• 생명과학회지
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• 제27권6호
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• pp.617-622
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• 2017

본 연구에서는 zebrafish에 사람의 cyt $b_5$ 유전자를 microinjection하여 발현시키고, 그 결과를 형광으로 확인하였다. HeLa cell에서 RT-PCR을 진행한 결과 414 bp의 cyt $b_5$ band가 증폭되었다. 염기서열 분석으로 재확인된 cyt $b_5$ insert를 pEGFP-N3의 형광 vector에 클로닝하였고, 이렇게 준비된 pEGFP-N3-cyt $b_5$ plasmid DNA를 1세 포기의 수정란에 microinjection하였다. cyt $b_5$를 microinjection한 치어를 형광현미경으로 관찰한 결과 대조군 치어보다 훨씬 선명한 형광을 띠는 것이 확인되었다. 최종적으로 치어에서 RNA를 분리하여 RT-PCR하였고 전기영동과 DNA sequencing으로 fusion 단백질의 발현을 재확인하였다. Cyt $b_5$의 발현으로 인해 zebrafish의 생존율이 다소 떨어지는 것으로 확인되었기에 독성 문제를 해결하기 위한 연구가 계속 필요할 것으로 사료된다. 이 연구는 향후 cyt $b_5$가 결핍되었을 경우 발생할 수 있는 여러 질병들을 유전자 차원에서 치료하고, 유용 유전자 클로닝을 위한 기술 개발에 발판이 될 수 있을 것으로 기대된다.

• 생약학회지
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• 제33권1호통권128호
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• pp.35-41
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• 2002

The MeOH ext., $CH_2Cl_2$ Frac., EtOAc Frac., n-BuOH Frac., and $H_2O$ Frac. of 23 Korean medicinal herbs were prepared and were tested the inhibitory effects on Cytochrome P450 (Cyp) 1A1/2, 2B1/2, 2E1. Among the tested samples, the extracts of Selaginella tamariscina, Euonymus alatus, Salvia miltiorhiza, Angelica acutiloba, Rheum palmatum, Paeonia moutan, Scutellaria barbata, Tribulus terrestris, Hedyotis diffusa, Curcuma zedoaria, Rehmania glutinosa, Trogopterus xanthipes, Melandryum firmum, Achyranthes bidentata, Leonurus sibricus, Panax ginseng, Paeonia lactiflora, Poncirus trifoliata, Cnidium officinale, Cyperus rotundus, Corydalis ternata showed significant inhibitory effects on Cyp 1A1/2, 2B1/2, 2E1. The $IC_{50}$ values of those extracts were found to be below $50\;{\mu}g/ml$.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.128-128
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• 1995

To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

• Animal cells and systems
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• 제13권4호
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• pp.453-460
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• 2009

Morphological and molecular classifications were attempted in an effort to establish species-specific classifications of three species of the genus Pholis in Korea; these species were subjected to morphological and molecular methodologies using body measurements, RFLP, RAPD, and phylogenetic trees using the nucleotide sequences of mitochondrial 16S and 12S ribosomal DNAs, cytochrome c oxidase I, and cytochrome b. The data demonstrated that the three species of genus Pholis are distinct from each other, both morphologically and genetically.

• Journal of Ecology and Environment
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• 제43권4호
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• pp.454-461
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• 2019

Background: Ikonnikov's whiskered bat (Myotis ikonnikovi) is found throughout the Korean Peninsula, as well as in Kazakhstan, Russia, Mongolia, China, and Japan. It is small-sized and primarily inhabits old-growth forests. The decrease and fragmentation of habitats due to increased human activity may influence the genetic structure of bat populations. This study was designed to elucidate the population genetic structure of M. ikonnikovi using mitochondrial genes (cytochrome oxidase I and cytochrome b). Results: The results showed that M. ikonnikovi populations from Korea have high genetic diversity. Although genetic differentiation was not detected for the COI gene, strong genetic differentiation of the Cytb gene between Mt. Jeombong and Mt. Jiri populations was observed. Moreover, the results indicated that the gene flow of the maternal lineage may be limited. Conclusions: This study is the first to identify the genetic population structure of M. ikonnikovi. We suggest that conservation of local populations is important for sustaining the genetic diversity of the bat, and comprehensive studies on factors causing habitat fragmentation are required.

