• Applied Biological Chemistry
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• 제38권2호
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• pp.168-173
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• 1995

Endosulfan이 흰쥐(Balb/c.) 체내의 cytochrome P-450 효소계에 미치는 영향을 조사하기 위해 endosulfan을 7.5 mg/kg 수준으로 복강주사하였다. Endosulfan을 복강주사 처리하여 48시간 후 적출한 흰쥐의 간에서는 cytochrome P-450 함량이 $3.3{\sim}4.2$배, cytochrome $b_5$ 함량이 $2.3{\sim}3.8$배, NADPH cytochrome P-450 reductase 활성이 $5.3{\sim}6.4$배 그리고 총 haem 함량이 $3.1{\sim}3.6$배씩 대조구의 그것들에 비해 증가하였다. Endosulfan은 대조구 흰쥐 간의 cytochrome P-450 효소계와 상호작용하여 파장 387와 389 nm에서 흡광도의 증가를 보였으며 파장 407 nm에서 넓은 흡수대를 형성하였다. 환원형 P-450-CO spectrum은 대조구의 경우 파장 451 nm에서 흡광도의 극대를 보인 반면 endosulfan으로 처리된 흰쥐 간의 그것은 파장 449와 450 nm에서 흡광도의 극대를 보였다. Endosulfan 처리로 흰쥐 간과 신장의 aldrin epoxidase 활성이 각각 2.8배와 2.1배씩 증가하였으며 7-ethoxyresorufin dealkylase 활성은 각각 1.7배와 1.8배씩 증가하였다.

• 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.

• Journal of Microbiology and Biotechnology
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• 제22권1호
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• pp.141-146
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• 2012

Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

• 농약과학회지
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• 제9권4호
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• pp.338-346
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• 2005

Carbendazim의 랫드 간 해독체계에 대한 영향을 검색하기 위하여 간장독성의 지표 효소인 혈장 ALT(alanine aminotransferase) 및 AST(aspartate aminotransferase) 활성, 간에 대한 독성, 해독 및 대사에 대한 영향을 구명하기 위하여 간 GSH(glutathione), GST(glutathione-S-transferase), cytochrome P450 및 cytochrome P450 reductase 활성을 375, 750, 1,500 mg/kg 약량에서 측정한 결과 혈장 ALT 및 AST 활성이 120분 후에 약간의 증가가 있었으나 특이한 독성증상은 관찰되지 않았다. GSH는 고농도와 중농도 120분에서 20%의 함량증가가 있었고, GST는 120분까지 $36{\sim}50%$ 정도의 활성저해가 있었으나 240분에는 활성이 회복되었다. Cytochrome P450함량은 60분까지 $25{\sim}50%$까지 함량이 저하되었으나 120분에 70%이상 회복되었고, 240분 저농도에서는 거의 회복되었으며, cytochrome P450 reductase도 120분까지 $25{\sim}50%$ 활성저해가 있었으나, 240분에는 무처리 군의 활성과 유사하게 회복되어 대사관련 효소에 대한 손상은 크지 않을 것으로 판단되었다. 이상의 결과에서 benzimidazole 계 살균제인 carbendazim은 생체내 해독 및 대사관련 체계에 영향이 적은 농약으로 급성적인 중독은 일으키지 않을 것으로 판단되었다.

• 한국식품영양과학회지
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• 제24권6호
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• pp.859-866
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• 1995

본 연구에서는 액체식이로 알코올을 열량의 35%로 6주간 섭취시킨 흰주의 간조직내 지질과산화물과 glutathione 이용효소계의 활성도 및 cytochrome P-450에 미치는 영향을 살펴보고 아울러 간암의 발암원으로 알려진 2-AAF를 투여하여 이들의 상호효과를 조사한 결과 다음과 같은 결과를 얻었다. 1. 체중, 간무게, 그리고 체중에 대한 간무게의 알코올에 대한 효과는 유의적인 차이를 보여 체중은 알코올에 대한 효과는 유의적인 차이를 보여 체중은 알코올 섭취군이 유의적으로 감소하였고 간무게 및 체중에 대한 간무게는 알코올군의 유의적으로 증가하였다. 2. Microsome의 지질과산화물 함량 및 cytosol의 glutathione peroxidase, glutathione reductase 활성도는 알코올과 2-AAF 투여시 모두 유의적인 차이를 보이지 않았으나, cytosol의 glutathione S-transferase 활성도는 알코올과 2-AAF에 의해서 모두 유의적으로 증가하였고 알코올 섭추와 함께 투여시 GST 활성도가 가장 많이 증가하였다. 3. Microsome의 cytochrome P-450 및 cytochrome b5 함량에 대한 알코올 효과는 cytochrome P-450 함량을 증가시키는 경향이 있고 cytochrome b5는 유의적인 증가를 보여 주었으며 2-AAF 투여 역시 cytochrome P-450의 유의적인 증가를 유도하였다. 따라서 알코올 섭취와 2-AAF 함께 투여 시 cytochrome P-450의 함량이 대조군의 약 2.2배 정도 증가하였으며 cytochrome b5 함량이 1.7배로 높이 증가하였다. 이는 2-AAF가 cytochrome P-450을 유도하여 자신의 대사를 촉진시키며 알코올의 섭취 또한 2-AAF의 hydroxylation을 증가시킬 수 있는 것으로 생각된다. 이상의 결과에서 과량의 알코올을 만성적으로 섭취시 간조직내의 microsome의 MFO system에 영향을 미쳐서 발암물질의 생체 활성화를 촉진시킬 수 있고 또한 GST의 활성도를 증가시키므로 어느 정도 발암과정에 영향을 미치는 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제27권5호
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• pp.983-989
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• 2017

