• Journal of fish pathology
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• v.7 no.1
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• pp.63-70
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• 1994

Nitric oxide(NO) has recently been shown to play an important role on central nervous system(CNS) function in mammals. It is synthesized from L-arginine by the enzyme NO synthase. In this study, we examined this enzyme's existence in CNS of rainbow trout(Oncorhynchus mykiss) and described the forms of microglia. astrocytes. and oligodendrocytes. Two forms of microglia are distributed in CNS. one resembling their mammalian counterpart(large microglia : LM). and the other comprising small microglia(SM) with very little cytoplasm. CNS contained astrocytes of a distinct type which form reticular network, but lack connections to capillaries. The oligodendrocyte was generally a much denser cell than the astrocyte. We have detected NOS($1.04{\pm}0.12\;pg/min/mg$) from rainbow trout CNS. It could be inhibited reversibly or irreversibly by $N^{G}MMA$ and EGTA. These result suggest that the formation of NO from L-arginine in CNS is calcium-dependent and a pathway of early evolutionary orgin.

• Korean Journal of Plant Resources
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• v.32 no.5
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• pp.433-441
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• 2019

This study describes a preliminary evaluation of the anti-inflammatory activity and anti-atopic activity of Phormium tenax leaf extracts. P. tenax leaf was extracted using 70% ethanol and then fractionated sequentially with n-hexane, methylene chloride, ethyl acetate, n-butanol. In order to effectively screen for anti-inflammatory agents, we first investigated the inhibitory effects of P. tenax leaf crude extracts and solvent fractions on production of pro-inflammatory factors[nitric oxide(NO), prostaglandin $E_2(PGE_2)$, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2)] and pro-inflammatory cytokines [tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, interleukin-6(IL-6) and $interleukin-1{\beta}(IL-1{\beta})$] in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells. In addition, we also evaluated of their inhibitory effect on the atopic dermatitis-like inflammatory markers such as macrophage-derived chemokine(MDC) and thymus and activation-regulated chemokine(TARC) in HaCaT cells. Among the five solvent fractions of P. tenax, methylene chloride and ethyl acetate fractions inhibited production of pro-inflammatory factors and pro-inflammatory cytokines in a dose dependent manner, respectively. These fractions were also showed inhibitory activity for MDC and TARC expression levels in $IFN-{\gamma}-stimulated$ HaCaT cells, respectively. These results suggest that P. tenax have significantly effects of anti-inflammatory activity and anti-atopic activity that might be beneficial for the topical treatment of inflammatory skin disorders.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.4
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• pp.49-65
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• 2011

Objectives: This study was performed to investigate the effects of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) on the monosodium iodoacetate(MIA) induced early state osteoarthritis in rats. Methods: Osteoarthritis was induced by injection of MIA(0.25 mg) into knee joints of rats. Osteoarthritis rats were divided into control(n=8) and treated=8 group respectively, Control group was taken distilled water and treated group was taken extracts of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) by orally for 20 days. Body weight was measured at 0, 5, 10, 15, 20 days after MIA injection. At the end of experiment, gross and histopathological examination on the articular cartilages of the knee joints were performed. Proteoglycan(PG) content of articular cartilages were analysed by safranine O staining method. The content of tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$ in synovial fluids were analysed by enzyme-inked inmunosorbent assay(ELISA) method. And also cycloxygenase-2(COX-2), matrix metalloproteinase 3(MMP-3), inducible nitric oxide synthase(iNOS), calpain immunochistochemical examination on the knee joints were performed. Results: PG content in articular cartilages of the treated group was significantly increased compared with control group. Histopathological osteoarthritic score of the treated group was significantly decreased compared with control group. $TNF-{\alpha}$ content in synovial fluids, expression of iNOS and calpain in synovial membrane of the treated group were significantly decreased compared with control group. But body weight, $1L-1{\beta}$ content in synovial fluids, expression of iNOS and MMP-3 of the treated group were not significantly changed compared with control group. Conclusions: On the basis of these results, we conclude that Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) has anti-arthritic effects on the MIA induced early stage osteoarthritis in rats.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.1
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• pp.7-15
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• 2004