• 한국식품영양과학회지
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• 제25권6호
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• pp.969-975
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• 1996

본 연구는 streptozotocin 유발 당뇨쥐의 간과 폐조직에서의 MFO계 활성 변화 및 지질과산화에 미치는 vitamin E의 영향을 관찰하고자 본 실험을 행하였다. 체중 140g 정도의 Sparague-Dawley종 숫컷을 대조군(40mg vitamin E/kg diet group)과 당뇨유발 실험군으로 나눈 후 실험군은 다시 식이내 vitamin E 공급수준에 따라 DM-0E군(0mg vitamin E/kg diet group), DM-40E군(40mg vitamin E/kg diet group), DM-400E군(400mg vitamin E/kg diet group) 나누어 4주간 사육한 후 대조군을 제외한 실험군을 STZ로 당뇨를 유발시켰다. 당뇨유발 6일 후 쥐를 희생시켜 간조직 및 폐조직 microsome 중의 cytochrome P$_{450}$ 및 cytochrome b$_{5}$함량과 NADPH-cytochrome P$_{450}$ reductase 활성도를 측정하고 아울러 microsome내의 지질과산화물을 측정하여 다음과 같은 결과를 얻었다. 1. 간조직 중의 cytochrome P$_{450}$ 함량은 대조군에 비해 DM-0E군, DM-40E 군 및 DM-400E군이 각각 162%, 150%, 56% 증가하였으며 또 DM-400E군은 DM-0E, DM-40E군에 비해 낮았다. 폐조직 중의 cytochrome P$_{450}$ 함량도 간조직에 비해 함량은 낮았지만 경향은 비슷하였다. 2. 간조직 중의 cytochrome b$_{5}$은 대조군에 비해 DM-0E군, DM-40E군은 각각 70%, 53%씩 증가하였으나 DM-400E군은 대조군 수준이었다. 폐조직 중의 cytochrome b$_{5}$함량도 대조군에 비 해 DM-0E, DM-40E 및 DM-400E군에서 각각 149%, 116%, 50%씩 증가하였다. 3. 간조직에서의 cytochrome P$_{450}$ reductase 활성은 대조군에 비해 DM-0E군은 58%, DM-40E군에서는 47% 증가하였으며 DM-400E는 대조군 수준이었다. 폐조직에서도 간조직에서와 비슷한 경향이었다. 4. 간조직의 지질과산화물가는 대조군에 비해 DM-0E군, DM-40E군, DM-400E군이 각각 143%, 95%, 31%씩 높았다. 또 DM-400E군은 DM-0E, DM-40E군에 비해 약 50% 정도 낮았다. 폐조직에서도 경향은 비슷했는데 간조직 보다 과산화적 손상이 다소 높았다. 이상의 결과에서 STZ 유발 당뇨쥐에서는 MFO system의 활성이 증가되고 지질과산화가 촉진되었다. 그러나 식이 중의 vitamin E의 적절한 공급은 MFO system 활성이 저하되고 과산화적 손상이 현저하게 완화됨을 볼 수 있었다.

• Animal cells and systems
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• 제8권3호
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• pp.231-233
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• 2004

Sequences of cytochrome b gene and control region of mitochondrial DNA from Korean common otters (Lutra lutra lutra L.) were examined to provide the genetic information for the conservation of this subspecies. Two haplotypes and one haplotype were revealed in cytochrome b gene and control region, respectively. The available sequences of European common otter (L. l. lutra) from GenBank were compared together with those of Korean common otter in order to determine the degree of sequence variation between them. In cytochrome b gene sequences, two haplotypes from Korea and two haplotypes of Europe showed differences in 12 of 1,045 sites. The Tamura-Nei nucleotide distances between two European haplotypes was 0.10% and those between two Korean haplotypes was also 0.10%, but those between Korean haplotypes and European ones ranged from 0.96% to 1.16%. In the control region, one Korean haplotype and seven European ones showed differences in seven of 300 sites; the Tamura-Nei distances among seven European haplotypes were 0.34% to 1.01%, but those between Korean haplotype and European ones ranged from 1.01% to 1.69%. Although further molecular and morphological studies with specimens from eastern Asia including Amur region and northeast China are needed, it is possible that the Korean common otter might be closer or identical to the far-eastern Asian common otter, L. l. amurensis Dybowski.

