• Animal Bioscience
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• 제34권3_spc호
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• pp.463-470
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• 2021

Objective: This experiment was conducted to find out the immunological effects of wheat phytase when long-chain inorganic polyphosphate (polyP) treated with wheat phytase was added to a macrophage cell line, Raw 264.7, when compared to intact long-chain polyP. Methods: Nitric oxide (NO) production of Raw 264.7 cells exposed to P700, a long-chain polyP with an average of 1,150 phosphate residues, treated with or without wheat phytase, was measured by Griess method. Phagocytosis assay of P700 treated with or without phytase in Raw 264.7 cells was investigated using neutral red uptake. The secretion of tumor necrosis factor α (TNF-α) by Raw 264.7 cells with wheat phytase-treated P700 compared to intact P700 was observed by using Mouse TNF-α enzyme-linked immunosorbent assay kit. Results: P700 treated with wheat phytase effectively increased NO production of Raw 264.7 cells by 172% when compared with intact P700 at 12 h exposure. At 5 mM of P700 concentration, wheat phytase promoted NO production of macrophages most strongly. P700, treated with wheat phytase, stimulated phagocytosis in macrophages at 12 h exposure by about 1.7-fold compared to intact P700. In addition, P700 treated with wheat phytase effectively increased in vitro phagocytic activity of Raw 264.7 cells at a concentration above 5 mM when compared to intact P700. P700 dephosphorylated by wheat phytase increased the release of TNF-α from Raw 264.7 cells by 143% over that from intact P700 after 6 h exposure. At the concentration of 50 μM P700, wheat phytase increased the secretion of cytokine, TNF-α, by 124% over that from intact P700. Conclusion: In animal husbandry, wheat phytase can mitigate the long-chain polyP causing damage by improving the immune capabilities of macrophages in the host. Thus, wheat phytase has potential as an immunological modulator and future feed additive for regulating immune responses caused by inflammation induced by long-chain polyP from bacterial infection.

• 대한본초학회지
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• 제36권6호
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• pp.17-25
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• 2021

Objectives : Baicalein (3,3', 4', 5, 6-pentahydroxyflavone), a type of flavonoid, is a well-known antioxidant and anti-inflammatory ingredient found in Scutellaria baicalensis root. The aim of this study is to investigate the effect of baicalein on nitric oxide (NO) production in TM3 mouse Leydig cells stimulated by indomethacin (IN). Methods : TM3 cells were treated with IN (0.5 μM) and baicalein at concentrations of 12.5, 25, 50, and 100 μM for 24 hr, 40 hr, 42 hr, 44 hr, and 64 hr. After treatments, cell viabilities were measured with the modified MTT assay. The production of nitric oxide in cells was measured by Griess reagent assay. Results : Baicalein showed no cytotoxicity on IN-stimulated TM3. NO production in IN-stimulated TM3 treated for 24 hr with baicalein at concentrations of 12.5, 25, 50, and 100 μM was 95.8%, 94.86%, 89.97%, and 81.52% of the control group treated with IN only, respectively; NO production for 40 hr was 97.34%, 97.34%, 95.15%, and 87.42%, respectively; NO production for 42 hr was 89.12%, 90.14%, 89.74%, and 90.26%, respectively; NO production for 44 hr was 83.83%, 84.94%, 85.65%, and 86.85%, respectively; NO production for 64 hr was 94.12%, 95.38%, 94.21%, and 94.12%, respectively. Specifically, baicalein at concentrations of 12.5, 25, and 50 have been shown to most efficiently inhibit NO productions in 48 hr of treatment. Conclusions : Baicalein might have anti-toxicant effect on Leydig cells related with its inhibition of NO production in Leydig cells stimulated with IN.

• Journal of Periodontal and Implant Science
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• 제53권1호
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• pp.54-68
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• 2023

Purpose: The purpose of this study was to determine whether metformin (MF) could alleviate the expresssion of reactive oxygen species (ROS) and improve the osteogenic ability of bone marrow mesenchymal stem cells derived from diabetic rats (drBMSCs) in vitro, and to evaluate the effect of MF on the ectopic osteogenesis of drBMSCs in a nude mouse model in vivo. Methods: BMSCs were extracted from normal and diabetic rats. In vitro, a cell viability assay (Cell Counting Kit-8), tests of alkaline phosphatase (ALP) activity, and western blot analysis were first used to determine the cell proliferation and osteogenic differentiation of drBMSCs that were subjected to treatment with different concentrations of MF (0, 50, 100, 200, 500 µM). The cells were then divided into 5 groups: (1) normal rat BMSCs (the BMSCs derived from normal rats group), (2) the drBMSCs group, (3) the drBMSCs + Mito-TEMPO (10 µM, ROS scavenger) group, (4) the drBMSCs + MF (200 µM) group, and (5) the drBMSCs + MF (200 µM) + H₂O₂ (50 µM, ROS activator) group. Intracellular ROS detection, a senescence-associated β-galactosidase assay, ALP staining, alizarin red staining, western blotting, and immunofluorescence assays were performed to determine the effects of MF on oxidative stress and osteogenic differentiation in drBMSCs. In vivo, the effect of MF on the ectopic osteogenesis of drBMSCs was evaluated in a nude mouse model. Results: MF effectively reduced ROS levels in drBMSCs. The cell proliferation, ALP activity, mineral deposition, and osteogenic-related protein expression of drBMSCs were demonstrably higher in the MF-treated group than in the non-MF-treated group. H₂O₂ inhibited the effects of MF. In addition, ectopic osteogenesis was significantly increased in drBMSCs treated with MF. Conclusions: MF promoted the proliferation and osteogenic differentiation of drBMSCs by inhibiting the oxidative stress induced by diabetes and enhenced the ectopic bone formation of drBMSCs in nude mice.

