• Reproductive and Developmental Biology
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• 제39권1호
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• pp.1-6
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• 2015

Luteolysis is a cyclical regression of the corpus luteum in many non-primate mammalian species. Prostaglandin $F_2{\alpha}$($PGF_2{\alpha}$) from the uterus and ovary induces functional and structural luteolysis in bovine. The action of $PGF_2{\alpha}$ is mediated by $PGF_2{\alpha}$ receptor located on the luteal steroidogenic and endothelial cell membranes. $PGF_2{\alpha}$ plays an important role in regulating nitric oxide production in endothelial cells of the bovine corpus luteum. Nitric oxide production and nitric oxide synthase activity are stimulated and induced by $PGF_2{\alpha}$ in luteal endothelial cells. Moreover, the reactive oxygen species inhibits progesterone secretion in bovine luteal cells and induces apoptosis. Thus, the interaction between $PGF_2{\alpha}$ and reactive oxygen species provides important aspects in physiology of the corpus luteum forfunctional and structural luteolysis.

• Environmental Analysis Health and Toxicology
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• 제11권1_2호
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• pp.49-57
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• 1996

The production of reactive oxygen species on addition of hexavalent chromium (potassium dichromate, $K_2Cr_2O_7$ ) to lung cells in culture was studied using flow cytometer analysis. A Coulter Epics Profile flow cytometer was used to detect the formation of reactive oxygen species after $K_2Cr_2O_7$ was added to A549 cells grown to confluence. The cells were loaded with the dye, 2',7'-dichlorofluorescein diacetate, after which cellular esterases removed the acetate groups and the dye was trapped intracellularly. Reactive oxygen species oxidized the dye, with resultant fluorescence. Increased doses of Cr(VI) caused increasing fluorescence (10-fold higher than background at 200 gM). Addition of Cr(III) compounds, as the picolinate or chloride, caused no increased fluorescence. Electron paramagnetic resonance (EPR) spectroscopic studies indicated that three (as yet unidentified) spectral "signals" of the free radical type were formed on addition of 20, 50, 100 and 200 gM Cr(VI) to the A549 cells in suspension. Two other EPR 'signals" with the characteristics of Cr(V) entities were seen at field values lower than the standard free radical value. radical value.

• 약학회지
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• 제52권6호
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• pp.441-445
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• 2008

To investigate effect of caffeic acid on the intracellular reactive oxygen species production, we used DHE for intracellular superoxide anion production, DCF for intracellular ${H_2}{O_2}$ production and DHR for intracellular hydroperoxide production in Raw 264.7 cells. DPPH assay showed that antioxidant activity of caffeic acid with 39.5 ${\mu}M$ of ${IC}_{50}$ values was similar to that of ascorbic acid with 41.3 ${\mu}M$ of ${IC}_{50}$ values. Caffeic acid dose-dependently inhibited silica-induced ${H_2}{O_2}$ and hydroperoxide production but did not affect superoxide anion production in Raw 264.7 cells, which suggest that antioxidant effect of caffeic acid acts on the post-step of superoxide anion. On the other hand, caffeic acid showed a potent antioxidant effect in $lCuSO_4$-induced lipid peroxidation. Furthermore, plasma superoxide dismutase activity (3.43${\pm}$0.23 U/ml) in 10 mg/kg caffeic acid-fed mice was significantly higher than that (2.32${\pm}$0.24 U/ml) of control. From the above results, it is referred that caffeic acid appears to have potent anti-oxidant activity in both cell system and in vivo system.

• Journal of Periodontal and Implant Science
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• 제39권3호
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• pp.331-337
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• 2009

Purpose: Interleukin (IL)-8 is one of pro-inflammatory cytokines. Reactive oxygen species (ROS) are reduced metabolites of $O_2$. Aggregatibacter actinomycetemcomitans is one of representative periodontopathogens. To investigate the role of A. actinomycetemcomitans in IL-8 expression of periodontal ligament (PDL) cells, we estimated the production of IL-8 and ROS in A. actinomycetemcomitans treated PDL cells. Methods: The IL-8 production was determined by enzyme-linked immunosorbent assay. The ROS production was estimated using H2DCFDA and FACS. Results: A. actinomycetemcomitans increased the production of IL-8 and ROS at 10, 100, and 500 multiplicity of infection. N-acetylcysteine, an antioxidant of ROS, down-regulated the production of IL-8 induced by A. actinomycetemcomitans. Conclusions: These results suggest that A. actinomycetemcomitans induces IL-8 production and ROS may act as a mediator in this process.