Nitric oxide (NO) system has been implicated in a wide range of physiological functions in the nervous system. However, the role of NO in regulating the neural activity in the gustatory zone of nucleus tractus solitarius (NTS) has not been established. The present study was aimed to investigate the role of NO in the gustatory NTS neurons. Sprague-Dawley rats, weighing about 50 g, were used. Whole cell patch recording and immunohistochemistry were done to determine the electrophysiological characteristics of the rostral gustatory nucleus of the tractus solitaries and distribution of NO synthases (NOS). Neuronal NOS (nNOS) immunoreactivity was strongly detected along the solitary tract extending from rostral to caudal medulla. Resting membrane potentials of NTS neurons were $-49.2{\pm}2\;mV$ and action potential amplitudes were $68.5{\pm}2\;mV$ with a mean duration measured at half amplitude of $1.7{\pm}0.3\;ms$. Input resistance, determined from the response to a 150 ms, -100 pA hyperpolarizing current pulse, was $385{\pm}15\;M{\Omega}$, Superfusion of SNAP or SNP, NO donors, produced either hyperpolarization (68%), depolarization (5%), or no effect (27%). The hyperpolarization was mostly accompanied by a decrease in input resistance. The hyperpolarization caused by SNAP or SNP increased the time to initiate the first action potential, and decreased the number of action potentials elicited by current injection. SNP or SNAP also markedly decreased the number of firing neural discharges of the spontaneous NTS neural activity under zero current. Superfusion of L-NAME, a NOS inhibitor, slightly depolarized the membrane potential and increased the firing rate of NTS neurons induced by current injection. ODQ, a soluble guanylate cyclase inhibitor, ameliorated the SNAP-induced changes in membrane potential, input resistance and firing rates. 8-Br-cGMP, a non-degradable cell-permeable cGMP, hyperpolarized the membrane potential and decreased the number of action potentials. It is suggested that NO in the gustatory NTS has an inhibitory role on the neural activity of NTS through activating soluble guanylate cyclase.

• Food Science and Preservation
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• v.24 no.3
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• pp.413-420
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• 2017

This study was designed to examine the in vitro antioxidant and anti-inflammatory effects of red beet (Beta vulagaris) root. Red beet root was extracted using 70% ethanol and then fractionated sequentially with n-hexane, ethyl acetate and butanol. Antioxidative ability was evaluated by bioassays using total polyphenol contents and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid diammonium salt) radical scavenging activity. Ethyl acetate fraction of red beet root was best on total polyphenol contents ($37.02{\pm}0.37mg\;GAE/g$) and ABTS radical scavenging effects ($IC_{50}$ $42.9{\pm}9.5{\mu}g/mL$). For the anti-inflammatory activity in RAW264.7 cells, the hexane fraction showed the highest inflammatory effect. Dose response studies were performed to determine the inhibitory effect of hexane fraction of red beet root on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The hexane fraction of red beet root inhibited the NO and $PGE_2$ production and the protein level of iNOS and COX-2, and protein expression of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$), in a dose-dependent manner. These results suggest that red beet root has considerable potential as a functional food ingredient with antioxidative and anti-inflammatory effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.4
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• pp.427-436
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• 2018

