• 생약학회지
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• 제50권3호
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• pp.159-164
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• 2019

Zanthoxylum Peel is widely used as a common spice for a variety of foods. In the orient, it has also been used as traditional agents for treating diseases such as indigestion. Recently, Zanthoxylum Peel has been reported to have anti-cancer activity, anti-microbial activity, and anti-inflammatory activity. Chemical components are known sanshool compounds and xanthoxylin. In this study, we were carried out to investigate the constituents of inhibiting a drug metabolizing enzyme CYP3A4 from Zanthoxylum Peel. CYP3A4 is known as an enzyme involved in drug metabolism as monooxygenase containing the heme. As a result of experiment, we found that bergapten ($IC_{50}=18.21{\mu}M$) and quercetin ($IC_{50}=17.27{\mu}M$) isolated from EtOAc extract of Zanthoxylum Peel showed remarkable CYP3A4-inhibiting activities. Structures of the isolated active compounds were established by chemical and spectroscopic means.

• Archives of Pharmacal Research
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• 제27권5호
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• pp.547-553
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• 2004

The pyrrolizidine alkaloids (PAs), contained in a number of traditional remedies in Africa and Asia, show wide variations in metabolism between animal species but little work has been done to investigate differences between animal strains. The metabolism of the PA senecionine (SN) in Fischer 344 (F344) rats has been studied in order to compare to that found in the previously investigated Sprague-Dawley (SO) rats (Drug Metab. Dispos. 17: 387, 1989). There was no difference in the formation of ($\pm$) 6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP, bioactivation) by hepatic microsomes from either sex of SO and F344 rats. However, hepatic microsomes from male and female F344 rats had greater activity in the Noxidation (detoxication) of SN by 88% and 180%, respectively, when compared to that of male and female SD rats. Experiments conducted at various pH showed an optimum pH of 8.5, the optimal pH for flavin-containing monooxygenase (FMO), for SN N-oxidation by hepatic microsomes from F344 females. In F344 males, however, a bimodal pattern was obtained with activity peaks at pH 7.6 and 8.5 reflecting the possible involvement of both cytochrome P450 (CYP) and FMO. Use of specific inhibitors (SKF525A, 1-benzylimidazole and methimazole) showed that the N-oxide of SN was primarily produced by FMO in both sexes of F344 rats. In contrast, SN N-oxide formation is known to be catalyzed mainly by CYP2C11 rather than FMO in SD rats. This study, therefore, demonstrated that there were substantial differences in the formation of SN N-oxide by hepatic microsomes from F344 and SD rats and that this detoxification is catalyzed primarily by two different enzymes in the two rat strains. These findings suggest that significant variations in PA biotransformation can exist between different animal strains.

• 생약학회지
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• 제37권2호통권145호
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• pp.97-102
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• 2006

Garlic(Allium sativum Linn) is widely used as a common condiment for a variety of foods and beverages. It has been well known that fresh garlic and garlic supplement of commercial preparations have various therapeutic properties including antimicrobial activity, antiplatelet aggregation, antihypertension, and cholesterol-lowering effects, which contribute to its increasing uses for an alternative medicine. Allicin(diallyl thiosulfinate), the major bioactive components of garlic, is formed by alliinase cleavage of the naturally occurring alliin upon crushing or mincing of garlic, and is the progenitor of a number of other products, such as diallyl disulfide. CYP3A4, heme-containing monooxygenase, is a key enzyme responsible for drug metabolism. Therefor, in the present study, we isolated and examined the compounds with CYP3A4-inhibiting activities from garlic. Among EtOAc extracts of garlic, we found that N-p-coumaroyltyramine and N-feruloyltyramine showed remarkable CYP3A4-inhibiting activities, compared to diallyl disulfide. Structures of the isolated active compounds were established by chemical and spectroscopic means.

