• Environmental Analysis Health and Toxicology
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• 제20권3호
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• pp.195-203
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• 2005

Cytochrome P45O 17$\alpha$-hydroxylase (CYPI 7) and cytorhrome P45O aromata.ie (CYPI 9) are key steroidogenic enzymes in androgen and estrogen synthesis. ThiL study evaluated the effects of nonylphenol (NP) on CYP17 and CYP19 expression in the ovary of Sprague-Dawley rats. All female rats were administered orally with the vehicle (control, corn oil), diethylstilbestrol (DES, 5.0 $\mu$g/kg) and NP (50, 100, or 200 mg/kg/day), which was startinB when they were weaned at 21 days of age for 20 days. Twenty four hours after final dose, the animals were anelthetized with ether. Significant decreases in the uterus (wet weight) were observed with 5.0 $\mu$g/kg/day DES (78$\%$, of control) and 200 mg/kg/day NP (62$\%$ of control), respectively Additionally, ovarian weight was significantly decreased with 5.0 $\mu$g/kg/day DES (63$\%$ of control) and 200 mg/kg/day NP (72$\%$ of control). The serum estradiol levels were sligHtly lower in DES and high dose NP treatment groups, but the 74 levels were not affected by DES and NP. The expression of the ovarian CYP19 gene increased with low doses (50 and 100 mg/kg/day) of NP. while DES and high dose oi NP (200 mg/kg/day) did not affect on the CYP19 mRNA levels. In contrast to the CYP19 gene, the CYP17 gene expreLsion level was significantly down-regulated by the DES and 200 mg/ks/day NP. This result suggestE that NP inhibits ovarian estrogen synthelis by supprelsing CYP17 mRNA efprelsion, And different mechanisml might exist for the expression of Lteroidogenic CYP17 and CYP19 genes in the ovary of Sprague-Dawley rats in response to NP.

• Mass Spectrometry Letters
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• 제12권4호
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• pp.172-178
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• 2021

HYIpro-3-1 is an adjuvant for preventing or treating inflammatory growth diseases. In this study, we identified the metabolic pathway of HYIpro-3-1 in human liver microsomes (HLMs) by quadrupole-orbitrap high-resolution mass spectrometry (HR-MS) and characterized the major human cytochrome P450 (CYP). Ten metabolites were identified, including one O-demethylation (M1), two O-demethylation and monohydroxylation (M2 and M3), and seven monohydroxylation metabolites (M4-M10). Based on the HR-MS² spectra, the metabolites are divided into two groups of monohydroxylated metabolites according to the hydroxylation position. We verified that HYIpro-3-1 is metabolized by CYP in HLMs, CYP2B6 is mainly involved in O-demethylation, and various CYPs are involved in the monohydroxylation of HYIpro-3-1.

• Clinical and Experimental Pediatrics
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• 제48권4호
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• pp.380-386
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• 2005

목 적 : 신생아 황달은 백인에 비해 중국, 일본, 한국 등 동아시아인에서 2배 이상 많이 발생하는 것으로 보아 유전적 연관성이 있을 것으로 생각되어 왔고, 최근 일본인, 타이완 중국인, 한국인에서 UGT1A1 유전자의 Gly71Arg 다형성이 신생아 황달과 연관성이 있다고 보고되었다. 선천적으로 UDP-glucuronosyltransferase(UDPGT)에 결함이 있는 경우에는 부경로(alternative pathway)로 CYP1A2 효소를 자극하여 빌리루빈 대사가 이루어진다. 출생 후 6-14주가 되어야 성인 UDPGT 정상치에 도달하기 때문에 신생아 황달에서 빌리루빈 대사에 CYP1A2 부경로가 중요한 역할을 할 것으로 생각된다. 이에 저자들은 UGT1A1과 CYP1A2 유전자의 다형성이 한국인 신생아 황달의 발생과 어떤 연관성이 있는지 알아보고자 본 연구를 시행하였다. 방 법 : 혈중 빌리루빈 수치가 12 mg/dL 이상의 건강하고, 황달의 다른 위험인자가 없는 만삭아 79명과 대조군 68명으로부터 혈액 0.5 cc를 채취하여 DNA을 분리하였다. UGT1A1 유전자는 Polymerase chain reaction(PCR) 후에 염기서열 분석을 통해서 Gly71Arg 유전자 다형성을 확인하였으며, CYP1A2는 제한효소인 MboII를 이용하여 PCR-restriction fragment length polymorphism 방법과 염기서열 분석을 통해서 T2698G 유전자 다형성을 확인하였다. 결 과 : UGT1A1 유전자의 Gly71Arg 다형성은 변이형 대립 유전자 분포가 환자군에서 32%로 대조군 11%보다 높았다(P<0.0001). CYP1A2 유전자의 다형성은 변이형 유전형 분포가 환자군에서는 41.8%, 대조군에서 32.3%로 환자군이 높았으며 통계학적으로 유의하였다(P=0.015). 변이형 대립유전자의 빈도는 환자군에서 21%로 대조군 19%보다 높았으나 통계학적 유의성은 없었다(P=0.706). Gly71Arg와 T2698G의 변이형 발생의 연관성은 없었다(P=0.635). 결 론 : 한국인의 신생아 황달에서 체내의 빌리루빈 대사의 주경로와 부경로에 작용하는 효소의 유전자인 UGT1A1과 CYP1A2의 다형성이 확인되었고, UGT1A1 유전자의 Gly71Arg 다형성은 신생아 황달과 연관이 있었으나 CYP1A2 유전자의 T2698G 다형성은 신생아의 황달과 연관이 없었다.

