• Biomolecules & Therapeutics
• /
• 제20권6호
• /
• pp.562-568
• /
• 2012

Cytochrome P450 BM3 (CYP102A1) from Bacillus megaterium is a self-sufficient monooxygenase that consists of a heme domain and FAD/FMN-containing reductase domain (BMR). In this report, the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) by BMR was evaluated as a method for monitoring BMR activity. The electron transfer proceeds from NADPH to BMR and then to BMR substrates, MTT and CTC. MTT and CTC are monotetrazolium salts that form formazans upon reduction. The reduction of MTT and CTC followed classical Michaelis-Menten kinetics ($k_{cat}=4120\;min^{-1}$, $K_m=77{\mu}M$ for MTT and $k_{cat}=6580\;min^{-1}$, $K_m=51{\mu}M$ for CTC). Our continuous assay using MTT and CTC allows the simple, rapid measurement of BMR activity. The BMR was able to metabolize mitomycin C and doxorubicin, which are anticancer drug substrates for CPR, producing the same metabolites as those produced by CPR. Moreover, the BMR was able to interact with CYP1A2 and transfer electrons to promote the oxidation reactions of substrates by CYP1A2 and CYP2E1 in humans. The results of this study suggest the possibility of the utilization of BMR as a surrogate for mammalian CPR.

• 동의신경정신과학회지
• /
• 제9궈1호
• /
• pp.45-57
• /
• 1998

The effect of HWANSODAN(HSD), on the level of brain antioxidants was examined in aged rat. The experimental groups were divided into three groups and treated as follows ; normal group(negative control), Vt.E administrated Group(HSD). The purified microsome from brain tissue, those were measired the amounts of oxidant materials like Malonfialdehyde(MDA) and H2O2, then activities of antioxidants enzymes like superoxide dismutase, catalase, NADPH-cytochrome P-450 reductase.The results were as follows;1. In TBA reaction to measure the amount of MDA, HSD group and Vt.E group did not showed signigicant decrease.2. In the formation of Hydrogen peroxide, HSD group and Vt.E group showed a little increase.3. The activity of Superoxide dismutase was increased significantly in HSD group and Vt.E group.4. In the activity of Catalase, Vt.E group was increased significantly and HSD group a little increased. 5. The activity of NADPH-cytochrome P-450 reductase in the HSD group and Vt.E group showed significantly increase.According to the above results, it is suggested that HWANSODAN(HSD) has some antioxidant effects on the tissue of brain.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권6호
• /
• pp.2647-2653
• /
• 2012

Involvement of cytochrome P450 genes (CYPs) in breast cancer (BCa) may differ between populations, with expression patterns affected by tumorigenesis. This may have an important role in the metabolism of anticancer drugs and in the progression of cancer. The aim of this study was to determine the mRNA expression patterns of four cytochrome P450 genes (CYP2W1, 3A5, 4F11 and 8A1) in Mexican women with breast cancer. Real-time PCR analyses were conducted on 32 sets of human breast tumors and adjacent non-tumor tissues, as well as 20 normal breast tissues. Expression levels were tested for association with clinical and pathological data of patients. We found higher gene expression of CYP2W1, CYP3A5, CYP4F11 in BCa than in adjacent tissues and only low in normal mammary glands in our Mexican population while CYP8A1 was only expressed in BCa and adjacent tissues. We found that Ki67 protein expression was associated with clinicopathological features as well as with CYP2W1, CYP4F11 and CYP8A1 but not with CYP3A5. The results indicated that breast cancer tissues may be better able to metabolize carcinogens and other xenobiotics to active species than normal or adjacent non-tumor tissues.

• 한국식품영양과학회지
• /
• 제38권10호
• /
• pp.1347-1352
• /
• 2009

본 연구에서는 보리 추출물을 이용하여 알코올 유도 P450 계열 효소 활성을 조절함으로써 라디칼 및 중간체의 발생 감소와 항산화 효소의 활성증강에 의한 보호효과를 연구하였다. ICR mice를 대상으로 대조군(Control), 알코올군(EtOH), 알코올-보리 추출물 급여군(EtOH-B)으로 나누어 전체 열량의 35%에 해당하도록 조성된 알코올 액체 식이를 알코올군 및 알코올-보리 추출물 급여군에 28일 간 공급하였다. 알코올 투여군은 P450 함량에 있어서 대조군에 비하여 급격한 증가를 보였으며, 알코올-보리추출물 급여군은 대조군 수준으로 유의적인 감소를 하였다. 알코올 유도 CYP2E1, CYP1A2는 알코올군에서는 유의적인 증가를 보였으며, 알코올-보리 추출물 투여군에서 이들 효소의 활성을 유의적으로 억제시키는 결과를 확인할 수 있었다. 생체 내 방어시스템의 변화 면에서 알코올 급여군에서 급격하게 감소한 catalase 활성이 보리 추출물 급여군에서는 유의적으로 증가하여, CYP2E1의 활성억제에 의한 라디칼 발생 저하를 확인할 수 있었다. 이상의 실험 결과로서, 보리 추출물은 알코올 유도 CYP2E1, CYP1A2 효소 활성의 억제와 생체 내 방어 기작의 효과적인 활성화에 의하여 알코올성 간 손상의 개선 소재로 활용가능 할 것이라고 사료된다.

