• KSBB Journal
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• 제11권4호
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• pp.453-461
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• 1996

간세포를 in vitro에서 효율적으로 장기간 배양 및 증식을 유도하여 이용하는 기술을 수립하기 위해서 본 연구에서는 일차배양한 쥐 간세포에 중요한 세포외 기질(ECM)인 collagen이 간세포의 장기배양시 생존능과 기능유지에 미치는 영향을 분석하고자 하였다. 간세포를 in vitrovo에서 40여일 간의 장기간 배양시에 일반적으로 사용되고 있는 단층 배양(single gel)법과 in vivo 상태의 세포 극성에 유사한 환경을 제공하는 복층 배양(sandwich gel)법으로 배양한 경우에 미치는 영향을 서로 비교하여 보았다. 이를 위해서 간세포의 생존 상태를 비교하였요며 나아가 간세포의 특성인 알부민, 파이브로넥틴, 피브리노겐 등의 단백질 생산 빛 분비기능 측정 그리고 요소 생산능과 암모니아 제거능 등을 비교하였다. 복층 배양법이 단층 배양법보다 알부민, 파이브로넥틴, 피브 리노겐 등의 분비기능을 훨씬 효율적으로 유지시키는 것을 관찰할 수 있었으며 또한 요소 생산능과 암모니아 제거능도 훨씬 더 효율적으로 장기간 유지시 킬 수 있음을 확인할 수 있었다. 이로써 in vitro에서 간세포의 기능을 효율척으로 유지시키기 위해서 간세포 배양시 3차원적 공간을 유지시키는 것이 중 요한 요인 중의 하나임을 확인할 수 있었다.

• 한국식품저장유통학회지
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• 제30권3호
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• pp.538-545
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• 2023

본 연구는 probiotics mixture(Lactobacillus acidophilus PBS066(DSM 24936), Lactiplantibacillus plantarum PBS067(DSM 24937), Limosilactobacillus reuteri PBS072(DSM 25175))의 안전성 및 탄수화물 소화 효소 저해 효과를 in vitro로 평가하는 것을 목적으로 한다. 모든 균주는 유럽식품안전처(EFSA) 지침에서 권장하는 항생제 내성 프로필을 충족시켰으며, 모든 균주는 용혈 활성 및 세포 독성을 나타내지 않았다. Probiotics mixture, L. plantarum PBS067 그리고 L. reuteri PBS072는 α-amylase 활성을 억제하는 것으로 나타났으며, probiotics mixture와 이의 균주 3종 모두 α-glucosidase 활성을 억제하는 것으로 나타났다. 이로써 본 연구에 사용한 probiotics mixture 및 이의 단일 균주 3종의 안전성과 탄수화물 소화 억제 효과를 확인하였으며, 따라서 probiotics mixture 이의 단일 균주 3종은 섭취하여도 안전하며 섭취 시 체내 탄수화물 대사를 잠재적으로 조절하는 것을 확인하였다.

• Archives of Pharmacal Research
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• 제28권12호
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• pp.1365-1375
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• 2005

The antiestrogenic effects of marijuana smoke condensate (MSC) and three major cannabinoids, i.e., $\bigtriangleup^{9}$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN), were evaluated using in vitro bioassays, viz., the human breast cancer cell proliferation assay, the recombinant human estrogen receptor (ER) competitive binding assay, and the reporter gene assay. The inhibitory effects on estrogen were also examined using the ethoxyresorufin-O­deethylase (EROD) assay, the aromatase assay, and the 17$\beta$-estradiol ($E_{2}$) metabolism assay. The results showed that MSC induced the antiestrogenic effect via the ER-mediated pathway, while THC, CBD, and CBN did not have any antiestrogenic activity. This suggests that the combined effects of the marijuana smoke components are responsible for the antiestrogenicity of marijuana use. In addition, MSC induced the CYP1A activity and the $E_{2}$ metabolism, but inhibited the aromatase activity, suggesting that the antiestrogenic activity of MSC is also related to the indirect ER-dependent pathway, as a result of the depletion of the in situ $E_{2}$ level available to bind to the ER. In conclusion, pyrogenic products including polycyclic aromatic hydrocarbons (PAHs) in the non-polar fraction, which is the most biologically active fraction among the seven fractions of MSC, might be responsible for the antiestrogenic effect.

• 대한의생명과학회지
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• 제5권1호
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• pp.67-74
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• 1999

