• 제목/요약/키워드: reactive oxygen species production

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Analysis of Nutritional Components and Cell-based Antioxidant Activity on Brassica juncea Cultivated in Jeongseon, South Korea (정선 갓의 영양성분 및 세포기반 항산화 활성 분석)

  • Kwon, Hee-Yeon;Choi, Sun-Il;Cho, Bong-Yeon;Choi, Seung-Hyun;Sim, Wan-Sup;Han, Xionggao;Jang, Gill-Woong;Choi, Ye-Eun;Yeo, Jin-Hui;Cho, Ju-Hyun;Lee, Ok-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.32 no.5
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    • pp.462-472
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    • 2019
  • The purpose of this study was to demonstrate the quality characteristics of Brassica juncea cultivated in Jeongseon (BJJ), South Korea. We analyzed the nutritional components and antioxidant activity of BJJ. As a result of the free sugar analysis, the contents of glucose and fructose in BJJ were $0.29{\pm}0.02g/100g$ and $0.10{\pm}0.00g/100g$, respectively. The major fatty acids were palmitic acid, octadecenoic acid and stearic acid. The palmitic acid was the highest at 31.22% of all fatty acids. The major minerals were identified as Ca, P, K, Mg and Na. The contents of vitamin $B_1$, vitamin $B_2$, vitamin $B_6$, vitamin C and vitamin E in BJJ were $0.02{\pm}0.00mg/100g$, $0.087{\pm}0.01mg/100g$, $0.02{\pm}0.00mg/100g$, $0.56{\pm}0.06mg/100g$ and $0.20{\pm}0.03mg\;{\alpha}-TE/100g$, respectively. As a result of the free amino acid analysis, total amino acid contents in BJJ were $2,801.21{\pm}115.38mg/100g$. L-proline content was the highest ($744.30{\pm}119.06mg/100g$) in BJJ. BJJ extract inhibits reactive oxygen species production in 3T3-L1 adipocytes. Also, BJJ extract exhibits a protective effect on oxidative stress in $H_2O_2$-induced human dermal fibroblast. These results indicate that BJJ comprises various valuable nutrients which can be used as functional food ingredients.

The enhancing effect of Acanthopanax sessiliflorus fruit extract on the antibacterial activity of porcine alveolar 3D4/31 macrophages via nuclear factor kappa B1 and lipid metabolism regulation

  • Hwang, Eunmi;Kim, Gye Won;Song, Ki Duk;Lee, Hak-Kyo;Kim, Sung-Jo
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1776-1788
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    • 2019
  • Objective: The demands for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). Methods: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells ($M{\Phi}$) were activated by phorbol 12-myristate 13-acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B ($NF-{\kappa}B$) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative polymerase chain reaction was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 $M{\Phi}$ cells was evaluated by the colony forming unit assay. Results: ASF upregulated the cell viability and growth rate of 3D4/31 $M{\Phi}$, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of $NF-{\kappa}B$ protein, tumor necrosis factor $(TNF){\alpha}$ mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of $NF-{\kappa}B$, $TNF{\alpha}$, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 $M{\Phi}$ against Escherichia coli. Conclusion: This study provides a novel insight into the regulation of $NF-{\kappa}B$ activity and lipid metabolism in $M{\Phi}$, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

Cellular protective effect of Ecklonia cava extract on ultra-fine dust (PM2.5)-induced cytoxicity (초미세먼지(PM2.5)로 유도된 in vitro 세포 독성에 대한 감태(Ecklonia cava) 추출물의 보호 효과)

  • Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Yoo, Seul Ki;Han, Hye Ju;Shin, Eun Jin;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.503-508
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    • 2019
  • To evaluate the protective effect of Ecklonia cava on ultra-fine dust ($PM_{2.5}$)-induced cytotoxicity, we investigated the in vitro antioxidant activity and cell viability after exposure to $PM_{2.5}$. E. cava was extracted using water and 80% ethanol, and antioxidant activity was determined using the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS)/2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and lipid peroxidation inhibition assays. The 80% ethanol extract showed relatively higher antioxidant activity than the water extract. The cell protective effects were determined by measuring the intracellular reactive oxygen species (ROS) content and viability of nasal epithelial (RPMI-2650), lung epithelial (A549), and brain neuroblastoma (MC-IXC) cells. Results showed that the 80% ethanol extract inhibited ROS production more than the water extract. In contrast, both extracts showed similar effects on cell viability in the $PM_{2.5}$-induced cell death assay. Thus, Ecklonia cava may act as an effective resource for preventing $PM_{2.5}$-induced cytotoxicity in nasal, lung, and brain cells.

