• Environmental Analysis Health and Toxicology
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• 제20권1호
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• pp.87-95
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• 2005

Cadmium is well known as a toxic metal and has insulin mimicking effects in rat adipose tissue. This study was undertaken to investigate the effect of CdCl₂ on glucose transport and its mechanism in 3T3 - L1 adipocytes. CdCl₂ exhibits respectively 2.2 and 2.8 fold increases in the 2-deoxyglucose uptake when exposed to 10 and 25 μM of CdCl₂ for 12 hr. To investigate the stimulating mechanism of glucose transport induced by CdCl₂. Wortmannin and PD98059 were used respectively as PI3K inhibitor and MAPK inhibitor, which did not affect 2-DOG uptake. This results suggest that induced 2-deoxy-(l-3H)-D-glucose (2-DOG) uptake by CdCl₂ may not be concerned with the insulin signalling pathway. Whereas nifedipine, a calcium channel blocker inhibited the 2- DOG uptake stimulated by CdCl₂. In addition, we also measured the increased production of Reactive oxygen substances (ROS) and glutathione (GSH) level in 3T3-L1 adipocytes to investigate correlation between the glucose uptake and increased production of ROS with H2DCFDA. CdCl₂ increased production of ROS. Induced 2-DOG uptake and increased production of ROS by CdCl₂ were decreased by N-acetylcystein (NAC). And L-buthionine sulfoximine (BSO) a potent inhibitor of γ-GCS, decreased of 2-DOG uptake. Also NAC and BSO changed the cellular GSH level, but GSH/GSSG ratio remained unchanged at 10, 25 μM of CdCl₂.

• ALGAE
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• 제29권4호
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• pp.311-319
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• 2014

The biocide sodium hypochlorite (NaOCl) is widely used for controlling algal growth, and this application can be extended to marine environments as well. This study evaluates the biocidal efficiency and cellular toxicity of NaOCl on the harmful dinoflagellate Cochlodinium polykrikoides, with emphasis on pigment production and antioxidant enzyme activity. The test organism showed dose-dependent decrease in growth rate on exposure to NaOCl, and the 72 h $EC_{50}$ was measured to be $0.584mg\;L^{-1}$. NaOCl significantly decreased pigment levels and chlorophyll autofluorescence intensity, indicating possible detrimental effects on the photosystem of C. polykrikoides. Moreover, it significantly increased the activities of antioxidant enzymes, suggesting the production of reactive oxygen species in the cells. These data indicate that NaOCl exerted deleterious effects on the photosynthetic machinery and induced oxidative damage in the dinoflagellate and this biocide could be effectively used for the control of algal blooms.

• Animal Bioscience
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• 제36권5호
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• pp.710-719
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• 2023

Objective: The present study investigated whether protodioscin (PD), a steroidal saponin mainly found in rhizome of Dioscorea species, alleviates oxidative stress-induced damage of porcine oocytes during in vitro maturation. Methods: Oocytes were treated with different concentrations of PD (0, 1, 10, 100, and 200 µM) in the presence of 200 µM H₂O₂ during in vitro maturation. Following maturation, spindle morphology and mitogen-activated protein kinase activity was assessed along with reactive oxygen species level, GSH activity, and mRNA expression of endogenous antioxidant genes at the MII stage. On the day 7 after parthenogenetic activation, blastocyst formation rate was calculated and the quality of embryo and mRNA expression of development-related genes was evaluated. Results: Developmental competence was significantly poorer in the 0 µM PD-treated (control) group than in the non-treated (normal) and 10 µM PD-treated (10PD) groups. Although the reactive oxygen species level did not significantly differ between these three groups, the glutathione level and mRNA expression of antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, nuclear factor erythroid 2-related factor 2 [Nrf2], and hemo oxygenase-1 [HO-1]) were significantly higher in the normal and 10PD groups than in the control group. In addition, the percentage of oocytes with defective spindle and abnormal chromosomal alignment was significantly lower and the ratio of phosphorylated p44/42 to total p44/42 was significantly higher in the normal and 10PD groups than in the control group. The total cell number per blastocyst was significantly higher in the 10PD group than in the control group. The percentage of apoptotic cells in blastocysts was highest in the control group; however, the difference was not significant. mRNA expression of development-related genes (POU domain, class 5, transcription factor 1 [POU5F1], caudal type homeobox 2 [CDX2], Nanog homeobox [NANOG]) was consistently increased by addition of PD. Conclusion: The PD effectively improves the developmental competence and quality of blastocysts by protecting porcine oocytes against oxidative stress.