• 대한본초학회지
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• 제38권4호
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• pp.11-19
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• 2023

Objectives : The aim of this study was to investigate the effect of baicalein (BA) on the production of hydrogen peroxide and nitric oxide (NO) in RAW 264.7 mouse macrophages stimulated with polyinosinic-polycytidylic acid (poly-IC) and lipoteichoic acid. Methods : RAW 264.7 co-stimulated with poly-IC and lipoteichoic acid were incubated with baicalein at concentrations of 25 and 50 μM. Incubation time is 16 h, 18 h, 20 h, 22 h, and 24 h. After incubation, The production of hydrogen peroxide in RAW 264.7 was measured with dihydrorhodamine 123 assay. Chrysin was used as a comparative material. NO production was evaluated by griess assay. Results : For 16 h, 18 h, 20 h, 22 h, and 24 h incubation, BA at the concentration of 25 and 50 μM significantly inhibited the production of hydrogen peroxide in RAW 264.7 stimulated by poly-IC and lipoteichoic acid (p <0.001). In details, production of hydrogen peroxide in 'poly-IC and lipoteichoic acid'-stimulated RAW 264.7 treated for 16 h with BA at concentrations of 25 and 50 μM was 82.36% and 77.24% of the control group treated with poly-IC and lipoteichoic acid only, respectively; the production of hydrogen peroxide for 18 h was 83.15% and 77.91%, respectively;production of hydrogen peroxide for 20 h was 82.88% and 77.82%, respectively; production of hydrogen peroxide for 22 h was 83.27% and 78.17%, respectively; production of hydrogen peroxide for 24 h was 83.54% and 78.35%, respectively. Additionally, BA at the concentration of 50 and 100 μM significantly inhibited NO production in lipoteichoic acid-induced RAW 264.7 (p <0.001). Conclusions : BA might have anti-oxidative activity related to its inhibition of hydrogen peroxide production in 'poly-IC and lipoteichoic acid'-stimulated RAW 264.7 macrophages.

• 대한본초학회지
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• 제38권6호
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• pp.45-52
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• 2023

Objectives : Effects of baicalein (BA) on oxidative stress in RAW 264.7 mouse macrophages stimulated with peptidoglycan (PG) and lipopolysaccharide (LPS) were investigated. Methods : RAW 264.7 co-stimulated with LPS and PG were incubated with BA at concentrations of 25 and 50 µM. Incubation time was 18 h, 20 h, 22 h, 24 h, and 26 h. After incubation, the production of hydrogen peroxide in RAW 264.7 was measured with dihydrorhodamine 123 assay. Additionally, RAW 264.7 stimulated with PG were incubated with BA at concentrations of 25 and 50 µM for 24 h. After incubation, NO production was evaluated by griess reagent assay. Results : BA significantly inhibited hydrogen peroxide productions (p <0.05). In details, production of hydrogen peroxide in 'LPS and PG'-stimulated RAW 264.7 treated for 18 h with BA at concentrations of 25 and 50 µM was 91.27% and 89.22% of the control group treated with LPS and PG only, respectively; the production of hydrogen peroxide for 20 h was 92.19% and 90.58%, respectively; production of hydrogen peroxide for 22 h was 91.69% and 89.89%, respectively; production of hydrogen peroxide for 24 h was 92.4% and 90.19%, respectively; production of hydrogen peroxide for 26 h was 91.7% and 89.04%, respectively. Additionally, BA at the concentration of 50 and 100 µM significantly inhibited NO production in PG-induced RAW 264.7 (p <0.05). Conclusions : BA might have anti-oxidative activity related to its inhibition of hydrogen peroxide production in 'LPS and PG'-stimulated RAW 264.7 macrophages.