• Biomolecules & Therapeutics
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• 제20권2호
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• pp.165-170
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• 2012

Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

• 생약학회지
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• 제48권4호
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• pp.273-279
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• 2017

The aim of this study was to investigate the mechanisms underlying apoptosis induced by a broussochalcone B (BCB) from Broussonetia papyrifera in HepG2 cells. The results showed that BCB treatment for 24 hr significantly inhibited cell viability in a dose-dependent manner, and induced apoptosis in HepG2 cells. More so, BCB treatment triggered the cleavage of caspase-8, -9, -3, poly (ADP-ribose) polymerase (PARP), increase of Bax level, and decrease of Bcl-2 expression. A general caspase inhibitor (z-VAD-fmk) blocked BCB-induced cell death. Furthermore, BCB treatment caused reactive oxygen species (ROS) production in a dose-dependent manner. In addition, an antioxidant N-acetylcysteine (NAC) blocked BCB-induced ROS production and cell death. Therefore, these results indicate that BCB-induced apoptosis is mediated by a caspase dependent pathway and ROS production in HepG2 cells.

• 대한의생명과학회지
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• 제23권2호
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• pp.128-132
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• 2017

Streptozotocin (STZ), bisphenol A (BPA), and diethylstilbestrol (DES) are known as endocrine disruptors, occurs oxidative stress in animal cells. Generally, oxidative stress induces reactive oxygen species (ROS) and lipid peroxidation of sperm and lead to decreased viability and fertility in pigs. Therefore, we investigated the influence of STZ, BPA and DES on ROS production and lipid peroxidation on boar sperm. Collected sperm were incubated with semen extender containing $10{\mu}M\;STZ$, $10{\mu}M\;BPA$ and $20{\mu}M\;DES$ for 3, 6 and 9 hours. Intracellular ROS and lipid peroxidation of sperm were analyzed by 2', 7'-dichlorofluorescein diacetate and malondialdehyde methods. The results show that, intracellular ROS was not significantly different among the all treatments, but lipid peroxidation was significantly increased in STZ group at 3 hour after incubation with boar sperm (P<0.05). These results suggest that STZ stimulates lipid peroxidation more than ROS production and may exert a negative effect on sperm fertility.

• 대한수의학회지
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• 제60권2호
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• pp.79-83
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• 2020

Fenbendazole (FBZ) is a benzimidazole anthelmintic that has been widely used in treatments for gastrointestinal parasites including pinworms and roundworms in animals. Recently, some studies demonstrated that FBZ has anti-cancer effects related to disruption of microtubule polymerization. In this study, we investigated whether FBZ has anti-cancer activity in HL-60 cells, a human leukemia cell line, and assessed its relationship with the production of reactive oxygen species (ROS). FBZ treatment at 0.25-1 μM significantly decreased the metabolic activity of HL-60 cells. The mitochondrial membrane potential of FBZ-treated HL-60 cells decreased in a concentration-dependent manner. Apoptosis analysis using annexin V-FITC/propidium iodide staining demonstrated that 1 μM FBZ increased the percentages of cells in apoptosis and necrosis. In addition, Hoechst 33342 staining showed the presence of broken nuclei in HL-60 cells treated with 0.5 and 1 μM FBZ. To investigate the anti-cancer mechanism of FBZ, HL-60 cells were treated with FBZ in the absence or presence of N-acetyl cysteine (NAC), an inhibitor of ROS production. NAC significantly recovered the decreased metabolic activity of HL-60 induced by 0.5 and 1 μM FBZ treatments. This study provides evidence that FBZ has anti-cancer activity in HL-60 cells provided, in part, via ROS production.

• 한국임상수의학회지
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• 제26권1호
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• pp.8-16
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• 2009

항암제에 의해 흰쥐에서 다형핵 백혈구(polymorphonuclear leukocytes, PMN) 의 활성산소종(reactive oxygen species, ROS)과 산화질소(nitric oxide, NO)의 생성변화에 대해 연구하였다. 최근 자극유도인자에 의한 PMN의 호흡방출은 protein kinase C (PKC)의 활성, inositol phosphate transduction pathway의 활성, 그리고 intracellular [$Ca^{2+}$]와 관계가 있다고 밝혀졌으며, 본 연구에서 사용된 항암제(cyclophosphamide, cisplatin, tamoxifen, doxifluridine)중 일부는 화학치료제로써 비특이적으로 면역을 억제하는데 사용되고 있다. 암 치료 시 백혈구의 방어기능에 미치는 영향을 연구하기 위한 목적으로 in vitro에서 각 항암제를 처리한 PMN을 배양하여 ROS와 NO의 생성변화와 이차적 신호전달계인 phospholipase C(PLC), D(PLD), PKC, tyrosine phosphorylation kinase (TPK)와 phosphatidylinositol-3 kinase의 억제율을 측정하였다. PMN에 각각cyclophosphamide, cisplatin, tamoxifen, doxifluridine을 short term(${\leq}4hrs$) 처리시, formylmethionyl-leucy1-phenylalanine (FMLP) 자극에 의해 호흡방출의 증가가 나타났다. 반면, long term (8hrs) 처리 시, ROS의 생성은concentration-dependent 방법으로 감소되었다.

• Reproductive and Developmental Biology
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• 제38권1호
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• pp.1-8
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• 2014

Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxygen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.