• Food Science and Biotechnology
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• 제14권4호
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• pp.479-486
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• 2005

Arabinoxylan, a complex polysaccharide in cereal cell walls, has recently received research attention as a biological response modifier. The immunomodulating effect of arabinoxylans from rice bran (AXrb) was studied using a combined process of extrusion and commercial hemicellulase treatment in order to elucidate the augmentation mechanism of cell-mediated immunity in vitro. The cytotoxicity of mouse spleen lymphocytes against YAC-1 tumor cells was significantly enhanced by treatment with AXrb at $10-100\;{\mu}g/mL$. In an attempt to investigate the mechanism by which AXrb enhance NK cytotoxicity, we examined the effect of AXrb on cytokine production by spleen lymphocytes. Culture supernatants of the cells incubated with AXrb were collected and analyzed for IL-2 and IFN-${\gamma}$ synthesis by ELISA. IL-2 and IFN-${\gamma}$ production were increased significantly. These results suggest that AXrb may induce Th1 immune responses. Macrophages play an important role in host defenses against tumors by killing them and producing secretory products, which protect against bacterial, viral infection and malignant cell growth. AXrb were examined for their ability to induce secretory and cellular responses in murine peritoneal macrophages. When macrophages were treated with various concentrations ($10-100\;{\mu}g/mL$) of AXrb, AXrb induced tumoricidal activity, as well as increasing phagocytosis and the production of NO, $H_2O_2$, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These results indicate that reactive oxygen species, reactive nitrogen species, and inflammatory cytokines are likely to be the major mediators of tumoricidal activity in AXrb-treated macrophages. Therefore, AXrb may be useful in cancer immunotherapy and it is anticipated that AXrb obtained using extrusion and subsequent enzyme treatment can be used as an ingredient in nutraceuticals and cereal-based functional food.

• 대한본초학회지
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• 제28권3호
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• pp.107-113
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• 2013

Objective : The purpose of this study was to investigate the effect of fermented Artemisiae Argyi Folium(AAF) on some activities of human hepatoma cell, HepG2. Method : To investigate the effect of fermented Artemisiae Argyi Folium(AAF) activity on the human hepatoma cells, AAF extracts was fermented by Lactobacillus pentosus K34(AFL) and Sacchromyces cerevisiae STV89(AFS). And the effects of AFL or AFS on the activities of HepG2 cell, such as cell viability, nitric oxide(NO) production and reactive oxygen species(ROS) production, were tested. Result : Human Hepatoma Cells were incubated each for 3 hours and 24 hours. Human Hepatoma Cells treated with the extract was measured with MTT assay. Then AFL was found to be non-toxic at concentrations of 10 ug/mL(3h), 100 ug/mL(24h) or more. AFS was the same result at concentrations of more than 10 ug/mL. The extract increased ROS generation in Human Hepatoma Cells. AFL increased at concentrations of 100 ug/mL more (3h, also 10 ug/mL more) and 50 ug/mL(24h) and AFS increased both 50 ug/mL. In point of NO generation, AFL inhibited at concentrations of 10 ug/mL(3h) and 100 ug/mL(24h) more (3h, also 10 ug/mL more) and AFS also inhibited 50 ug/mL or more. Conclusion : AFL and AFS, obtained from Artemisiae Argyi Folium extracts by fermentation, reduced the NO production and increased ROS production in HepG2 cell, without cytotoxicity on HepG2 cell. The results suggested that AFL and AFS increased the immunological effects of Artemisiae Argyi Folium extracts.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.23-28
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• 2023