• Toxicological Research
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• 제17권
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• pp.127-133
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• 2001

Together with cytochrome P450 (CYP), flavin-containing monooxygenase (FMO) present in liver microsomes oxidizes various endogenous and exogenous chemicals. In an effort to determine the human FMO activity, we have developed two non-invasive urine analysis methods using caffeine (CA) and ranitidine (RA) as the probe compounds. As the production of theobromine (TB) and ranitidine N-oxide (RANO) from CA and RA is catalyzed primarily by the hepatic FMO, we have assigned the urinary molar ratios of TB/CA and RA/RANO as the in vivo FMO activity. In 200 age-matched Korean volunteers, the obtained TB/CA ratio ranged from 0.4 to 15.2 (38-fold difference) and the RA/RANO ratio from 5.7 to 27.2 (4.8-fold). The FMO activity of 20's, determined by caffeine metabolism, was the highest (2.5$\pm$l.9) and those of 30's, 40's, 50's, 60's and 70's were 40%, 50%, 24%, 39% and 36% of the 20's, respectively. Intake of grapefruit juice, known to contain flavonoids, inhibited the in vivo FMO (TB/CA) activity by 79%. Addition of the flavonoids like naringin, quercitrin and kaempferol, present in grapefruit juice, to the in vitro microso-mal FMO assay, thiobenzamide S-oxidation, produced 75%, 70% and 60% inhibition, respectively. Obtained Ki values of quercitrin, kaempferol and naringin on the in vitro FMO activity were 6.2, 12.0 and 13.9 $\mu\textrm{M}$, respectively. This suggested that the dose of drug should need to be adjusted to suit the individual FMO activities when the drugs metabolized by FMO are given to patients. As the intake of grapefruit juice has been identified to inhibit the FMO as well as CYP3A4 and lA2 activities, patients taking drugs metabolized by these enzymes should not drink grapefruit juice as the carrier.

• Journal of Plant Biotechnology
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• 제32권3호
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• pp.167-173
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• 2005

본 연구는 고추열매의 capsaicinoid 생합성 조절 기작을 연구하고자 general phenylpropanoid pathway의 2번째 단계에 작용하는 것으로 알려진 3종류의 c4h cDNA clone을 확보하여 염기서열을 분석한 결과, pc4h1와 pc4h2의 크기는 각각 1,775 bp와 1,655 bp으로 505개의 아미노산으로 구성된 펩티드를 암호하는 full length의 ORF를 갖추고 있었으나 pc4h3는 5'-말단의 coding 영역 일부가 소실 되었다. 이들은 공히 모든 cytochrome P450 효소에서 진화학적으로 보존되어 있는 3종류의 conserved region 즉 domain 1(proline-rich region), domain 2 (threonine-containing binding pocket for the oxygen molecule), 그리고domain 3(heme binding region)을 포함하고 있었으며 evolutionary tree분석을 통하여 pc4h1와 pc4h2는 모두 Class I에 속하는 것으로 서로 간에는 95.8%의 유사성을 나타내어 거의 동일한 유전자인 것으로 조사되었다. 특히 Class II로 분류된 Citrus sinensis C4H1과 Phaseolus vulgaris C4H와는 단지 64.9에서 64.5%의 homology를 나타내었다. 또한 pc4h2는 상처를 주지 않은 조직에서는 비교적 적은 양의 mRNA 발현수준을 보였으나 상처를 가한 후 6시간의 열매에서는 400%, 뿌리에서는 200%까지 그 발현 양이 증가하였다. 이와 유사하게 pc4h1의 전사량도 상처에 의하여 유도는 되나 basal level이 pc4h2보다 높아서 발현증가 비율은 그다지 높지 않았다. 반면에 pc4h3 유전자는 상처를 주지 않은 모든 조직에서 거의 발현되지 않았으며 상처에 의하여서도 전혀 반응을 하지 않았다.

• 한국산업보건학회지
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• 제19권1호
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• pp.73-79
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• 2009