주ㆍ야 (day, night phase) 시차에 따른 toluene 대사를 검토할 목적으로 흰쥐에 50% toluene (olive oil과 toluene의 동량혼합액)을 체중 100 g 당 0.2 ml씩 밤 12시, 낮 12시에 각각 투여한 다음 각각 8시간 후에 처치하여, 다음과 같은 결과를 얻었다. 간 microsomal aniline hydroxylase 활성은 night phase군이 day phase군 보다 대조군 및 toluene 투여군 모두 높게 나타났으며, cytosol의 benzylalcohol dehydrogenase (BADH) 활성은 night phase군과 day phase군간에 별다른 차이를 볼 수 없었으나, toluene 투여시에는 night phase군이 day phase군 보다 효소의 활성도가 오히려 억제되었다. 또한 간 조직의 benzaldehyde dehydrogenase (BALDH) 활성은 night phase군이 day phase군 보다 다소 높게 나타나는 경향을 보였으며, toluene 투여시에는 night phase군에서는 대조군 보다 오히려 감소되었으나 day phase 군에서는 높게 나타나는 경향을 보였다. 그리고 in vitro 시험에서 benzylalcohol 및 benzaldehyde가 BADH 및 BALDH 활성을 억제시킴이 관찰되었다. 요 중 hippuric acid 함량은 대조군 및 toluene 투여군 모두 night phase군과 day phase군간에는 별다른 차이를 볼 수가 없었다. 한편 toluene 투여로 인한 체중 당간 무게 및 혈청 xanthine oxidase활성 증가율은 night phase군이 day phase군 보다 높게 나타났다. 이상 실험결과를 종합해 볼 때 요 중 hippuric acid함량이 주ㆍ야 시차에 따라 별다른 차이가 없는데도 불구하고 toluene으로부터 benzylalcohol 및 benzaldehyde 생성률이 day phase군 보다 night phase군이 높게 나타남으로서 toluene 대사산물에 의한 간 손상 정도가 day phase군 보다 night phase군이 높게 나타난 것으로 사료되어 진다.

• 약학회지
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• 제43권6호
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• pp.834-842
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• 1999

The metabolism and pharmacokinetics of M-l, which is metabolite of pentoxifylline, have been studied in human liver microsomes. Biphasic kinetics was observed from the Eadie-Hofstee plot for the formation of both metabolites of M-l. For the kinetics of pentoxifylline, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 1,648 and 5,622 nmol/min/mg protein, and the estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.180 and 4.829 mM, respectively. For M-3, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 0.062 and 0.491 nmol/min/mg protein, and estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.025 and 1.216 mM. The formations of pentoxifylline and M-3 from M-1 were indentified by using several selective inhibitors of cytochrome P450 isoformes at 0.05-5 mM concentration of M-1 in human liver microsomes. For the analysis of low (0.05 mM) concentration of M-1, where the affinity was expected as low, indicated that CYPlA2 and CYP3A4 were major P450 isoforms responsible for pentoxifylline and M-3 formation. CYP3A4 and CYP2A6 appeared to be P450 isoforms responsible for M-3 formation at high (5 mM) concentration of M-1.

• Archives of Pharmacal Research
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• 제16권2호
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• pp.89-93
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• 1993

Incubation of $S(-)-^3H$-nicotine with rabbit lung microsomes in the presence of dioxygen and NADPH results in the formation of metabolities that bind covalently to microsomal macro-molecules. The addition of cytochrome P-450 monooxygenase inhibitors, $\alpha$-methylbenzyl ami-nobenzotriazole and aroclor 1260, inhibited both (S)-nicotine metabolism and covalent binding. The relative rates of oxidation of nicotine $\Delta^{1',5'}$ iminium ion to continine indicates that lung $100,000\times{g}$ supematant catalyzed this oxidation approximately 18 times slower than that of liver system based on mg of protein, and increased covalent interactions. Since than that of liver system based on mg of protein, nd increased covalent interactions. Since the activity of lung iminium oxidase appears much lowr than the liver, it is tempting to speculate that localized concentrations of nicotine $\Delta^{1',5'}$ iminium ion in the lung will survive for a longer period of time. These results support that cytochrome P-450 catalyzed oxidation of nicotine leads to the formation of reactive nad electrophilic intemediates capable of chemical interactions with biomacromolecules.

• 농약과학회지
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• 제3권2호
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• pp.37-46
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• 1999

침투성 살충제 carbofuran의 작물체중 잔류성과 그 안전성을 평가하기 위하여 carbofuran을 살포했을 것으로 간주되는 감자와 땅콩 및 마늘 시료를 주산지 부근의 대규모 시장에서 수집한 후 carbofuran과 carbofuran의 식물체중의 주 대사산물로 보고된 3-hydroxycarbofuran의 잔류량을 조사하고 carbofuran의 각 작물체내 대사 양상을 구명하기 위하여 각 작물의 phesphate buffer 추출액중 carbofuran의 분해실험을 수행하였다. 여러 지역의 시장으로부터 수집한 총 20 점의 완숙마늘 시료중 2점 (M-12와 M-16)의 시료에서 0.13과 0.07mg/kg의 carbofuran이 검출되어 10%의 검출빈도를 보였으나, 그 잔류량은 잔류허용기준인 0.5 mg/kg 보다 훨씬 작았다. 3-Hydroxycarbofuran은 총 20점의 시료중 1점 (M-12)의 시료에서 0.13 mg/kg이 검출되어 5%의 검출빈도를 보였으며, 풋마늘과 감자 및 땅콩 시료중 carbofuran과 3-hyoxycarbofuran의 잔류량은 분석결과 모든 시료에서 검출한계 미만이었다. 완숙마늘의 1일 섭취량에 근거한 carbofuran의 최대 섭취추정량은 0.0013mg으로 1일 최대섭취허용량인 0.55 mg의 0.24%이었으며, 마늘과 감자 및 땅콩시료에서 carbofuran이 검출되지 않아 마늘과 감자 및 땅콩 경작시 carbofuran을 사용하여도 생산물은 안전한 것으로 평가되었다. Carbofuran은 각 작물의 phosphate buffer 추출액중에서 주로 가수분해에 의하여 주 대사산물인 carbofuran phenol (m/z 164)을 생성하였으며, 그 양은 배양기간에 비례하여 증가하였다.

