• Molecules and Cells
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• v.43 no.3
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• pp.236-250
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• 2020

Currently, many available anti-cancer therapies are targeting apoptosis. However, many cancer cells have acquired resistance to apoptosis. To overcome this problem, simultaneous induction of other types of programmed cell death in addition to apoptosis of cancer cells might be an attractive strategy. For this purpose, we initially investigated the inhibitory role of TRIP-Br1/XIAP in necroptosis, a regulated form of necrosis, under nutrient/serum starvation. Our data showed that necroptosis was significantly induced in all tested 9 different types of cancer cell lines in response to prolonged serum starvation. Among them, necroptosis was induced at a relatively lower level in MCF-7 breast cancer line that was highly resistant to apoptosis than that in other cancer cell lines. Interestingly, TRIP-Br1 oncogenic protein level was found to be very high in this cell line. Up-regulated TRIP-Br1 suppressed necroptosis by repressing reactive oxygen species generation. Such suppression of necroptosis was greatly enhanced by XIAP, a potent inhibitor of apoptosis. Our data also showed that TRIP-Br1 increased XIAP phosphorylation at serine87, an active form of XIAP. Our mitochondrial fractionation data revealed that TRIP-Br1 protein level was greatly increased in the mitochondria upon serum starvation. It suppressed the export of CypD, a vital regulator in mitochondria-mediated necroptosis, from mitochondria to cytosol. TRIP-Br1 also suppressed shikonin-mediated necroptosis, but not TNF-α-mediated necroptosis, implying possible presence of another signaling pathway in necroptosis. Taken together, our results suggest that TRIP-Br1/XIAP can function as onco-proteins by suppressing necroptosis of cancer cells under nutrient/serum starvation.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.227-233
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• 2021

Benzo[a]pyrene (B[a]P) is a polycyclic aromatic hydrocarbon and ubiquitous environmental toxin with known harmful effects to human health. Abnormal phenotypes of keratinocytes are closely associated with their exposure to B[a]P. Resorcinol is a component of argan oil with reported anticancer activities, but its mechanism of action and potential effect on B[a]P damage to the skin is unknown. In this study, we investigated the effects of resorcinol on B[a]P-induced abnormal keratinocyte biology and its mechanisms of action in human epidermal keratinocyte cell line HaCaT. Resorcinol suppressed aryl hydrocarbon receptor (AhR) activity as evidenced by the inhibition of B[a]P-induced xenobiotic response element (XRE)-reporter activation and cytochrome P450 1A1 (CYP1A1) expression. In addition, resorcinol attenuated B[a]P-induced nuclear translocation of AhR, and production of ROS and pro-inflammatory cytokines. We also found that resorcinol increased nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activity. Antioxidant response element (ARE)-reporter activity and expression of ARE-dependent genes NAD(P)H dehydrogenase [quinone] 1 (NQO1), heme oxygenase-1 (HO-1) were increased by resorcinol. Consistently, resorcinol treatment induced nuclear localization of Nrf2 as seen by Western analysis. Knockdown of Nrf2 attenuated the resorcinol effects on ARE signaling, but knockdown of AhR did not affect resorcinol activation of Nrf2. This suggests that activation of antioxidant activity by resorcinol is not mediated by AhR. These results indicate that resorcinol is protective against effects of B[a]P exposure. The mechanism of action of resorcinol is inhibition of AhR and activation of Nrf2-mediated antioxidant signaling. Our findings suggest that resorcinol may have potential as a protective agent against B[a]P-containing pollutants.

• Environmental Mutagens and Carcinogens
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• v.26 no.3
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• pp.77-85
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• 2006

2,3,7,8-Tetrachlorodibenzo-$\rho$-dioxin(TCDD) displays high toxicity in animals and has been implicated in human carcinogenesis. Although TCDD is recognized as potent carcinogens, relatively little is known about their role in the tumor promotion and carcinogenesis. It is known that TCDD can increase of cancer risk from various types of tissue by a mechanism possibly involving the aryl hydrocarbon receptor (AhR) activation. In this study, effects of TCDD on cellular proliferation of normal human skin and lung fibroblasts, Detroit551 and WI38 cells were investigated. In addition, to enhance our understanding of TCDD-mediated carcinogenesis, we have investigated process in which expression of Erk1/2, cyclinD1, oncogene such as Ha-ras and c-myc, and their cognate signaling pathway. TCDD that are potent activators of AhR-mediated activity was found to induce significant increase of cytochrome P4501A1 mRNA expression, suggesting a presence of functional AhR. These results support that CYP1A1 enzyme may be involved in the generation of TCDD-induced toxicity. Moreover mitogen-activated protein kinases (MARKs) phosphorylation and cyclin D1 overexpression are induced by TCDD, which corresponded with the progression of cellular proliferation. However, TCDD did not affected Ha-ras and c-myc mRNA expression. Taken together, it seems that TCDD are could be a part of cellular proliferation in non-tumorigenic normal human cells such as Detroit551 and WI38 cells through the upregulation of MAPKs signaling pathway regulating growth of cell population. Therefore, AhR-activating TCDD could potentially contribute to tumor promotion and Detroit551 and WI38 cells have been used as a detection system of tumorigenic effects of TCDD.

