EM. In the results, there were severe body weight decrease (p<0.05) in the 80 ppm of the male and female rats. The relative liver weights to the body weight were increased relate with exposed 1,3-DCP concentration (P<0.001). Inflammatory cells, infiltration was observed at the perivascular area in the 20 ppm exposed group, and bilirubin pigment infiltration, bile duct hyperplasia, inflammation hepatocytic necrosis, fibrosis were observed in the 80 ppm exposure group. In the 80 ppm exposure group, disarrangement of the endothelial cells, erythrocytes and hepatic cell fragment in the Disse space and numerous migration macrophages were observed in the necrotic area by EM observation. In the immunostained hepatic tissues positive stained ED1 cells were extremely increased (P<0.05) in central vein area, but ED2 was weakly positive immunostained in the 80 ppm exposed group. Immunostained desmin was observed in the Ito cell. It was no difference in the low and medium exposed group but it was typical increase in the necrotic area. In conclusion, These results suggest that NOAEL of 1,3-DCP may be 5 ppm in rats and the Immunostained of desmin, ED1 and ED2 positive cells activated in the inflammatory liver were related to the exposure volume and density. The increase of the Ito cells were related to the severe phagocytosis of the Kupffer's cells.

• Toxicological Research
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• 제23권4호
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• pp.321-330
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• 2007

Garlic (Allium sativum L.) with the food supplement material and medicine was used traditionally in Asia and Europe. Epidemiological studies revealed that the intake of garlic reduced incidences of various cancer including digestive system. The present study was designed to investigate the effect of garlic ethanol extract on the development of colonic aberrant crypt foci (ACF) induced by 1,2-dimethylhydrazine (DMH) in male F344 rats. Five-week-old rats were given four times for two weeks to subcutaneous injections by DMH (30 mg/kg body weight) to induce ACF. The animals were divided into groups that fed diet containing garlic ethanol extract at five different doses (0.1, 0.2, 0.5, 2, 5%), respectively, animals were evaluated for the total number of ACF and total aberrant crypts (AC) per colon detected from methylene blue-stained rat colon. ACF were formed in animals in DMH-treated group. The feeding suppressed potently the appearance ACF in the colon of rats. Especially, fed diet containing garlic ethanol extract at intermediate dose (0.5%) significantly reduced the number of ACF and AC per colon (p < 0.05). Garlic ethanol extract inhibited DMH-induced overexpression of Activator Protein-1 (AP-1) and ${\beta}-catenin$ genes related to cell proliferation that also upregulated the expression of p21Waf1/Cip1 mRNA, a cell cycle-regulating gene. These results suggested that garlic ethanol extract may inhibit ACF formation, ${\beta}-catenin$ gene as the early preneoplastic marker of malignant potential in the process of colon carcinogenesis.

• 약학회지
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• 제57권4호
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• pp.258-264
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• 2013

Herbal medicine has been traditionally used in Asian countries for a long time. Many pharmacological effects are identified in the herbs and these herbs are believed to be safe for human. However, the safety or adverse effect of some traditional herbal medicines has not been established. We have chosen Cyperi rhizoma based on the Korea Herbal Pharmacopoeia and which have been widely used for an anti-inflammatory effect in Korea. The object of the study was to evaluate safety of Cyperi rhizoma in rats. The aqueous extract of Cyperi rhizoma was prepared according to the standard hot water extraction method of the Korea Pharmacopoeia. In the sub-chronic study, the aqueous extract of Cyperi rhizoma was orally administered once daily as 0, 125, 250, 500, 1000 and 2000 mg/kg/day to male and female F344 rats for 13 weeks. There were no treatment related abnormalities in mortality, clinical signs, food consumption, ophthalmologic examination, hematology, serum chemistry, urinalysis, gross observation, organ weight and histopathologic examination. In conclusion, The NOAEL (No Observed Adverse Effect Level) for Cyperi rhizoma aqueous extract was determined as more than 2000 mg/kg/day in the present experimental condition.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1286-1294
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• 2004

In a previous study by the current authors, hepatocellular carcinoma (HCC) was determined to be epidemiologically sex-dependent, and the incidence and multiplicity of HCC found to decrease in estradiol-3 benzoate (EB)-treated F344 rats. Therefore, to ascertain the anticancer mechanism of EB, a commercially available cDNA microarray, with a total of 14,815 cDNA rat gene clones, was used to determine the differentially expressed genes in nontreated livers, EB-treated livers, and diethynitrosolamine (DEN)-induced HCC. In the sequenced experiment, a total of 85 genes were differentially expressed at either two or more times the rate of the normal expression, where 33 genes were downregulated by EB, and 52 genes upregulated. Candidate genes were selected according to significant changes observed in the mRNA expression in the EB-treated livers compared with the nontreated livers, then these genes were filtered according to their different expression patterns in the DEN-induced tumors compared to the estrogen-treated livers. To confirm the microarray data, a real-time PCR analysis was performed for ten selected genes: the H-ras revertant protein 107 (H­rev107), insulin-like growth factor binding protein (lOFBP), parathyroid hormone receptor (PI'HR), SH3 domain binding protein (SH3BP), metallothionein, src-suppressed C-kinase substrate (SSeCK) gene, phosphodiesterase I, CD44, epithelial membrane protein 3 (EMP3), and estrogen receptor a (ERa). The SSeCK and phosphodiesterase I genes were both upregulated in the DEN-induced hepatocarcinomas, yet their possible carcinogenic functions remain unknown. Meanwhile, the other genes were downregulated, including the genes related to growth regulation (IOFBP, H-revI07, ER$\alpha$), adipogenesis inhibition (PTHR), and tumor suppression (metallothionein).

• Archives of Pharmacal Research
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• 제20권4호
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• pp.297-305
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• 1997

A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

• 한국수의병리학회지
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• 제5권1호
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• pp.29-34
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• 2001

This study was conducted to assess the chemopreventive effects of chitosan in a rat colon carcinogenesis induced by azoxymethane (AOM). Ninety, 5-week-old, male F344 rats were divided into three groups. The animals in group 1 received subcutaneous injections of 15mg/kg AOM three times for two weeks, then were placed on powdered basal diet containing 2% chitosan for 37 weeks from weeks 3 to 40. The animals in group 2 were given AOM alone. The animals in group 3 were given 2% chitosan without prior carcinogen treatment. All animals were sacrificed at week 12 for quantitative analysis of aberrant crypt foci (ACF) and at week 40 fur analysis of tumor induction. Total numbers of ACF and AC per colon of group 1 were not significantly different from those of group 2. Tumor incidences and multiplicities of small intestine in the group 1 were significantly decreased compared with those of the group 2 (P<0.05). According to pathological diagnoses, adenocarcinoma incidence and multiplicity in the small and large intestine in the group 1 were significantly decreased compared with those of the group 2 (p<0.05). No toxic effects were observed in animals given chitosan in terms of body weights, and liver or kidney histology. These results indicate that chitosan may have a potential as chemopreventive agents of colon carcinogenesis during the postinitiation stage.