• 한국잡초학회지
• /
• 제17권1호
• /
• pp.59-65
• /
• 1997

Bentazon이 첫 분해대사물질인 6-hydroxy bentazon으로 변화하는데 관련하는 cytochrome P-450 활성을 증진시키는 방법을 모색하기 위하여 벼 microsome에서 hydroxylase 효소 유기물질을 처리하여 bentazon 6-hydroxylase(B6H)와 식물에서 흔히 볼 수 있는 cinnamic acid 4-hydroxylase(CA4H) 효소를 대조하여 효소활성을 검정하였다. 1,8-naphthalic anhydride 0.5-2% 농도를 벼 종자에 분의처리하거나 fenclorim 5, 10${\mu}M$을 벼종자에 침지처리함에 따라 B6H와 CA4H 효소 활성이 증대되었다. Ethanol 2.5%를 벼 유묘에 처리함으로써 B6H와 CA4H 활성이 증대되었으며, phenobarbital 12mM 처리에서 B6H 활성이 증대되었고 phenobarbital 2mM 처리에서는 CA4H 활성이 증대되었다. B6H 효소활성은 ethanol 2.5, 5%와 phenobarbital 8, 12mM 혼합 처리 또는 1,8-naphthalic anhydride 0.5, 1%와 phenobarbital 8, 12mM 혼합처리에서 상승적으로 증가하였으며, CA4H 효소는 혼합처리에 의하여 활성이 저하되었다. 한편 벼 5 일묘에서는 B6H와 CA4H 활성 이 가장 높았으며 묘령이 진전될수록 효소활성은 현저히 감소되는 경향을 나타냈다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제5권5호
• /
• pp.407-412
• /
• 2001

Many drugs are primarily metabolized by the cytochrome P450s (CYPs). Drug metabolites would be important allergens for adverse drug reactions such as drug eruptions. Skin tests with a suspected drug have conducted to identify causative drugs of drug eruptions, with vehicles such as white petrolatum, DMSO, ethanol. This study will compare the expression of rat CYP isozyme mRNAs between the skin and the liver, with examining an effect of the vehicles on the cutaneous CYPs using semi-quantitative RT-PCR. Thirty-two Sprague-Dawley rats between the ages of six and eight weeks were divided as four groups. One group was used to compare the constitutive mRNA expression between skin and liver, while the others were to examine the effects of three vehicles. The ratios of expression of CYP1A2, CYP2B1/2, CYP2E1, CYP3A1, and CYP4A1 were significantly higher in the liver than the skin. However, CYP1A1 and CYP2C11 were higher in the skin than liver. The effects of vehicles were quite different; white petrolatum significantly induced CYP1A1 (p=0.012) and CYP2C11 mRNAs, while ethanol inhibited CY P1A1 and CYP2B1/2. DMSO did not make any changes. The results suggest that rat skin can participate in drug metabolism with their own CYP isozymes. The effects of vehicles on the cutaneous CYP expression should not be ignored and may be applied for determination of an appropriate vehicle for certain drug(s).

• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제22권4호
• /
• pp.373-382
• /
• 2000

Individual genetic susceptibilities to cancers may result from several factors including differences in xenobiotics metabolism to chemical carcinogens, DNA repair, altered oncogenes and suppressor genes, and environmental carcinogen exposures. Among them, genetic polymorphisms of metabolizing enzymes to chemical carcinogens have been recognized as a major important host factors in human cancers. They have two main types of enzymes: the phase I cytochrome P-450 mediating enzymes (CYPs) and phase II conjugating enzymes. The purpose of this study is to determine the frequencies of genotypes of phase I (CYP1A1 and CYP2E1) and phase II (NAT2) metabolizing enzymes in healthy control and head and neck cancer patients of Korean and to identify the relative high risk genotypes of these metabolizing enzymes to head and neck cancer in Korean. The author has analyzed 132 head and neck cancer patients and 113 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The results were as following; 1. The frequencies of genotypes of CYP1A1, CYP2E1 and NAT2 in healthy control were as following; CYP1A1 exon 7 polymorphism; Ile/Ile: Ile/Val: Val/Val = 59.3%: 36.3%: 4.4% CYP2E1 Pst I polymorphism, C1/C1: C1/C2: C2/C2 = 61.1%: 32.1%: 6.2% NAT2 polymorphism; F/F: F/S: S/S = 43.4%: 48.7%: 8.0% 2. In analysis of phase I enzyme, Val/Val genotype in CYP1A1 exon 7 polymorphism and C2/C2 genotype in CYP2E1 Pst I polymorphism were associated with relative high risks to head and neck cancers (Odds' ratio: 2.09 and 1.37, respectively). 3. Among the genotypes of NAT2 enzyme polymorphism, S/S genotype of NAT2 enzyme had 1.03 times of relative risk to head and neck cancers. 4. In combined genotyping of CYP1A1, CYP2E1, and NAT2 enzymes polymorphisms, the patients with Val/Val and C1/C1, C2/C2 and fast acetylator, and Val/Val and fast acetylator had higher relative risks than the patients with each baseline of combined genotypes (Odds' ratio: 2.82, 1.98 and 2.1, respectively). These results suggest the combined genotypes of Val/Val and C1/C1, C2/C2 and fast acetylator, and Val/Val and fast acetylator were more susceptible to head and neck cancers in Korean. And genotyping of metabolizing enzymes could be useful for predicting individual susceptibility to head and neck cancer.

