• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.727-731
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• 2010

The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD, which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bioindustrial processes.

• The Korean Journal of Physiology and Pharmacology
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• 제21권4호
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• pp.377-384
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• 2017

Activation of protein kinase C (PKC) is closely linked with endothelial dysfunction. However, the effect of $PKC{\beta}II$ on endothelial dysfunction has not been characterized in cultured endothelial cells. Here, using adenoviral $PKC{\beta}II$ gene transfer and pharmacological inhibitors, the role of $PKC{\beta}II$ on endothelial dysfucntion was investigated in cultured endothelial cells. Phorbol 12-myristate 13-acetate (PMA) increased reactive oxygen species (ROS), p66shc phosphorylation, intracellular adhesion molecule-1, and monocyte adhesion, which were inhibited by $PKC{\beta}i$ (10 nM), a selective inhibitor of $PKC{\beta}II$. PMA increased the phosphorylation of CREB and manganese superoxide dismutase (MnSOD), which were also inhibited by $PKC{\beta}i$. Gene silencing of CREB inhibited PMA-induced MnSOD expression, suggesting that CREB plays a key role in MnSOD expression. Gene silencing of $PKC{\beta}II$ inhibited PMA-induced mitochondrial ROS, MnSOD, and ICAM-1 expression. In contrast, overexpression of $PKC{\beta}II$ using adenoviral $PKC{\beta}II$ increased mitochondrial ROS, MnSOD, ICAM-1, and p66shc phosphorylation in cultured endothelial cells. Finally, $PKC{\beta}II$-induced ICAM-1 expression was inhibited by Mito-TEMPO, a mitochondrial ROS scavenger, suggesting the involvement of mitochondrial ROS in PKC-induced vascular inflammation. Taken together, the results suggest that $PKC{\beta}II$ plays an important role in PMA-induced endothelial dysfunction, and that the inhibition of $PKC{\beta}II$-dependent p66shc signaling acts as a therapeutic target for vascular inflammatory diseases.

• Radiation Oncology Journal
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• 제28권2호
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• pp.91-98
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• 2010

목 적: Troglitazone (TRO)은 PPAR-$\gamma$ 작동제로서 온열감수성의 결정에 중요한 요인인 heat shock protein (HSP) 70의 합성을 저해하고 superoxide dismutase (SOD)와 카타라제를 증가시키는 것으로 알려져 있다. 이에 자궁경부암 세포를 대상으로 TRO가 온열감수성에 미치는 영향을 연구하였다. 대상 및 방법: HeLa 세포를 $5{\mu}M$ TRO로 24시간 처치한 후 $42^{\circ}C$에서 1시간 동안 온열처리를 시행하였다. 세포 생존 분획은 clonogenic assay로 측정하였다. 단백질 발현의 변화는 Western blot으로 분석하였다. SOD와 카타라제의 활성도를 측정하였으며, reactive oxygen species (ROS) 는 2',7'-dichlorofluorescin diacetate와 dihydroethidium 를 사용하여 측정하였다. 결 과: 온열처리에 의해 감소된 생존분획이 TRO 전처치에 의해 증가하였다. 온열처리에의해 HSP 70의 발현은 증가하였으나 TRO 전 처치에 의해 감소되지는 않았다. SOD와 카타라제의 활성도가 각각 1.2배, 1.3배 증가하였다. 온열처리에 의해 ROS가 증가하였으며, 증가된 ROS는 TRO 전 처치에 의해 감소하였다. 결 론: TRO는 SOD와 카타라제의 활성도를 증가시키며 이는 온열에 의한 ROS를 감소시켜 결과적으로 온열감수성을 저하시킨다.

