• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.109-109
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• 2001

The present study was done to investigate the relationship between Kupffer cells and alteration of cytochrome P-450 (CYP)-dependent drug metabolizing enzyme activities during polymicrobial sepsis. Male rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP) followed by fluid resuscitation. The gadolinium chloride (GdC1$_3$, 10 mg/kg), blocker of Kupffer cells, was pretreated intravenously at 48 h and 24 h prior to the induction of CLP. All assay parameters were determined at 24 h after CLP or sham operation. In CLP-treated rats, the mortality rate of animals increased to 50% and serum alanine (ALT) and aspartate aminotransferase (AST) levels also significantly elevated. However, this increase was not suppressed by GdC1$_3$ pretreatment. Microsomal lipid peroxidation markedly increased after CLP operation. This increase was significantly attenuated by pretreatment. Total cytochrome P-450 content and NADPH-cytochrome P-450 reductase activity were not changed after CLP operation, but GdC1$_3$pretreatment reduced total cytochrome P-450 content, The hepatic microsomal CYP 1A1, 1A2, 2Bl and 2El activities in CLP-induced rats were also not significantly different from sham-operated rats. However, GdC1$_3$pretreatment showed a moderate increase in CYP1A1 and 1A2 activities. Our findings suggest that Kupffer cells may be partly responsible for producing hepatocellular dysfunction during sepsis.

• 동의생리병리학회지
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• 제22권1호
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• pp.126-130
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• 2008

Water extract from Prunella vulgaris L. (PVW) was tested for colon cancer chemopreventive activity by measuring the activities of cytochrome P450 1A1, phase Ⅱ detoxification enzyme [quinone reductase (QR) and glutathione S-transferase (GST)] and ornithine decarboxylase (ODC) and glutathione (GSH) levels in cultured human colorectal adenocarcinoma HT-29 cells. PVW significantly inhibited 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P450 1A1 activity at 10 and 50 ${\mu}g/ml$. PVW induced QR activity in a dose-dependent manner over a concentration range of $1{\sim}50\;{\mu}g/ml$. GST activity was also induced with the treatment of PVW in HT-29 cells. In addition GSH levels were increased with PVW. PVW inhibited ODC activity, a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. These results suggest that Prunella vulgaris L. has colon cancer chemopreventive activity by inhibiting cytochrome P450 1A1 and ODC activities and by increasing phase Ⅱ enzyme activity and GSH levels.

• 한국균학회지
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• 제31권1호
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• pp.34-39
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• 2003

싸리버섯 메탄올 추출물이 간독성 물질인 B$({\alpha})$P을 투여한 마우스에서 간 손상 억제 효과 및 cytochrome P-450 1A1 발현에 미치는 영향을 살펴보았다. B$({\alpha})$P 투여로 인한 혈청 ALT와 AST의 활성, 간 조직증의 lipid peroxide 함량, cytochrome P-450 함량, AD 및 AH 활성이 유의적으로 증가하였으며 싸리버섯 메탄올 추출물의 투여시 이들 활성 및 함량이 유의적으로 감소하였다. 반면, 간 조직중의 GSH 함량, GST, r-glutamylcysteine synthetase의 활성은 B$({\alpha})$P만 투여한 군에 비해 싸리버섯 메탄올 추출물을 투여시 증가하였다. 또한 immuno blotting 결과로부터 B$({\alpha})$P 투여에 의해 현저히 증가되었던 cytochrome P-450 1A1 isozyme 단백질 함량이 싸리버섯 메탄올 추출물의 투여로 감소됨을 확인하였다. 이와 같은 결과로부터 싸리버섯 메탄올 추출물은 B$({\alpha})$P에 의한 간 손상에 대한 보호 효과가 있는 것으로 사료된다.

• 약학회지
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• 제34권2호
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• pp.69-79
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• 1990

The aim of this experiment is to observe the toxicity of cypermethrin[S, R- -cyano-3-phenoxybenzyl-(1R, 1s, cis, trans)-2,2-dimethyl-3-(2,2-dichlorovinyl) cyclopropane carboxylate]and to investigate the synergistic effect of piperonyl butoxide on the cypermethrin toxicity. In cypermethrin (CYP) treated group, the biochemical parameters such as ALT, LDH, glucose in serum were remarkably elevated. The content of cytochrome P-450 and activity of NADPH-cytochrome c reductase in renal microsomal fraction were increased but those in hepatic microsomal fraction were not significantly increased. The activity of aniline hydroxylase and ATPase in liver were decreased. In the case of CYP plus piperonyl butoxide (PB) treated group, AST, ALT, LDH and glucose were more increased. Cytochrome P-450 and NADPH-cytochrome c reductase in liver and kidney were supressed and aniline hydroxylase and ATPase in liver were more decreased. Especially, in the case of CYP plus PB 100 mg/kg treated group, hepatic TBA value was increased but activity of glucose-6-phosphatase was remarkably depressed.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Signal transduction in Toxicology
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• pp.143-143
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• 2001

Human cytochrome P450 (CYP or P450) 3A4 (CYP3A4) is the most abundant among P450s in human liver. We previously reported that the expression of CYP3A4 in membranes prepared from E. coli coexpressed the bicistronic construct of CYP3A4 and NADPH-P450 reductase with cytochrome b$_{*}$ (b5) was showed 20-60% higher than that in membranes from E. coli expressed only the bicistronic construct with culturing longer times (48-72h).(omitted)

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.201-205
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• 2002