• 대한본초학회지
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• 제39권4호
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• pp.37-45
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• 2024

Objectives : This study aimed to elucidate antioxidant activity of chrysin in polyinosinic-polycytidylic acid (poly-IC) and lipoteichoic acid-induced RAW 264.7 mouse macrophages. Methods : RAW 264.7 co-stimulated with poly-IC and lipoteichoic acid were incubated with chrysin at concentrations of 25 and 50 µM. Hydrogen peroxide production was measured with dihydrorhodamine 123 assay. Nitric Oxide (NO) production was evaluated by griess reagent assay. Results : For 16 h, 18 h, 20 h, 22 h, and 24 h incubation, chrysin at the concentration of 25 and 50 µM significantly suppressed hydrogen peroxide production in poly-IC and lipoteichoic acid-induced RAW 264.7. In details, production of hydrogen peroxide in 'poly-IC and lipoteichoic acid'-stimulated RAW 264.7 treated for 16 h with chrysin at concentrations of 25 and 50 µM was 83.84% and 79.3% of the control group treated with poly-IC and lipoteichoic acid only, respectively; the production of hydrogen peroxide for 18 h was 84.36% and 79.93%, respectively; production of hydrogen peroxide for 20 h was 85.68% and 80.22%, respectively; production of hydrogen peroxide for 22 h was 85.81% and 79.95%, respectively; production of hydrogen peroxide for 24 h was 86.01% and 80.18%, respectively. Additionally, chrysin at the concentration of 5, 10, 25, and 50 µM significantly inhibited NO production in THP-1 human monocytic cell line. Conclusions : Chrysin might have anti-oxidative activity related to its inhibition of hydrogen peroxide production in 'poly-IC and lipoteichoic acid'-stimulated RAW 264.7.

• Journal of Radiation Protection and Research
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• 제22권3호
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• pp.153-160
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• 1997

사람의 말초혈액림프구와 C57BL/6마우스의 비장림프구를 사용하여 시험관내에서 감마선을 조사하고 배양하여 세포질분열 차단 림프구내에 형성되는 미소핵의 빈도를 측정하였다. 미소핵 발생빈도는 방사선조사 선량에 비례하여 증가하였으며 linen-quadratic 곡선식에 적용하여, 세포 당 0.2개의 미소핵이 유도되는 방사선량을 산출하면 사람의 말초혈액 림프구에 비하여 마우스 비장림프구에서 1.67배 민감하였다. 미소핵시험방법을 이용하여, 사람의 말초혈액 림프구에 대한 인삼의 방사선 방호효과를 시험관내 시험으로, 마우스의 미장림프구에 대한 효과를 생체내 시험으로 검정하였다. 사람림프구에 있어서 방사선(3Gy)에 의해 유도되는 미소핵의 수는 방사선조사 전 및 후 투여군에서 공히 감소하였으며(p<0.01), 마우스를 사용한 생체시험에서도 림프구의 미소핵 발생빈도는 낮게 관찰되었다(p<0.025). 이상의 결과에서 인삼은 인체에서도 방사선에 의한 세포장해를 감소시킬 가능성을 나타냈다.

• Archives of Pharmacal Research
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• 제19권4호
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• pp.251-257
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• 1996

The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is subject of great concern at present. In this respect, the genetic toxicity of fenpropathrin ((RS)-.alpha.-cyano-3-phenoxybenzyl-2,2,3,3-tetramethyl cyclopropane carboxylate, CAS No.:39514-41-8), a pyrethroid insecticide, was evaluated in bacterial gene mutation system, chromosome aberration in mammalian cell system and in vivo micronucleus assay with rodents. In bacterial gene mutation assay, no mutagenicity of fenpropathrin (62-$5000\mug/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activaton system. In mammalian cell system using chinese hamster lung fibroblast, no clastogenicity of fenpropathrin was also observed both in the absence and in the presence of metabolic activation system in the concentration range of $7-28\mug/ml$. And also, in vivo micronucleus assay using mouse bone marrow cells, fenpropathrin also revealed no mutagenic potential in the dose range of 27-105 mg/kg body weight of fenpropathrin (i.p.). Consequently, no mutagenic potential of fenpropathrin was observed in vitro bacterial, mammalian mutagenicity systems and in vivo micronucleus assay in the dose ranges used in this experiment.

• 생명과학회지
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• 제24권5호
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• pp.485-490
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• 2014

백복령(Poria cocas)은 전통적으로 민간에서 피부미백에 효과가 있다고 보고되어 있다. 지금까지 melanin 합성에 대한 백복령의 직접적인 효과는 과학적으로 잘 연구되어 있지 않다. 따라서 멜라닌 합성에 대한 백복령 주정 추출물(PCEE)의 직접적인 효과를 밝히기 위해, 쥐의 B16F1 세포를 이용하여 DOPA synthesis assay, tyrosinase activity assay, Western blotting for melanogenic proteins [tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2]를 수행하였다. 본 연구에서 PCEE가 3,4-dihydroxyindole-2-carboxylic acid (DOPA) 합성을 차단함으로써 농도 의존적으로 melanin형성을 억제한다는 것이 밝혀졌다. 비록 tyrosinase의 활성은 영향을 받지 않았지만 TRP-1 과 TRP-2의 단백질 발현 수준은 PCEE에 의해 조절되었다. 따라서 이러한 결과는 PCEE가 미백 효능을 가지고 있어 미백 화장품 개발을 위해 이용될 수 있다는 것을 시사하고 있다.

• 한국수산과학회지
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• 제50권5호
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• pp.547-552
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• 2017

The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.