• The Korean Journal of Physiology and Pharmacology
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• 제16권6호
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• pp.469-476
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• 2012

2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) is an environmental toxicant with a polyhalogenated aromatic hydrocarbon structure and is one of the most toxic man-made chemicals. Exposure to 2,3,7,8-TCDD induces reproductive toxicity, immunotoxicity, and hepatotoxicity. In this study, we evaluated how 2,3,7,8-TCDD-induced hepatotoxicity affect the expression of heat shock proteins and antioxidant enzymes using the real-time polymerase chain reaction (PCR) in rat. 2,3,7,8-TCDD increased heat shock protein (Hsp27, ${\alpha}$-B-crystallin, Mortalin, Hsp105, and Hsp90s) and antioxidant enzymes (SOD-3, GST and catalase) expression after a 1 day exposure in livers of rats, whereas heat shock protein (${\alpha}$-B-crystallin, Hsp90, and GRP78) and antioxidant enzymes (SOD-1, SOD-3, catalase, GST, and GPXs) expression decreased on day 2 and then slowly recovered back to control levels on day 8. These results suggest that heat shock proteins and antioxidant enzymes were induced as protective mechanisms against 2,3,7,8-TCDD induced hepatotoxicity, and that prolonged exposure depressed their levels, which recovered to control levels due to reduced 2,3,7,8-TCDD induced hepatotoxicity.

• Journal of Veterinary Science
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• 제22권4호
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• pp.54.1-54.13
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• 2021

Background: Hypoxia causes oxidative stress and affects cardiovascular function and the programming of cardiovascular disease. Melatonin promotes antioxidant enzymes such as superoxide dismutase, glutathione reductase, glutathione peroxidase, and catalase. Objectives: This study aims to investigate the correlation between melatonin and hypoxia induction in cardiomyocytes differentiation. Methods: Mouse embryonic stem cells (mESCs) were induced to myocardial differentiation. To demonstrate the influence of melatonin under hypoxia, mESC was pretreated with melatonin and then cultured in hypoxic condition. The cardiac beating ratio of the mESC-derived cardiomyocytes, mRNA and protein expression levels were investigated. Results: Under hypoxic condition, the mRNA expression of cardiac-lineage markers (Brachyury, Tbx20, and cTn1) and melatonin receptor (Mtnr1a) was reduced. The mRNA expression of cTn1 and the beating ratio of mESCs increased when melatonin was treated simultaneously with hypoxia, compared to when only exposed to hypoxia. Hypoxia-inducible factor (HIF)-1α protein decreased with melatonin treatment under hypoxia, and Mtnr1a mRNA expression increased. When the cells were exposed to hypoxia with melatonin treatment, the protein expressions of phospho-extracellular signal-related kinase (p-ERK) and Bcl-2-associated X proteins (Bax) decreased, however, the levels of phospho-protein kinase B (p-Akt), phosphatidylinositol 3-kinase (PI3K), B-cell lymphoma 2 (Bcl-2) proteins, and antioxidant enzymes including Cu/Zn-SOD, Mn-SOD, and catalase were increased. Competitive melatonin receptor antagonist luzindole blocked the melatonin-induced effects. Conclusions: This study demonstrates that hypoxia inhibits cardiomyocytes differentiation and melatonin partially mitigates the adverse effect of hypoxia in myocardial differentiation by regulating apoptosis and oxidative stress through the p-AKT and PI3K pathway.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3015-3023
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• 2012

The mitochondrial antioxidant protein manganese superoxide dismutase (MnSOD) may represent a new type of tumor suppressor protein. Overexpression of the cDNA of this gene by plasmid or recombinant lentiviral transfection in various types of cancer leads to growth suppression both in vitro and in vivo. We previously determined that changes in MnSOD expression had bidirectional effects on adriamycin (ADR) when combined with nitric oxide (NO). Radiation induces free radicals in a manner similar to ADR, so we speculated that MnSOD combined with NO would also have a bidirectional effect on cellular radiosensitivity. To examine this hypothesis, TE-1 human esophageal squamous carcinoma cells were stably transfected using lipofectamine with a pLenti6-DEST plasmid containing human MnSOD cDNA at moderate to high overexpression levels or with no MnSOD insert. Blastidicin-resistant colonies were isolated, grown, and maintained in culture. We found that moderate overexpression of MnSOD decreased growth rates, plating efficiency, and increased apoptosis. However, high overexpression increased growth rates, plating efficiency, and decreased apoptosis. When combined with NO, moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells, whereas high MnSOD overexpression had the opposite effect. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.