• Journal of Microbiology and Biotechnology
• /
• 제22권12호
• /
• pp.1659-1664
• /
• 2012

TSAHC [4'-(p-toluenesulfonylamido)-4-hydroxychalcone] is a promising antitumorigenic chalcone compound, especially against TM4SF5 (four-transmembrane L6 family member 5)-mediated hepatocarcinoma. We evaluated the potential of TSAHC to inhibit the catalytic activities of nine cytochrome P450 isoforms and of P-glycoprotein (P-gp). The abilities of TSAHC to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The P-gp inhibitory effect of TSAHC was assessed by [$^3H$]digoxin accumulation in the LLCPK1-MDR1 cell system. TSAHC strongly inhibited CYP2C8, CYP2C9, and CYP2C19 isoform activities with $K_i$ values of 0.81, 0.076, and $3.45{\mu}M$, respectively. It also enhanced digoxin accumulation in a dose-dependent manner in the LLCPK1-MDR1 cells. These findings indicate that TSAHC has the potential to inhibit CYP2C isoforms and P-gp activities in vitro. TSAHC might be used as a nonspecific inhibitor of CYP2C isoforms based on its negligible inhibitory effect on other P450 isoforms such as CYP1A2, CYP2A6, CYP2B6, CYP2D6, CYP2E1, and CYP3A.

• Archives of Pharmacal Research
• /
• 제27권7호
• /
• pp.693-712
• /
• 2004

The rats with protein-calorie malnutrition (PCM, 5％ casein diet for a period of 4-week) were reported to exhibit 60 and 80％ suppression in the hepatic microsomal cytochrome P450 (CYP) 1 A2 and CYP2C11 levels, respectively, and 40-50％ decreases in CYP2E1 and CYP3A 1/2 levels compared to control (23％ casein diet for a period of 4-week) based on Western blot analysis. In addition, Northern blot analysis showed that CYP1 A2, CYP2E1, CYP2C11, and CYP3A1/2 mRNAs decreased in the state of PCM as well. Hence, pharmacokinetic changes of the drugs in rats with PCM [especially the area under the plasma concentration-time curve from time zero to time infinity (AUC) changes of metabolite(s)] reported from literatures were tried to explain in terms of CYP isozyme changes in the rats. Otherwise, the time-averaged nonrenal clearance ($CL_{NR}$) of parent drug was compared. Pharmacokinetic changes of the drugs in other types of malnutritional state, such as kwashiorkor and marasmus, in both human and animal models were also compared. The drugs reviewed are as follows： diuretics, antibiotics, anticancer agents, antiepileptics, antiarrythmics, analgesics, xanthines, antimalarials, and miscellaneous.

• 한국환경성돌연변이발암원학회지
• /
• 제22권3호
• /
• pp.169-174
• /
• 2002

Resveratrol is known to have potent cancer chemopreventive activity against tumorigenesis caused by 7,12-dimetylbenz［$\alpha$］anthracene(DMBA) which is known to be oxidized to reactive products by cytochrome P450 1B1 (CYP1B1). The effects of resveratrol on the activity of recombinant human P450 1 family enzymes, expressed in Escherichia coli membranes with human NADPH-P450 reductase, were determined by measuring alkoxyresorufin O-dealkylation activity, e.g., ethoxyresorufin O-deethylation (EROD) CYP1A1, methoxyresorufin O-demethylation (MROD), CYP1A2, benzyloxyresorufin-O-debenzylation (BROD), CTP1B1. Resveratrol inhibited CYP1B1 and CYP1A1 activities in a dose-dependent manner with $IC_{50}$/ values of 59 and 10$\mu$M for EROD activity and 1.8 and 30$\mu$M for BROD activity, respectively. Resveratrol had only weak inhibitory effect on CYP1A2 activity ($IC_{50}$/ values of 0.44 mM for EROD and ＞2 mM for MROD). Furthermore, resveratrol did not affect NADPH-P450 reductase activity significantly. Resveratrol inhibited the CYP1B1-dependent EROD activity with a $K_{i}$ of 28 $\mu$M in a non-competitive type manner. these results suggest that resveratrol-derived inhibited of CYP1B1 and CYP1A1 activities may contribute to the suppression of DMBA inducible tumorigenesis observed in extrahepatic tissues.s.