This study was undertaken to investigate the effects of single and combined exposure of toluene (T) and xylene (X) on the cytochrome-450(CYP)-mediated metabolizing capacity, induction of CYP isozymes and the excretion of their metabolites in urine. Animal were adults male Sprague-Dawley (SD) rats and divided into 4 groups such as control, T (treated with 63.7 mg/body kg), X (treated with 65.9 mg/body kg) and TX(T=X). Organic solvents was administrated by intraperitoneal injection for 3 days. The contents of protein and CYP in liver microsomes of control group were $16.48{\pm}0.56 mg/m{\ell}$ and $0.744{\pm}0.025$ nmol/mg protein, respectively, and they contents were significantly lower than in derived from treated groups (p<0.01). The activities of PROD and ${\rho}NPH$ were significantly higher in single treated groups than in control and combined group (TX). When Western immunoblotting were carried out with two monoclonal antibodies (MAb 1-98-1 and MAb 2-66-3) which were specific against CYP2B1/2 and CYP2E1, respectively, a strong signal corresponding to CYP2B1/2 was observed in microsomes obtained from rats treated with X and TX. The color density against CYP2E1 was slightly increased in T and TX groups compared with C and X groups. The amounts of urinary hippuric acid in T single treated group was $3.29{\pm}1.97$ g/g creatinine and TX combined group was $2.91{\pm}1.76$ g/g creatinine, but was not significant. However, amount of urinary methy hippuric acid in X single treated group ($1.62{\pm}0.72$ g/g creatinine) was significantly higher than TX combined group ($0.93{\pm} 0.63$ g/g creatinine)(p<0.01). These results suggested that CYP2E1 isozyme might be responsible for the metabolism of T, and CYP2B1/2 isozyme is for X. And also, difference of metabolites level between single and combined group may be speculated that the intermediates of T and X interacted each other in the process of their metabolite formation reaction.

• 대한약리학회지
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• 제26권2호
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• pp.145-152
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• 1990

최근 본 교실에서는 two-bath system을 이용하여 혈관 내피세포에서 superoxide에 의존한 혈관 이완성 물질을 동정하여 발표하였다. 본 실험에서는 상기 system을 이용하여 돼지의 관상동맥 내피세포에서 유리되는 superoxide에 의존한 이완성 물질이 고양이의 흥부 대동맥 내피 및 소의 대동맥 배양내피세포에서 얻어진 이완성 물질에 의한 이완과 매우 유사함을 관찰하여 다음과 같은 결과를 얻었다. 1. 고양이 흥부 대동맥, 돼지 관상동맥의 내피세포 및 소 대동맥 배양 내피세포 등에서 유리되는 superoxide에 의존한 이완 물질은 모두 유사한 이완 작용을 나타내었다. 2. 돼지 관상 동맥 내피세포에서 유리되는 superoxide 의존성 이완 물질이 고양이의 흥부 대동맥 내피세포나 소의 대동맥 배양 내피세포에서 유리되는 이완 물질과는 다소 다른 점도 있었다. 즉, 돼지 관상동맥 내피세포에서 유리되는 이완 물질의 작용은 catalase나 superoxide dismutase(SOD)에 의하여 억제되었으나, 후자의 두 동맥 내피세포에서 유리되는 이완 물질은 SOD에 의해서만 억제되었다. 3. 이러한 이완성 물질들의 생성은 여러 lipoxygenase억제제인 gossypol, nordihydroguaiaretic acid, AA 861 및 eicosatetraynoic acid 등의 전처치에 의하여 봉쇄되었다. 4. Cyclooxygenase 억제제인 indomethacin이 나 cytochrome P-450 monooxygenase 억제제인 proadifen과 cimetidine에 의하여는 봉쇄되지 아니하였다. 이상의 결과로부터 이러한 이완성 물질들은 비록 각기 다른 종의 동물 모델에서 얻었다고 하더라도 장기에 따라 다소 반응의 차이는 있으나 동질성 이완 물질이며, 나아가 이러한 이완성 물질은 여러 조직의 허혈-재관류 손상에 있어서 병리생리학적으로 관련될 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.83-91
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• 1999