• 한국산업보건학회지
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• 제21권1호
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• pp.49-54
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• 2011

In order to develop the methods for exposure assessment and find susceptibility markers for the workers who are exposed to low doses of toluene, xylene and other chemical in petroleum industries, we investigated the application of P-450 expression in human lymphocytes utilizing mouse monoclonal anti-rat CYP2B1/2, the levels of toluene and xylene in air and their metabolite levels in urine with the levels of expressed CYP2B1/2 proteins. The general characteristics such as age, smoking and drinking habit were no significant difference between the control and exposed workers, but the working durations and working hours were significantly different. Workers in exposed group were exposed to the mean of 2.1 ppm (range, 0.00-4.54) of toluene and 0.3 ppm (rang, 0.00-1.23) of xylene. The mean concentration of urinary hippuric acid was low and less than 1/5 of the biological exposure index recommended by the Ministry of Employment and Labor Korea. Methyl hippuric acid in urine was not detected in control and exposed workers. Also, there were no significant differences in the levels of the urinary metabolites between the control and exposed group. When chemiluminescence dot blottings were carried out utilizing mouse monoclonal antibody against CYP2B1/2, the strong density dots corresponding to a mouse monoclonal antibody was observed in the human lymphocytes from the exposed workers. These results suggested that the chemiluminescence dot blot assay for CYP of lymphocytes should be valuable for identifying CYP expression as biomarkers in the workers exposed to toluene and xylene.

• Biomolecules & Therapeutics
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• 제21권6호
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• pp.487-492
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• 2013

Cytochrome P450 4A11 (CYP4A11) is a fatty acid hydroxylase enzyme expressed in human liver. It catalyzes not only the hydroxylation of saturated and unsaturated fatty acids, but the conversion of arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE), a regulator of blood pressure. In this study, we performed a directed evolution analysis of CYP4A11 using the luminogenic assay system. A random mutant library of CYP4A11, in which mutations were made throughout the entire coding region, was screened with luciferase activity to detect the demethylation of luciferin-4A (2-[6-methoxyquinolin-2-yl]-4,5-dihydrothiazole-4-carboxylic acid) of CYP4A11 mutants in Escherichia coli. Consecutive rounds of random mutagenesis and screening yielded three improved CYP4A11 mutants, CP2600 (A24T/T263A), CP2601 (T263A), and CP2616 (A24T/T263A/V430E) with ~3-fold increase in whole cells and >10-fold increase in purified proteins on the luminescence assay. However, the steady state kinetic analysis for lauric acid hydroxylation showed the significant reductions in enzymatic activities in all three mutants. A mutant, CP2600, showed a 51% decrease in catalytic efficiency ($k_{cat}/K_m$) for lauric acid hydroxylation mainly due to an increase in $K_m$. CP2601 and CP2616 showed much greater reductions (>75%) in the catalytic efficiency due to both a decrease in $k_{cat}$ and an increase in Km. These decreased catalytic activities of CP2601 and CP2616 can be partially attributed to the changes in substrate affinities. These results suggest that the enzymatic activities of CYP4A11 mutants selected from directed evolution using a luminogenic P450 substrate may not demonstrate a direct correlation with the hydroxylation activities of lauric acid.

• Journal of Microbiology and Biotechnology
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• 제21권1호
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• pp.14-19
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• 2011

A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the $4^{th}$ N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana${\Delta}$CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.

• Korean Journal of Acupuncture
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• 제21권2호
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• pp.139-145
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• 2004

Objectives : Inhibition of phase I enzymes such as cytochrome P450 (CYP) 1A1 or 1A2 is considered a major mechanism of protection against initiation of carcinogenesis. The inhibition of toxic enzymes and CYP were studied with so many oriental herbral medicine. Recently, numerous polysaccharides and polysaccharide-protein complexes have been isolated from mushrooms and used as a source of therapeutic agents. The most promising biopharmacological activities of these polymers are their immunomodulation and anti cancer. But, in this study the inhibitory effect was on the aqua-acupuncture of Lentinus edodes. Materials : Lentinus edodes aqua-acupuncture solution (LEAS) was prepared and tested for the inhibition of cytochrome P450 (CYP) 1A1 and 1A2 activities. LEAS type I from fruit body of these mushrooms. Type II was extracted from cultured broth of Lentinus edodes mycelum. Results : LEAS type I and type II were significantly inhibited CYP 1A1 and 1A2 enzymes at concentration of 5.0 and 10.0 mg/ml. Conclusion : These results suggested that LEAS may act as block agent against carcinogenesis by inhibition of phase I enzymes.