• The Korean Journal of Physiology and Pharmacology
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• 제16권6호
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• pp.399-404
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• 2012

We investigated inhibitory effects of extract containing quercetin-3-O-${\beta}$-D-glucuronopyranoside (ECQ) extracted from Rumex Aquaticus Herba on indomethacin-induced gastric damage in Rats. Gastritis was induced in male Sprague-Dawley rats (200~220 g) by oral administration of indomethacin at a dose of 40 mg/kg. One hour before administration of indomethacin, animals were orally pretreated with ECQ at doses of 0.3, 1, 3 or 10 mg/kg. Six hours after indomethacin administration, the rats were sacrificed and the stomach was excised and opened along the greater curvature, and the surface area of gastric lesion was measured using optical microscope. Superoxide dismutase (SOD), catalase (CAT), myeloperoxidase (MPO) activities and malondialdehyde (MDA) levels were measured by ELISA. Western blot analysis was performed to detect protein expression of SOD-2. Linear hemorrhagic mucosal lesions were observed in the stomach 6 hours after oral administration of indomethacin. Pretreatment with ECQ significantly reduced the severity of the lesions in a dose-dependent manner. It also inhibited the reductions in SOD and CAT activities and SOD expression by the indomethacin-induced gastric damage. In addition, the pretreatment with ECQ significantly suppressed the elevation of the MPO activity and the MDA levels induced by indomethacin. These results suggest that ECQ has the inhibitory effects via antioxidative action against indomethacin-induced gastritis in rats.

• 생명과학회지
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• 제16권2호
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• pp.180-185
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• 2006

일품벼(Oryza sativa L. cv. Ilpoombye)에 고선량 감마선을 조사한 후 벼 잎의 생리적 손상과 항산화 효소인 superoxide dismutase (SOD)의 isoenzyme 수준에서의 유전자 발현 및 효소활성 변화와의 연관성을 조사하였다. 500 Gy의 감마선 조사는 24 h 이내에 벼 잎의 단백질, 엽록소, 그리고 카로테노이드의 함량을 유의적으로 감소시켰으며 특히 엽록소는 대조구에 비해 26% 이상 감소하였다. 반면에 SOD isoenzyme들의 유전자 발현은 감마선 조사 후 6 h부터 24 h까지는 전반적으로 대조구보다. 높게 유지되었으나 48 h부터 현저히 감소되어 72h에는 모든 isoenzyme들의 유전자 발현 이 대조구보다. 낮았다. 그러나 isoenzyme들의 효소활성은 조사구에서 일부 CuZn-SOD isoenzyme들의 경우 48 h까지 대조구보다. 약간 높았지만 72h에는 모두 현저히 감소하였다. 따라서 본 연구에 사용된 500 Gy의 고선량 감마선은 단백질, 엽록소, 그리고 카로테노이드 함량의 감소를 초래하며, 조사 후 초기단계에는 이러한 생리적 손상과 무관하게 일시 적으로 SOD isoenzyme들의 유전자 발현을 증가시키지만 72 h 이후에는 유전자 발현과 효소활성을 동시에 감소시키면서 산화스트레스에 의한 생리적 손상을 유도하는 것으로 생각된다.

• BMB Reports
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• 제42권3호
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• pp.136-141
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• 2009

Familial Amyotrophic lateral sclerosis (FALS) is a progressive neurodegenetative disorder induced by mutations of the SOD1 gene. Heat shock protein 27 (HSP27) is well-defined as a stress-inducible protein, however the its role in ALS protection has not yet been established. To investigate the role HSP27 may have in SOD1 mutant-mediated apoptosis, human SOD1 or HSP27 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame fusion protein, which was then transduced into cells. We found the purified PEP-1-HSP27 fusion proteins can be transduced efficiently into neuronal cells and protect against cell death by enhancing mutant SOD1 activity. Moreover, transduced PEP-1-HSP27 efficiently prevents protein aggregation produced by oxidative stress. These results suggest that transduced HSP27 fusion protein may be explored as a potential therapeutic agent for FALS patients.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1996년도 정기총회 및 학술발표회
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• pp.30-30
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• 1996

A suproxide dismutase gene of Aquifex pyroprolus, a novel marine hypenhermophilic bacterium, was cloned, expressed, and characterized. The SOD of A pyrophilus (ApSOD) is an iron-containing homo-oligomeric protein with a monomeric molecular weight of 24.2 kDa. the amino acid sequence is similar to those of known Mn- and Fe-SODs from thermophilic archaea, and metal binding residues in all SOD sequences from different species are also conserved in A. pyrophilus SOD. (omitted)