• Journal of Korean Medicine Rehabilitation
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• v.33 no.3
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• pp.1-15
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• 2023

Objectives We aimed to find out the improvement effect of Baekhogainsam-tang (Baihu Jia Renshen-tang, BIT) on metabolic syndrome and alteration of microbiota and gene expression. Methods We used male C57BI/6 mice and randomly assigned them into three groups. Normal control group was fed 10% kcal% fat diet, high-fat diet (HFD) group was fed 45% kcal% fat diet and 10% fructose water. BIT group was fed same diet as HFD group and treated by BIT for once daily, 6 days per week, total 8 weeks. We measured their body weight and food intake every week and performed oral glucose tolerance test 1 week before the end of the study. Then we collected the blood sample to measure triglyceride, total cholesterol, high-density lipoprotein cholesterol, insulin, and hemoglobin A1c. We harvested tissue of liver, muscle, fat, and large intestine for quantitative polymerase chain reaction (qPCR) and histopathological examination. Fresh fecal samples were collected from each animal to verify alterations of gut microbiota and we used RNA from liver tissue for microarray analysis. Results The body weight and fat weight of BIT group were reduced compared to HFD group. The qPCR markers usually up-regulated in metabolic syndrome were decreased in BIT group. Bacteroides were higher in BIT group than other groups. There were also differences in gene expressions between two groups such as Cyp3a11 and Scd1. Conclusions We could find out BIT can ameliorate metabolic syndrome and suggest its effect is related to gut microbiota composition and gene expression pattern.

• Archives of Pharmacal Research
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• v.28 no.11
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• pp.1287-1292
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• 2005

KR-33028 (N-[4-cyano-benzo[b]thiophene-2-carbonyl]guanidine) is a new cardioprotective agent for preventing ischemia-reperfusion injury. This study was performed to identify the metabolic pathway of KR-33028 in human liver microsomes and to compare its metabolism with that of cryopreserved human hepatocytes. Human liver microsomal incubation of KR-33028 in the presence of NADPH and UDPGA resulted in the formation of four metabolites, M1, M2, M3, and M4. M1 and M2 were identified as 5-hydroxy-KR-33028 and 7-hydroxy-KR-33028, respectively, on the basis of LC/MS/MS analysis with the synthesized authentic standard. M3 and M4 were suggested to be dihydroxy-KR-33028 and hydroxy-KR-33028-glucuronide, respectively. Metabolism of KR-33028 in cryopreserved human hepatocytes resulted in the formation of M1, M2, and M4. These data show a good correlation between major metabolites formed in human liver microsomes and cryopreserved human hepatocytes. In addition, KR­33028 was found to inhibit moderately the metabolism of CYP1A2 substrates. Based on the results obtained metabolic pathway of KR-33028 is proposed.

• Korean Journal of Clinical Pharmacy
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• v.21 no.2
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• pp.90-99
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• 2011

Health Insurance Review & Assessment Service (HIRA) claims database has a high potential to detect signals of new drug interactions. The aim of this study was to evaluate the usefulness of information component (IC) and relative risk (RR) as a tool for signal detection, and to analyze the possible drug interactions caused by clopidogrel using HIRA claims database. This study was performed in elderly patients over 65 years of age who administered clopidogrel from January 2005 to June 2006 in South Korea. Serious Adverse Events (SAEs) as drug interactions of clopidogrel were defined as any ambulatory hospitalization for ischemic diseases within comcomitant medication period of clopidogrel. Information Component (IC) and Relative Risk (RR) were calculated to compare the proportion of drug-SAE pairs in order to select drug specific SAEs. IC and RR signals of clopidogrel drug interaction were screened when IC's 95% confidence interval was greater than 0 and RR's 95% confidence interval was greater than 1 respectively. All detected signals were compared to references such as $Micromedex^{(R)}$ and 2010 Drug Interaction $Facts^{TM}$. Sensitivity, specificity, positive predicted value and negative predicted value were used to evaluate usefulness of this method. Among 13,252,930 cases of elderly patients who co-administered clopidogrel and other drugs, 47,485 cases were detected as SAE. Of these, one-hundred nine cases were detected by the IC-based data-mining approach and ninety one cases were detected by the RR-based data-mining approach. Total One-hundred sixty three unrecognized signals were detected by IC or RR. Twelve signals from IC-based data-mining (57.1%) were corresponded with drug interactions from references and eight signals from RR-based data-mining (38.1%) were corresponded with drug interactions from references. These signals include proton pump inhibitors, calcium channel blockers and HMG CoA reductase Inhibitors, which were known to affect CYP450 metabolism. Further studies using HIRA claims database are necessary to develop appropriate data-mining measure.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.4
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• pp.411-418
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• 2007