Angiotensin II (ANG II) has a biphasic effect on $Na^+$ transport in proximal tubule: low doses of ANG II increase the $Na^+$ transport, whereas high doses of ANG II inhibit it. However, the mechanisms of high dose ANG II-induced inhibition on $Na^+$ uptake are poorly understood. Thus the aim of the present study was to investigate signal transduction pathways involved in the ANG II-induced inhibition of $Na^+$ uptake in the primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined serum-free medium. ANG II $(10^{-9}\;M)-induced$ inhibition of $Na^+$ uptake was blocked by losartan $(10^{-8}\;M,\;AT_1\;antagonist),$ but not by PD123319 $(10^{-8}\;M,\;AT_2\;antagonist)$ (P<0.05). ANG II-induced inhibition of $Na^+$ uptake was also completely abolished by neomycin $(10^{-4}\;M,$ PLC inhibitor), W-7 $(10^{-4}\;M,$ calmodulin antagonist), and $AACOCF_3\;(10^{-6}\;M,\;PLA_2\;inhibitor)$ (P<0.05). ANG II significantly increased $[^3H]arachidonic$ acid (AA) release compared to control. The ANG II-induced $[^3H]AA$ release was blocked by losartan, $AACOCF_3,$ neomycin, and W-7, but not by PD123319. ANG II-induced $[^3H]AA$ release in the presence of extracellular $Ca^{2+}$ was greater than in $Ca^{2+}-free$ medium, and it was partially blocked by TMB-8 $(10^{-4}\;M,$ intracelluar $Ca^{2+}$ mobilization blocker). However, in the absence of extracellular $Ca^{2+},$ it was completely blocked by TMB-8. In addition, econazole $(10^{-6}\;M,$ cytochrome P-450 monooxygenase inhibitor) and indomethacin $(10^{-6}\;M,$ cyclooxygenase inhibitor) blocked ANG II-induced inhibition of $Na^+$ uptake, but NGDA $(10^{-6}\;M,$ lipoxygenase inhibitor) did not affect it. In conclusion, $PLA_2-mediated$ AA release is involved in ANG II-induced inhibition of $Na^+$ uptake and is modulated by $[Ca^{2+}]_i$ in the PTCs.

• 한국응용곤충학회지
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• 제35권3호
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• pp.254-259
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• 1996

Yumenoshima III 집파리 계통을 ethyl fenitrothion으로 30세대 도태시킨 EF-30 계통에 있어서의 parathion 저항성 메카니즘을 생화학적으로 조사하였다. 아세틸콜린에스테라제 저해활성은 저항성계통과 감수성 SRS 계통간에 커다란 차이를 보여 이 효소의 감수성 저하가 저항성의 주료 메카니즘으로 작용하고 있음을 알 수 있었다. 양 계통에 있어서의 parathion과 paraoxon의 in vitro 분해활성은 미크로좀 및 수용성 분획과 관련이 있으며, 각각 NADPH와 glutathione을 필요로 하였다. 저항성계통은 감수성계통에 비하여 GSH S-transferase 활성이 높아 이 효소가 저항성 메카니즘에 중요한 역할을 하고 있는 것으로 추정되었다. 저항성계통은 parathion에 대하여 101,487배, ethyl parathion에 대하여 25,914배의 저항성비를 나타내어 parathion이 GSH S-transferase의 기질로 작용하고 있음을 알 수 있었다. 이상의 결과로부터 EF-30 계통에 있어서의 저항성 메카니즘에는 수종의 요인이 관여하여 parathion에 대하여 높은 저항성을 나타냄을 알 수 있었으나, 이들 요인이외에 타 요인의 관여를 배제할 수 없었다.

• 생명과학회지
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• 제20권1호
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• pp.133-141
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• 2010

LED의 간 보호 기능을 연구하기 위하여 $CCl_4$ 및 ethanol로 SD rat에 간독성을 유발한 다음, LED를 처리하였다. LED의 간 기능 보호효과는 간장치료제인 Silymarin과 비교하였다. $CCl_4$로 간 독성을 유발한 경우, LED는간의 항산화효소인 SOD, catalase, GSH peroxidase 효소활성의 항진을 유도하였고, 산화물인 TBARS의 함량을 감소시켰다. 또한 간 손상의 지표인 혈장의 GOT, GPT 및 LDH의 활성을 감소시켰다. Ethanol로 간 독성을 유발한 경우 LED는 간의 SOD, catalase, GSH preoxidase 효소활성 및 GSH 함량을 항진시켰고, 총 cholesterol, triglyceride 및 TBARS의 함량을 감소시켰다. 또한 ethanol 대사에 관여하는 ADH 효소 활성을 증진시켰고, ROS 생성에 관여하는 CYP2E1 효소의 발현을 감소시킴으로써, 혈장의 GOT, GPT 및 LDH 효소활성이 감소되었다. 또한 LED는 DPPH 및 mouse liver mitochondrial system에서 항산화효과를 보였다. 이러한 결과로 미루어 볼 때 LED는 in vitro와 in vivo에서 항산화효과에 의한 간 기능 보호효과를 갖는 것으로 추정된다.