코로나바이러스-19로 인한 세계적인 펜데믹 이후 면역력 강화소재에 대한 관심이 급격히 증가하고 있으므로 산업적, 건강적 측면에서 새로운 소재의 개발이 필요하다. 본 연구에서는 연자육 추출물을 소재로 선정하고 RAW 264.7 쥐 대식세포를 이용하여 면역증진 효과를 평가하였다. 연자육 추출물은 RAW 264.7 세포에서 세포 생존력에 독성을 나타내지 않으면서 nitric oxide 및 reactive oxygen species의 생산을 상향 조절하였다. 또한 연자육 추출물은 RAW 264.7 세포에서 inducible nitric oxide synthase 및 cyclooxygenase-2 발현을 크게 증가시켰다. Enzyme-linked immunosorbent assay 결과에서는 연자육 추출물의 처리가 RAW 264.7 세포에서 interleukin 6 및 tumor necrosis factor-α의 생성을 유의미하게 향상시키는 것으로 나타났다. 또한 연자육 추출물이 p65, I kappa B kinase α/β, 및 I kappa B (IκB) α의 인산화를 크게 상향 조절하고, RAW 264.7 세포에서 IκB α의 발현을 하향 조절하였다. 우리의 연구 결과는 연자육 추출물이 Nuclear factor-kappa B 신호전달 경로를 통해 면역력을 향상시킬 수 있는 잠재적인 건강기능식품 소재가 될 수 있음을 나타낸다.

• Restorative Dentistry and Endodontics
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• 제48권1호
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• pp.6.1-6.14
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• 2023

Objectives: This study evaluated the effects of high-plasticity mineral trioxide aggregate (MTA-HP) on the activity of M1 and M2 macrophages, compared to white MTA (Angelus). Materials and Methods: Peritoneal inflammatory M1 (from C57BL/6 mice) and M2 (from BALB/c mice) macrophages were cultured in the presence of the tested materials. Cell viability (MTT and trypan blue assays), adhesion, phagocytosis, reactive oxygen species (ROS) production, and tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β production were evaluated. Parametric analysis of variance and the non-parametric Kruskal-Wallis test were used. Results were considered significant when p < 0.05. Results: The MTT assay revealed a significant decrease in M1 metabolism with MTA-HP at 24 hours, and with MTA and MTA-HP later. The trypan blue assay showed significantly fewer live M1 at 48 hours and live M2 at 48 and 72 hours with MTA-HP, compared to MTA. M1 and M2 adherence and phagocytosis showed no significant differences compared to control for both materials. Zymosan A stimulated ROS production by macrophages. In the absence of interferon-γ, TNF-α production by M1 did not significantly differ between groups. For M2, both materials showed higher TNF-α production in the presence of the stimulus, but without significant between-group differences. Likewise, TGF-β production by M1 and M2 macrophages was not significantly different between the groups. Conclusions: M1 and M2 macrophages presented different viability in response to MTA and MTA-HP at different time points. Introducing a plasticizer into the MTA vehicle did not interfere with the activity of M1 and M2 macrophages.

• Restorative Dentistry and Endodontics
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• 제45권4호
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• pp.54.1-54.16
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• 2020

Objectives: This study aimed to synthesize nanocrystals (NCs) of zinc oxide (ZnO) and calcium ion (Ca²⁺)-doped ZnO with different percentages of calcium oxide (CaO), to evaluate cytotoxicity and to assess the effects of the most promising NCs on cytotoxicity depending on lipopolysaccharide (LPS) stimulation. Materials and Methods: Nanomaterials were synthesized (ZnO and ZnO:xCa, x = 0.7; 1.0; 5.0; 9.0) and characterized using X-ray diffractometry, scanning electron microscopy, and methylene blue degradation. SAOS-2 and RAW 264.7 were treated with NCs, and evaluated for viability using the MTT assay. NCs with lower cytotoxicity were maintained in contact with LPS-stimulated (+LPS) and nonstimulated (-LPS) human dental pulp cells (hDPCs). Cell viability, nitric oxide (NO), and reactive oxygen species (ROS) production were evaluated. Cells kept in culture medium or LPS served as negative and positive controls, respectively. One-way analysis of variance and the Dunnett test (α = 0.05) were used for statistical testing. Results: ZnO:0.7Ca and ZnO:1.0Ca at 10 ㎍/mL were not cytotoxic to SAOS-2 and RAW 264.7. +LPS and -LPS hDPCs treated with ZnO, ZnO:0.7Ca, and ZnO:1.0Ca presented similar NO production to negative control (p > 0.05) and lower production compared to positive control (p < 0.05). All NCs showed reduced ROS production compared with the positive control group both in +LPS and -LPS cells (p < 0.05). Conclusions: NCs were successfully synthesized. ZnO, ZnO:0.7Ca and ZnO:1.0Ca presented the highest percentages of cell viability, decreased ROS and NO production in +LPS cells, and maintenance of NO production at basal levels.