• BMB Reports
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• 제34권2호
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• pp.114-117
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• 2001

Cervi Parvum Cornu is widely used as a hemopoietic, tonifying, growth-promoting, cardiotonic, and immuno-modulating agent in Korea. In order to develop the quality control method of Cervi Parvum Cornu by the identification of the biological source or origin, the molecular approach was applied using PCR (polymerase chain reaction) and PCR-RFLF (PCR-restriction fragment length polymorphism) analysis. In the PCR analysis of the mitochondrial 12S rRNA gene and cytochrome b gene regions, no distinctive DNA bands from Cervidae (deer) antlers and Rangifer (reindeer) antlers were observed. However, when the amplified products in the mitochondrial cytochrome b gene region were subjected to restriction digestion with TaqI, Cervidae antlers showed an undigested state of 380 by band, differently from two bands of 230 by and 1S0 by from Rangifer antlers. Based on this finding, the base sequences of amplified PCR products in the range of mitochondria) cytochrome b gene from Cervidae antlers and Rangifer antlers were determined and subjected to restriction analysis by various endonucleases. The results showed that antlers from Rangifer species could be simply discriminated with other antlers from 8 Cervidae species (Chinese deer, Russian deer, Hong Kong deer, New Zealand deer, Kazakhstan deer, elk, red deer and Sika deer) by PCR-RFLP analysis using AtuI, HaeIII, HpaII or Sau3AI(MboI) as well as TaqI in the range of the mitochondrial cytochrome b gene.

• Archives of Pharmacal Research
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• 제27권2호
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• pp.199-205
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• 2004

Cytochrome P450 (P450) 1 enzymes such as P450 1A1, 1A2, and 181 are known to be involved in the oxidative metabolism of various procarcinogens and are regarded as important target enzymes for cancer chemoprevention. Previously, several hydroxystilbene compounds were reported to inhibit P450 1 enzymes and were rated as candidate chemopreventive agents. In this study, we investigated the inhibitory effect of 2-[2-(3,5-dimethoxyphenyl)vinyl]-thiophene (DMPVT), produced from the chemical modification of oxyresveratrol, on the activities of P450 1 enzymes. The inhibitory potential by DMPVT on the P450 1 enzyme activity was evaluated with the Escherichia coli membranes of the recombinant human cytochrome P450 1A1, 1A2, or 1B1 coexpressed with human NADPH-P450 reductase. DMPVT significantly inhibited ethoxyresorufin O-deethylation (EROD) activities with $IC_{50}$ values of 61, 11, and 2 nM for 1A1, 1A2, and 1B1, respectively. The EROO activity in OMBA-treated rat lung microsomes was also significantly inhibited by OMPVT in a dose-dependent manner. The modes of inhibition by DMPVT were non-competitive for all three P450 enzymes. The inhibition of P450 1B1-mediated EROD activity by OMPVT did not show the irreversible mechanism-based effect. The loss of EROD activity in P450 1B1 with OMPVT incubation was not blocked by treatment with the trapping agents such as glutathione, N-acetylcysteine, or dithiothreitol. Taken together, the results suggested DMPVT to be a strong noncompetitive inhibitor of human P450 1 enzymes that should be considered as a good candidate for a cancer chemopreventive agent in humans.

• 약학회지
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• 제23권2호
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• pp.111-118
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• 1979

It was previously reported that cytochrome P$_{450}$ content in liver was increased when Capsicum acetone extract was given chronically to rats. The present study is aimed to investigate the effect of capsaicin, a principal component of red pepper, on the drug metabolizing enzymes in rat liver. Capsaicin (5mg/kg) was given intraperitoneally once a day for seven days and zoxazolamine paralysis time and hexobarbital sleeping time were determined 24 hrs after the last dose of capsaicin. Plasma hexobarbital concentration was also determined five and 15 min after hexobarbital administration to rats. Zoxazolamine paralysis time and hexobarbital sleeping time were shortened by 31.6% and 37.1%, respectively, compared with control group. Plasma hexobarbital concentration was lowered by 26.2% after five min and by 35.2% after 15 min, respectively, compared with control group. However, administration of single dose of capsaicin did not affect the zoxazolamine paralysis time and hexobarbital sleeping time. Microsomal cytochrome P$_{450}$ content and NADPH-cytochrome C reductase activity were increased by 14.6% and 11.6%, respectively in the rats pretreated with capsaicin for seven days, while cytochrome b$_{5}$ content was not changed. These results suggest that treatment with capsaicin for seven days may induce the drug metabolizing enzyme in rat liver.