It is generally accepted that the immune system is one of the major target organs for many toxic chemicals. In addition, many toxic chemicals require metabolic activation by cytochrome P450s for their toxicity. Although the immune cells possess a limited amount of drug metabolizing capacity, metabolic activation of certain toxicants in liver and immune organs may have a significant role in immunosuppression. In the present studies, the possible role of metabolic activation by cytochrome P450s in chemical-induced immunosuppression was reviewed, with a particular emphasis on the methodological techniques to detect immunotoxicants requiring metabolic activation in vivo and in vitro. (omitted)

• 생명과학회지
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• 제19권8호
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• pp.1039-1046
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• 2009

다양한 천연물의 합성대사에 관여하는 식물 cytochrome P450 (P450s)은 그 기능적 다양성에도 불구하고, 이들 효소의 광범위한 기질 특이성을 설명해 줄 수 있는 구조분석에 대해서는 충분한 연구가 이루어지지 못하고 있는 실정이다. 식물 p-coumarate 3-hydroxylase (C3H)에 의해 매개되는 효소 반응은 lignin 과 다양한 phenylpropanoid 부산물들의 생합성에 매우 중요한 것으로 여겨지지만, 막 단백질인 C3H의 발현 및 정제가 효과적으로 이루어지지 못하여, 활성을 측정하기 위한 분석방법이 체계화 되지 못하고 있다. C3H의 작용기작과 기질특이성에 대해 폭넓은 이해를 위한 구조분석의 선행단계는 활성을 갖는 C3H를 밀리그램 단위로 분리, 정제하는 실험적 방법을 확립하는 것이라 할 수 있다. 이를 위해, 본 연구에서는 다양한 돌연변이 방법을 도입하여 식물 막단백질 C3H를 대장균 시스템에서 효과적으로 발현 및 정제할 수 있는 시스템을 사용하였다. 변형된 cytochrome P450 C3H ($C3H_{mod}$)을 세포막으로부터 고농도의 염완충용액을 이용하여 계면활성제 없이 추출하였으며, 2단계 chromatography를 통해 활성을 유지한 상태로 분리할 수 있었다. 이러한 실험적 기법은 NMR 및 X-ray crystallography와 같은 구조분석을 통한 C3H의 효과적인 분석에 적용될 수 있을 것이며, 또한 다른 식물 cytochrome P450 단백질의 효과적인 분석에도 적용 될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1011-1017
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• 2001

Streptomyces sp. Y-110 cytochrome P-450, induced by the addition of compactin -Na into the culture medium, was purified from the cell extract to apparent homogeniety, mainly by DEAE-Sepharose, hydroxyapatite, and Mono Q column chromatyography. The sepcific activity of purified enzyme on its substrate, compactin-Na, was determined to be 15 nmol of pravastatin per mg protein. The molecular mass of this enzyme on SDS-PAGE was $37{\pm}0.5$ kDa, pI was 4.5, and its CO difference spectrum showed maximum absorption peaks at 452 and 550nm, respectively. The N-terminal amino acid sequence was determined to be Met>Thr>Cys>Thr>Pro>Val>Thr>Val>The>Gly>Ala>Ala>Gly>Gln>Ile>Gly>Tyr>Ala>Leu. Its apparent $K_m$ on compactin-Na was $1.294{\mu}M{\cdot}min^-1,\;and\;V_{max}\;was\;1.028{\mu}M{\cdot}min^-1$. The maximum substrate concentration ($K_s$) for reaction was $270 {\mu}M$and thus $1/[K_s]$ was $3.7{\mu}M$. These physicochemical characteristics and kinetic behavior of this enzyme were compared and shown to be different from those of Streptomyces cytochrome P-450 enzymes reported, suggesting that this enzyme may be an additional member of the Streptomyces cytochrome P-450 family.

• Environmental Analysis Health and Toxicology
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• 제7권3_4호
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• pp.57-67
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• 1992

BPMC (2-Sec-butylphenyl N-methylcarbamate) was treated at the level of 100mg/kg/day in oral administration for 12th days in rat. It was investigated not only that the hematogram and the serological parameters, but also the content of cytochrome P-450, the activity of TBA, glucose-6-phosphatase, cholinesterase and carboxylesterase in rat. The results were as follows: The hematogram was not found any alteration but the value of AST, ALT, LDH and the content of glucose in serum were significantly increased compare with that of control group. The content of cytochrome P-450 in liver was increased significantly on the contrary cytochrome P-450 in kideny and NADPH-cytochrome c reductase in liver and Kidney were not significantly increased. After the final 12th day, the value of TBA and the activity of glucose-6-phosphatase appeared to the tendency of increasement in the liver. The activity of cholinesterase and carboxylesterase both in serum and liver were decreased. Especially the activity of cholinesterase was more significantly decreased. It was conclusion that the function of this insectivide should be due th the inhibition of cholinesterase activity.

• Journal of Plant Biotechnology
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• 제43권4호
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• pp.422-431
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• 2016

Plants utilize cytochrome P450, a large superfamily of heme-containing mono-oxygenases, in the synthesis of lignins, UV protectants, pigments, defense compounds, fatty acids, hormones, and signaling molecules. Despite the overwhelming assortment of rice P450 accession numbers in the database, their functional studies are lacking. So far, there is no evidence involving rice P450 in disease immunity. Most of our understanding has been based on other plant systems that are mostly dicot. In this study, we isolated the cytochrome P450 (OsCYP71) in rice, and screened the gene using gain-of-function technique. The full-length cDNA of OsCYP71 was constitutively overexpressed using the 35S promoter. We then explored the functions of OsCYP71 in the rice - Xanthomonas oryzae pv. oryzae pathosystem. Using the gene expression assays, we demonstrate the interesting correlation of PR gene activation and the magnitude of resistance in P450-mediated immunity.