• Toxicological Research
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• 제6권1호
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• pp.51-59
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• 1990

Effects of altering hepatic mixed-function oxidase (MFO) enzyme activities on the metabolism and acute toxicity of parathio were investigated in adult female rats. In vitro hepatic metabolism of parathion to paraoxon was increased by phenobarbital pretreatment (50 mg/kg/day, ip, for 4 consecutive days) and SKF 525-A (50 mg/kg, ip, 1 hr prior to sacrifice) decreased paraoxon formation indicating that phenobarbital induces that form(s) of cytochrome P-450 catalyzing conversion of parathion to paraoxon. Degradation of paraoxon to p-nitrophenol was increased by phenobarbital pretreatment, but not affected by SKF 525-A suggesting that MFO activities play only a minor role in the detoxification of the active metabolite of this insecticide. The phenobarbital-induced increase in paraoxon formation was partially antagonized by SKF 525-A. Significant activity for both parathion activation and paraoxon degradation was also observed in the lung preparation, however, this extrahepatic parathion and paraoxon metabolizing activity was not induced by phenobarbital or inhibited by SKF 525-A pretreatment. Phenobarbital pretreatment increased paraoxon level in livers of rats when measured 3 hr following parathion injection (2 mg/kg, ip). SKF 525-A did not alter parathion or paraoxon levels in brain, blood and liver. Phenobarbital pretreatment decreased the toxicity of parathion (4mg/kg, ip) or paraoxon (1.5 mg/kg, ip) as determined by decreases in lethality and inhibition of brain and lung acetylcholinesterases. An additional SKF 525-A treatment failed to decrease the protective effects of phenobarbital against parathion or paraoxon toxicity. These results suggest that some unknown factors other than hepatic MFO induction are involved in the protective action of phenobarbital against parathion and paraoxon toxicity.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.919-925
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• 2003

Alcohol consumption has numerous health consequences for the human body. For example, heavy drinking on a daily basis causes liver diseases, and certain products such as acetaldehyde produced from alcohol metabolism are more toxic than alcohol itself. Accordingly, the current study evaluated the role of Lactobacillus fermentum MG590 to enhance the removal of the toxic effect of alcohol in alcohol metabolism. The maximum activities of the alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) by L. fermentum MG590 were observed after 6 h of culture. The production of ADH and ALDH by L. fermentum MG590 was also confirmed by SDS-PAGE. Six hours after the addition of alcohol to a culture broth of L. fermentum MG590, the alcohol concentration decreased from 7.5 to 2.7％. From an in vitro evaluation based on hepatocytes, the viability of hepatocytes in a medium containing alcohol and the cytosol of L. fermentum MG590 was higher than that in a medium containing only alcohol. From an in vivo test using SD rats fed a 22％ alcoholic drink, the blood alcohol concentration (BAC), glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) in the rats fed a medium containing L. fermentum MG590 were lower than those in the rats fed a medium containing only the alcohol drink. These results demonstrate that the ADH and ALDH produced by L. fermentum MG590 play an important role in detoxicating alcohol in vivo. Therefore, a fermentation broth of L. fermentum MG590 could be used as an effective alcohol detoxification drink.

• Biomolecules & Therapeutics
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• 제32권1호
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• pp.94-103
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• 2024

Non-alcoholic fatty liver disease (NAFLD) is characterized by excessive accumulation of fat in the liver, and there is a global increase in its incidence owing to changes in lifestyle and diet. Recent findings suggest that p53 is involved in the development of non-alcoholic fatty liver disease; however, the association between p53 expression and the disease remains unclear. Doxorubicin, an anticancer agent, increases the expression of p53. Therefore, this study aimed to investigate the role of doxorubicin-induced p53 upregulation in free fatty acid (FFA)-induced intracellular lipid accumulation. HepG2 cells were pretreated with 0.5 ㎍/mL of doxorubicin for 12 h, followed by treatment with FFA (0.5 mM) for 24 h to induce steatosis. Doxorubicin pretreatment upregulated p53 expression and downregulated the expression of endoplasmic reticulum stress- and lipid synthesis-associated genes in the FFA -treated HepG2 cells. Additionally, doxorubicin treatment upregulated the expression of AMP-activated protein kinase, a key modulator of lipid metabolism. Notably, siRNA-targeted p53 knockdown reversed the effects of doxorubicin in HepG2 cells. Moreover, doxorubicin treatment suppressed FFA -induced lipid accumulation in HepG2 spheroids. Conclusively, these results suggest that doxorubicin possesses potential application for the regulation of lipid metabolism by enhance the expression of p53 an in vitro NAFLD model.