• Toxicological Research
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• 제18권2호
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• pp.117-127
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• 2002

Saururus chinensis Baill has been used in Korean folk medicine for the treatment of various diseases such as edema, Jaundice, and furuncle. The components of this plant were extracted into four fraction. Among the four fraction, hexane and ethyl acetate fraction were highly toxic to 3T3 mouse embryo fibroblast and Raw 264.7 mouse macrophage, but n-butanol and residue fraction did not show any toxic effect to those cell lines. n-Butanol and residue fraction exhibited antioxidant effects on hydro-gen peroxide, hydroxyl radical, and superoxide anion directly in vitro and in the 3T3 fibroblasts. All the four fractions inhibited lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS) formation. In addition, n-butanol and residue fraction showed inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide production, and also down-regulated inducible nitric oxide synthase (iNOS) mRNA transcription 6 h after LPS stimulation in Raw 204.7 cells. Only n-butanol fraction, which mainly consists of flavonoids, inhibited NF-kB activation by decreasing IkBa degradation 90 min after LPS stimulation. horn the results, it is suggested that this plant could be a good candidate material for drug development based on its antioxidant and/or anti-inflammatory constituents.

• 한국식품저장유통학회지
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• 제18권1호
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• pp.65-71
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• 2011

본 연구에서는 도라지 분말을 유기용매로 추출한 후 도라지의 추출물과 분획물들에 의한 세포 내 활성산소종 및 glutathione (GSH)를 측정하여 항산화효과를 검토하였고 NO 생성 저해 효과를 알아보았다. 세포 내 활성산소종 생성억제 실험에서 건조 도라지의 A+M 및 MeOH 추출물과 추출물을 n-hexane, 85% aq. MeOH, n-BuOH, water로 다시 추출하여 얻어진 각각의 분획물들을 농도별로 HT1080 세포에 처리하였을 때 A+M과 MeOH 추출물 모두 측정시간 120분 동안 $500\;{\mu}M$ $H_2O_2$만을 처리한 control군에 비해 세포 내 활성산소종을 크게 억제시켰으며, MeOH 추출물에 의한 항산화 효과가 더 높게 나타났다. 또한, 각 분획물들 중 n-BuOH 분획물이 다른 분획물들에 비해 우수한 항산화 활성을 보였다. GSH 농도 측정 실험에서 A+M 및 85% aq. MeOH 분획물를 처리했을 때 GSH 함량이 증가하였다. NO 생성 저해 실험에서는 A+M 및 MeOH 추출물이 0.01 및 0.05 mg/mL의 농도에서 NO 생성을 억제하는 것으로 나타났으며, A+M 추출물에 의한 저해 효과가 높았다. 각 분획물들은 모두 control보다 낮은 NO 생성량을 나타내었으며, 특히 85% aq. MeOH 및 n-BuOH 분획물은 0.05 mg/mL 농도에서 blank에 가까운 NO 생성 억제율을 나타냈다. 이상의 연구결과로부터 n-BuOH 분획물에 의한 세포 내 활성산소종 생성 억제 효과 및 NO 생성 저해 효과가 우수함을 알 수 있었으며, 향후 분획물의 분리 정제를 통한 새로운 기능성 물질의 개발이 필요할 것으로 사료된다.