The Inhibitiory Effects of the Scirpi rhizoma on Melanogenesis (삼릉추출물의 멜라닌 생성 저해 효과)

  • Lee, Bum-Chun;Kim, Jin-Hwa;Sim, Gwan-Sub;Zhang, Yong-He;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.305-310
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    • 2005
  • To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

Anti-Oxidative and Inhibitory Effect of Saussurea involucrata on MMP-1 in UVA-irradiated Human Dermal Fibroblast (설련의 항산화능와 사람 섬유아세포에서 UVA에 의한 MMP-1발현 저해효과)

  • Sim, Gwan-Sub;Kim, Jin-Hwa;Na, Young;Lee, Dong-Hwan;Lee, Bum-Chun;Zhang, Yong-He;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.329-335
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    • 2005
  • In order to investigate the effects of Saussurea involucrata on (relationship between) aging (and Saussurea involucrata), we examined the activities of antioxidation, in vitro MMP inhibition and UVA-induced MMP-1 expression in human dermal fibroblasts. S. involucrata showed scavenging activities radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $3.89{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $67.29{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. At the concentration of $1000{\mu}g/mL$, S. involucrata showed 93.27% inhibition on lipid peroxidation of linoleic acid. S. involucrata inhibited the activities of MMP-1 in a does-dependent manner and the $IC_{50}$ value calculated from semi-log plots was $97.18{\mu}g/mL$. Also, UVA induced MMP expression in human dermal fibroblasts was reduced 42.86% by treatment with S. involucrata, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore S. involucrata was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that S. involucrata might act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

Regulation of toll-like receptors expression in muscle cells by exercise-induced stress

  • Park, Jeong-Woong;Kim, Kyung-Hwan;Choi, Joong-Kook;Park, Tae Sub;Song, Ki-Duk;Cho, Byung-Wook
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1590-1599
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    • 2021
  • Objective: This study investigates the expression patterns of toll-like receptors (TLRs) and intracellular mediators in horse muscle cells after exercise, and the relationship between TLRS expression in stressed horse muscle cells and immune cell migration toward them. Methods: The expression patterns of the TLRs (TLR2, TLR4, and TLR8) and downstream signaling pathway-related genes (myeloid differentiation primary response 88 [MYD88]; activating transcription factor 3 [ATF3]) are examined in horse tissues, and horse peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNs) and muscles in response to exercise, using the quantitative reverse transcription-polymerase chain reaction (qPCR). Expressions of chemokine receptor genes, i.e., C-X-C motif chemokine receptor 2 (CXCR2) and C-C motif chemokine receptor 5 (CCR5), are studied in PBMCs and PMNs. A horse muscle cell line is developed by transfecting SV-T antigen into fetal muscle cells, followed by examination of muscle-specific genes. Horse muscle cells are treated with stressors, i.e., cortisol, hydrogen peroxide (H2O2), and heat, to mimic stress conditions in vitro, and the expression of TLR4 and TLR8 are examined in stressed muscle cells, in addition to migration activity of PBMCs toward stressed muscle cells. Results: The qPCR revealed that TLR4 message was expressed in cerebrum, cerebellum, thymus, lung, liver, kidney, and muscle, whereas TLR8 expressed in thymus, lung, and kidney, while TLR2 expressed in thymus, lung, and kidney. Expressions of TLRs, i.e., TLR4 and TLR8, and mediators, i.e., MYD88 and ATF3, were upregulated in muscle, PBMCs and PMNs in response to exercise. Expressions of CXCR2 and CCR5 were also upregulated in PBMCs and PMNs after exercise. In the muscle cell line, TLR4 and TLR8 expressions were upregulated when cells were treated with stressors such as cortisol, H2O2, and heat. Migration of PBMCs toward stressed muscle cells was increased by exercise and oxidative stresses, and combinations of these. Treatment with methylsulfonylmethane (MSM), an antioxidant on stressed muscle cells, reduced migration of PBMCs toward stressed muscle cells. Conclusion: In this study, we have successfully cultured horse skeletal muscle cells, isolated horse PBMCs, and established an in vitro system for studying stress-related gene expressions and function. Expression of TLR4, TLR8, CXCR2, and CCR5 in horse muscle cells was higher in response to stressors such as cortisol, H2O2, and heat, or combinations of these. In addition, migration of PBMCs toward muscle cells was increased when muscle cells were under stress, but inhibition of reactive oxygen species by MSM modulated migratory activity of PBMCs to stressed muscle cells. Further study is necessary to investigate the biological function(s) of the TLR gene family in horse muscle cells.