• 환경생물
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• 제21권1호
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• pp.26-30
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• 2003

해산 어류가 cyochrome P450(CYP)유도제로 알려진 $\beta$-naphthoflavone(BNF)에 의해 어떤 반응을 하는지 살펴보기 위하여, 양식 어류로는 조피볼락(Sebastes schlegeli), 넙치 (Paralichthys olivaceus), 참돔(Pagrus major)을 그리고 자연산 어류로는 숭어(Mugil cephalus)와 쥐치 (Stephanolepis cirrhifer)를 대상으로 조사하였다. 숭어와 쥐치, 참돔의 미크로좀은 DMSO (dimethyl-sulfoxide)로 녹인 BNF를 10 mM이 되도록 첨가하였고, 조피볼락과 넙치의 경우에는 5 mM이 되도록 첨가하여 $30^{\circ}C$의 수조에서 9시간까지 배양하면서 CYP 함량의 경시적인 변화를 조사하였고, 각 어류는 2% 농도로 DMSO 만을 첨가하여 같은 조건으로 배양한 대조구에 대한 상대비(%)로 나타내었다. 그 결과, 대부분의 어류에서는 배양 3~5시간에 최대값을 보이고 이후 줄어들었으나 참돔은 9시간까지도 계속 증가하는 경향을 보였다. 한편,BNF에 의한 CYP 유도 정도는 양식산 어류가 자연산 어류보다 커서 차이를 보였다 즉, 양식산 어류인 넙치, 조피볼락 및 참돔의 CYP 함량은 BNF의 노출농도를 감안하지 않더라도 각 어류의 처음 수준에 비해 207~422%까지 늘었던 것에 비해 자연산 어류인 숭어와 쥐치에서는 206~207%증가하는데 불과하였다. 이처럼 양식산과 자연산 어류에서 CYP 유도제에 의한 CYP 유도정도의 차이는 서식환경의 차이에 따라 생기는 것이라 여겨진다.

• Mass Spectrometry Letters
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• 제8권4호
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• pp.98-104
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• 2017

Ginseng (Panax ginseng Meyer) is a well-known health functional food used as a traditional herbal drug in Asian countries owing to its diverse pharmacological effects. Herb-drug interactions may cause unexpected side effects of co-administered drugs by the alteration of pharmacokinetics through effects on cytochrome P450 activity. In this study, we investigated the herb-drug interactions between Korean red ginseng extract (KRG) and five CYP-specific probes in mice. The pharmacokinetics of KRG extract induced-drug interactions were studied by cassette dosing of five CYP substrates for CYP1A, 2B, 2C, 2D, and 3A and the LC-MS/MS analysis of the blood concentration of metabolites of each of the five probes. The linearity, precision, and accuracy of the quantification method of the five metabolites were successfully confirmed. The plasma concentrations of five metabolites after co-administration of different doses of the KRG extract (0, 0.5, 1, and 2 g/kg) were quantified by LC-MS/MS and dose-dependent pharmacokinetic parameters were determined. The pharmacokinetic parameters of the five metabolites were not significantly altered by the dose of the KRG extract. In conclusion, the single co-administration of KRG extract up to 2 g/kg in vivo did not cause any significant herb-drug interactions linked to the modulation of CYP activity.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9203-9210
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• 2014

Purpose: To explore associations of CYP2E1 and NAT2 polymorphisms with lung cancer susceptibility among Mongolian and Han populations in the Inner Mongolian region. Materials and Methods: CYP2E1 and NAT2 polymorphisms were detected by PCR-RFLP in 930 lung cancer patients and 1000 controls. Results: (1) Disequilibrium of the distribution of NAT2 polymorphism was found in lung cancer patients among Han and Mongolian populations (p=0.031). (2) Lung cancer risk was higher in individuals with c1, D allele of CYP2E1 RsaI/PstI, DraI polymorphisms and slow acetylation of NAT2 (c1 compared with c2, OR=1.382, 95%CI: 1.178-1.587, p=0.003; D compared with C, OR=1.241, 95%CI: 1.053-1.419, P<0.001; slow acetylation compared with rapid acetylation, OR=1.359, 95%CI:1.042-1.768, p=0.056) (3) Compared with c2/c2 and rapid acetylation, c1/c1 together with slow acetylation synergetically increased risk of lung cancer 2.83 fold. (4) Smokers with CYP2E1 c1/c1, DD, and NAT2 slow acetylation have 2.365, 1.916, 1.841 fold lung cancer risk than others with c2/c2, CC and NAT2 rapid acetylation, respectively. (5) Han smokers with NAT2 slow acetylation have 1.974 fold lung cancer risk than others with rapid acetylation. Conclusions: Disequilibrium distribution of NAT2 polymorphism was found in lung cancer patients among Han and Mongolian populations. Besides, Han smokers with NAT2 slow acetylation may have higher lung cancer risk compared with rapid acetylation couterparts. CYP2E1 c1/c1, DD and NAT2 slow acetylation, especially combined with smoking, contributes to the development of lung cancer. CYP2E1 c1/c1 or DD genotype and NAT2 slow acetylation have strong synergistic action in increasing lung cancer risk.