NADPH-P450 reductase (NPR) transfers electrons from NADPH to cytochrome P450 and heme oxygenase enzymes to support their catalytic activities. This protein is localized within the endoplasmic reticulum membrane and utilizes FMN, FAD, and NADPH as cofactors. Although NPR is essential toward enabling the biochemical and pharmacological analyses of P450 enzymes, its production as a recombinant purified protein requires a series of tedious efforts and a high cost due to the use of $NADP^+$ in the affinity chromatography process. In the present study, the rat NPR clone containing a $6{\times}$ Histidine-tag (NPR-His) was constructed and heterologously expressed. The NPR-His protein was purified using $Ni^{2+}$-affinity chromatography, and its functional features were characterized. A single band at 78 kDa was observed from SDS-PAGE and the purified protein displayed a maximum absorbance at 455 nm, indicating the presence of an oxidized flavin cofactor. Cytochrome c and nitroblue tetrazolium were reduced by purified NPR-His in an NADPH-dependent manner. The purified NPR-His successfully supported the catalytic activities of human P450 1A2 and 2A6 and fungal CYP52A21, yielding results similar to those obtained using conventional purified rat reductase. This study will facilitate the use of recombinant NPR-His protein in the various fields of P450 research.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.149.3-150
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• 2003

CYP3A4 is the most abundant human CYP and oxidizes a diversity of substrates. including various drugs. steroids. and carcinogens. A variety of metal ions are known to affect microsomal monooxygenase activities. Effects of a series of divalent metal ions on the CYP3A4-catalyzed reaction of reconstituted system containing purified CYP3A4. NADPH-P450 reductase (NPR), and cytochrome b5 (b5) were examined. (omitted)

• Toxicological Research
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• 제11권2호
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• pp.215-221
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• 1995

This paper examines the effects of dietary polyunsaturated fatty acid/saturated fatty acid (p/s) ratios and butylated hydroxytoluene (BHT) on the hepatic microsomaI mixed-function oxidase sy. stem in 2~acetylaminofiuorene (2-AAF) treated rats. Sprague-Dawley male rats were fed the diet of beef tallow (p/s 0.08), beef tallow plus soybean oil (p/s 1.0), and soybean oil (p/s 4.0) at the level of 15%fat and with or without 0.3% BHT. After 2-AAF was injected twice at the ages of 23 and 27 weeks, cholesterol/phospholipid molar ratio, thiobarbituric acid reactive substances (TBARS) level, cytochrome P450, cytochrome $b_5$, NADPH-cytochrome $b_5$, and NADPH-cytochrome c reductase activity were measured from isolated hepatic microsomal fractions. In the beef tallow (p/s 0.08) and beef tallow plus soybean oil (p/s 1.0) groups, cholesterol/phospholipid molar ratio showed decreasing tendency by 2-AAF and BHT. Cytochrome P-450 content was decreased in the group of soybean oil (p/s 4.0) and NADPH-cytochrome c reductase activity was increased by 2-AAF and BHT in all the dietary groups. While TBARS levels were increased by 2-AAF in all the dietary groups, they were reduced by BHT in the soybean oil (p/s 4.0) group. These results suggest that long term intake of soybean oil (p/s 4.0) diet induced changes in the nature of microsomal membrane and induced less cytochrome P-450, low level feeding of BHT increased cytochrome c reductase activity and lowered microsomal lipid peroxidation levels, which were increased by 2-AAF treatment.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.148.2-149
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• 2003

Recently we have reported that various hydroxystilbenes show strong inhibition of human cytochrome P450 1 enzyme activities. A series of syntheic trans-stilbene derivatives were prepared and their inhibitory potentials were evaluated with the bacterial membrane of recombinant human cytochrome P450 1A1, 1A2 and 1B1 coexpressed with hyman NADPH-P450 reductase to find a new inhibitor of cytochrome P450 enzymes. Of the compounds tested, SY-081 exhibited a potent inhibition of human cytochrome P450 1B1 with an $IC_50$ value of 2.6 nM. (omitted)

• 약학회지
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• 제39권4호
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• pp.450-460
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• 1995

The acute toxicity of fthalide in rat was studied in vivo by the observations of the changes in hematogram, serological parameters, content of cytochrome p-450, activities of NADPH-cytochrom c reductase, glucose-6-phosphatase, and the contents of cholinesterase and carboxylesterase in liver. Fthabde is a practically non-toxic substance(LD50 is 3.86g/kg), but rats were intoxicated with fthabde at a oral dose of 100 mg/kg for 12 days. WBC were significantly decreased and activities of ALT and LDH, on the cotrary, the content of glucose in serum were slightly increased. Cytochrome p-450 and lipid peroxide in liver were significantly increased in the fthalide-intoxicated rats. The longer administration of fthalide showed further increase of carboxylesterase activity in liver and serum, but decrease of activities of glucose-6-phosphatase and cholinesterase in liver and serum. These results show that fthatide can induce the hepatocellular injury and neurotoxicity.