Mitochondrial Dysfunction and Cancer (미토콘드리아 기능 이상과 암)

  • Han, Yu-Seon;Jegal, Myeong-Eun;Kim, Yung-Jin
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1034-1046
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    • 2019
  • The mitochondria is the major cellular organelle of energy metabolism for the supply of cellular energy; it also plays an important role in controlling calcium regulation, reactive oxygen species (ROS) production, and apoptosis. Mitochondrial dysfunction causes various diseases, such as neurodegenerative diseases, Lou Gehrig's disease, cardiovascular disease, mental disorders, diabetes, and cancer. Most of the diseases are age-related diseases. In this review, we focus on the roles of mitochondrial dysfunction in cancer. Mitochondrial dysfunction induces carcinogenesis and is found in many cancers. The factors that cause mitochondrial dysfunction differ depending on the types of carcinoma, and those factors could cause cancer malignancy, such as resistance to therapy and metastasis. Mitochondrial dysfunction is caused by a lack of mitochondria, an inability to provide key substances, or a dysfunction in the ATP synthesis machinery. The main factor associated with cancer malignancy is mtDNA depletion. Mitochondrial dysfunction would leads to malignancy through changes in molecular activity or expression, but it is not known in detail which changes lead to cancer malignancy. In order to explore the relationship between mitochondrial dysfunction and cancer malignancy in detail, mitochondria dysfunctional cell lines are constructed using chemical methods such as EtBr treatment or gene editing methods, including shRNA and CRISPR/Cas9. Those mitochondria dysfunctional cell lines are used in the study of various diseases caused by mitochondrial dysfunction, including cancer.

Anti-Inflammatory Effect of Ethanol Extract from the Seeds of Arctium Lappa L. in Vascular Endothelial Cells (혈관내피세포에서 우방자(牛蒡子) 에탄올 추출물의 항염증 효과)

  • Lee, Yun-Jung;Yoon, Jung-Joo;Kim, Hye-Yoom;Ahn, You-Mee;Hong, Mi-Hyeon;Son, Chan-Ok;Na, Se-Won;Lee, Ho-Sub;Kang, Dae-Gill
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.3
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    • pp.20-31
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    • 2019
  • Objectives: The seeds from Arctium lappa have been considered for its various pharmacological properties, which include anti-carcinogenic, anti-inflammatory, anti-diabetic, and anti-viral activities. Methods: In the present study, we investigated the anti-inflammatory effect of the ethanol extract from the seeds of Arctium lappa L (EAL) on cytokine-induced vascular inflammation in human umbilical vein endothelial cells (HUVEC). Results: Pretreatment with EAL significantly decreased tumor necrosis factor alpha ($TNF-{\alpha}$)-induced cell adhesion molecules expression such as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial-selectin (E-selectin) in a dose-dependent manner. Cell adhesion assay showed that pretreatment with EAL suppressed HUVEC-monocyte adhesion by $TNF-{\alpha}$ over $1{\mu}g/ml$ concentration. We investigated the involvement of nuclear transcription factor kappa-B ($NF-{\kappa}B$) in $TNF-{\alpha}$-induced vascular inflammation. $NF-{\kappa}B$ p65 nuclear expression was induced by $TNF-{\alpha}$, however, pretreatment with EAL was attenuated that nuclear translocation. In cytoplasm, EAL was also attenuated $TNF-{\alpha}$-induced decrease of inhibitor of ${\kappa}B-{\alpha}$ ($I{\kappa}B-{\alpha}$) expression. Moreover, EAL significantly decreased $TNF-{\alpha}$-induced production of intracellular reactive oxygen species (ROS). Conclusions: Taken together, our findings suggest that seeds of Arctium lappa L could be a therapeutic herb for prevention of cardiovascular diseases throughout the inhibition of vascular endothelial inflammation.

Detrimental effects of lipopolysaccharides on maturation of bovine oocytes

  • Zhao, Shanjiang;Pang, Yunwei;Zhao, Xueming;Du, Weihua;Hao, Haisheng;Zhu, Huabin
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.8
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    • pp.1112-1121
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    • 2019
  • Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

Dronedarone hydrochloride enhances the bioactivity of endothelial progenitor cells via regulation of the AKT signaling pathway

  • Zhang, Jian;Le, Thi Hong Van;Rethineswaran, Vinoth Kumar;Kim, Yeon-Ju;Jang, Woong Bi;Ji, Seung Taek;Ly, Thanh Truong Giang;Ha, Jong Seong;Yun, Jisoo;Cheong, Jae Hun;Jung, Jinsup;Kwon, Sang-Mo
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.5
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    • pp.459-466
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    • 2021
  • Cardiovascular disease (CVD) and its complications are the leading cause of morbidity and mortality in the world. Because of the side effects and incomplete recovery from current therapy, stem cell therapy emerges as a potential therapy for CVD treatment, and endothelial progenitor cell (EPC) is one of the key stem cells used for therapeutic applications. The effect of this therapy required the expansion of EPC function. To enhance the EPC activation, proliferation, and angiogenesis using dronedarone hydrochloride (DH) is the purpose of this study. DH received approval for atrial fibrillation treatment and its cardiovascular protective effects were already reported. In this study, DH significantly increased EPC proliferation, tube formation, migration, and maintained EPCs surface marker expression. In addition, DH treatment up-regulated the phosphorylation of AKT and reduced the reactive oxygen species production. In summary, the cell priming by DH considerably improved the functional activity of EPCs, and the use of which might be a novel strategy for CVD treatment.