Oxidative stress can play a key role in cadmium (Cd)-induced dysfunction. The present study examined the effect of pine needle water extract (PN) on Cd-induced oxidative stress in rats. Sprague-Dawley male rats were divided into three groups: normal group, Cd control group (Cd) and PN-administered Cd group (Cd-PN). $CdCl_2$ in 0.9% NaCl was administered orally with a dose of 5mg/kg of body weight/week, while the PN was administered orally with a dose of 1.26g/kg of body weight/day. Body weight gain was not different between groups, whereas food intakes were significantly lower in the Cd-PN group than in normal or Cd group. Relative liver weight was significantly increased by cadmium administration compared to the normal group. Hepatic cytochrome P450 was significantly lower in Cd and Cd-PN groups than in normal group, while xanthine oxidase and alcohol dehydrogenase activities were significantly higher in the Cd-PN group than in normal or Cd group. Increased hepatic superoxide dismutase, monoamine oxidase, catalase, and glutathione peroxidase activities by cadmium administration were significantly decreased by PN supplement. PN did not affect the hepatic glutathione content in cadmium-administered rats; however, PN significantly lowered the hepatic lipid peroxide level and plasma alanine transferase activity compared to the Cd control group. These results suggest that the PN may alleviate Cd-induced oxidative stress without hepatotoxicity.

• Korean Journal of Ichthyology
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• v.24 no.2
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• pp.118-124
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• 2012

The present study was conducted to investigate different gene expression profile between treated poMSTNpro and non-treated in rainbow trout and to identify those genes that are specifically or predominantly expressed in treated poMSTNpro by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR). We isolated total RNAs in muscle tissues from the treated poMSTNpro fish by immersion bath technique with fish myostatin prodomain (Paralichthys olivaceus, poMSTNpro) for one month and the other was non-treated poMSTNpro, and synthesized cDNA using annealing control primers (ACP, Seegene, Korea). Using 20 different ACPs for PCR, were cloned sequenced, and analyzed identities of 2 differentially expressed genes (DEGs). According to BLAST analysis, sequences of 2 clones significantly matched database entries and confirmed by semi-quantitative RT-PCR. The functional roles of one up-regulated gene, cytochrome P450 mono-oxygenases 2K1v2 (CYP2K1v2), and one down-regulated gene was Profilin-1 were identified. We identified distinctive gene expression profiles in improved rainbow trout growth by treatment with a fish myostatin prodomain using ACP-based GeneFishing.

• The Korean Journal of Pharmacology
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• v.32 no.1
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• pp.75-82
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• 1996

The hypothesis that calcium provoke $O_2^-$ formation by Kupffer cells and may contribute to carbon tetrachloride $(CCl_4)$ induced liver injury was studied in SD rats. In $CCl_4-treated$ animals, hepatic malonaldehyde (nmole/gm liver) and plasma ALT (IU/ml) levels elevated significantly from $119.63{\pm}13.00$ to $268.97{\pm}14.82$ and from $17.3{\pm}0.18$ to $806.08{\pm}37.63$, respectively, compared to those in controls. Activation of Kupffer cells with high dose of retinol (250,000 IU/kg/day, po, for 7 day) significantly enhanced ALT levels, while inactivation of Kupffer cells with gadolinium chloride (7.5 mg/kg/day, ip, for 2 day) attenuated the increase of serum ALT level following $CCl_4$ treatment. Diltiazem (10 mg/kg/day, ip for 2 day) given in combination with retinol led to a marked decrease in ALT levels compare to the level in rats treated only with retinol against $CCl_4$ treatment. In order to determine any alterations in cytochrome P450 activities, the P450 content and the CYP2E1 activity were measured and all $CCl_4-treated$ rats showed significantly lower levels compared to those in controls and vehicle-treated animals. There were significant increases in glutathione peroxidase in all $CCl_4-treated$ rats except diltiazem treated groups. No difference was found among untreated and vehicle-treated rats. It is concluded that Kupffer cells contribute to $CCl_4-induced$ liver injury and that calcium antagonist attenuated the increased $CCl_4-induced$ liver injury due to activation of Kupffer cells.

• Development and Reproduction
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• v.11 no.3
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• pp.195-203
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• 2007

Sex determination and sex differentiation are influenced by genotype in many gonochoristic fish. Cytochrome P450 aromatase (CYP19) is the terminal enzyme in steridogenic pathway that converts androgens into estrogens. In this study, partial fragments of aromatase genes (ovarian aromatase, P450aromA and brain aromatase, P450aromB) were cloned and sequenced in Korean rockfish (Sebastes schlegeli), and gene specific primers were designed based on their sequences. Using these primers, aromatase gene expression during sex differentiation was investigated by RT-PCR. Expression of these aromatase genes were detected both in the head and body parts at 35 dab (days after birth). The number of fish that expressed the aromatase genes decreased at 52 dab, implying down-regulation of these genes. However, these genes were expressed at 59 dab in almost all fish studied here. The expression patterns of both genes are similar throughout the investigated period except for 45 dab where the expression of P450aromB was detected in more fish than that of P450aromA both in the head and body parts. Timing of sex differentiation in this species has been shown to be at around $50{\sim}65$ dab by histological analysis. However, the results from this study suggest that sex differentiation of rockfish may take place $1{\sim}2$ weeks earlier than the period proposed previously. The results also suggest that the mechanism of sex differentiation in viviparous fish may be similar to that in oviparous fish in terms of the importance of aromatase action during the critical period.