In this study, we investigated anti-inflammatory and anti-bacterial constituents from Daucus carota var. sativa (carrot) areal parts. For the extract and solvent fractions, the anti-inflammatory activities were examined by measuring the nitric oxide (NO) production using LPS-stimulated RAW 264.7 cells. Among them, the ethyl acetate (EtOAc) fraction decreased the NO level in a dose-dependent manner. To elucidate further anti-inflammatory mechanisms, EtOAc fraction was evaluated by estimating their effects on the production of prostaglandin $E_2$ and pro-inflammatory cytokines as well as on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). As a result, the EtOAc fraction was determined to inhibit the production of $PGE_2$, IL-$1{\beta}$, IL-6 and reduce the iNOS, COX-2 protein expression. Upon the anti-bacterial tests using Staphylococcus epidermidis and Propionibacterium acnes, n-hexane (Hex) and EtOAc fractions showed the most potent activities. Three phytochemicals were isolated form the EtOAc fraction; diosmetin (1), diosmin (2), cynaroside (3). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including $^1H$ and $^{13}C$ NMR spectra, as well as comparison of the data to the literatures. Anti-inflammatory and anti-bacterial effects were studied for the isolates. All of the compounds (1 - 3) decreased the NO production, effectively. Also, compound 3 showed anti-bacterial activity on P. acnes. Based on these results, D. carota var. sativa extract could be potentially applicable as anti-inflammatory and anti-bacterial ingredients in cosmetic formulations.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.46-46
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• 2021

The first purpose of this study was to evaluate the scavenging capacity (SC) of DPPH and ABTS free radicals for ethanol extract (STR-E) and its active fractions from Sophora tonkinensis root (STR). Four different fractions from STR-E were prepared by using different types of solvents such as chloroform (STR-E-C), ethyl acetate (STR-E-EA), n-butanol (STR-E-B), and water (STR-E-W). STR-E-C showed the highest value of total phenolic content, while STR-E showed the highest value of total flavonoid and terpenoid content. In STR-E and its four fractions, STR-E-EA showed the strongest SC with the lowest SC50 values of the DPPH radicals and ABTS radicals. The second purpose of this study was to evaluate anti-inflammatory activity in the lipopolysaccharide (LPS)-induced RAW 264.7 macrophages treated with STR-E, STR-E-C, and STR-E-EA, respectively. No cytotoxic effect to RAW 264.7 cells was observed at 20 ~ 25 ㎍/ml of STR-E, 10 ㎍/ml of STR-E-C, and 5 ㎍/ml of the STR-E-EA, presenting cell viability values close to that of the untreated control (100%). STR-E, STR-E-C, and STR-E-EA significantly suppressed the LPS-induced nitric oxide (NO) in a dose-dependent manner. Results of reverse-transcription (RT)-qPCR analysis showed that the peak mRNA levels of IL-1β, TNF-α, iNOS, IL-6, and IL-10 were observed in the LPS-stimulated macrophages at 4 h, 2 h, 12 h, 12 h, and 12 h, respectively. The peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced in the LPS-stimulated macrophages co-treated with 20 ㎍/ml and 25 ㎍/ml of STR-E, respectively. In the case of IL-10, its peak mRNA level slightly increased without statistical significance. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 10 ㎍/ml and 20 ㎍/ml of STR-E-C, respectively. In contrast, the peak mRNA level of IL-10 significantly increased at 8 h. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 5 ㎍/ml and 10 ㎍/ml of STR-E-EA, respectively. In contrast, the peak mRNA level of IL-10 increased at 4 h. Taken together, our data indicated that STR-E, STR-E-C, and STR-E-EA activate macrophages to secrete both pro-inflammatory and anti-inflammatory cytokines.

• The Korea Journal of Herbology
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• v.35 no.1
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• pp.35-44
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• 2020

Objectives : The objective of this study was to investigate the therapeutic effect of Corni Fructus 30% ethanol extract (CFE) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods : The subjects were divided into 4 groups ; Normal group (N, n=10), MIA-induced osteoarthritis control group (Con, n=10), indomethacin 5 mg/kg treated group (INDO, n=10), CFE 200 mg/kg treated group (CFE, n=10). Blood and articulation tissues were collected after two weeks of drug administration. Oxidative stress was analyzed with reactive oxygen species (ROS), peroxynitrite (ONOO-). And the Nuclear factor erythroid-2 (Nrf2), heme oxygenase-1 (HO-1), superoxide dismutase (SOD), catalase, glutathione peroxidase-1/2 (GPx-1/2), Nuclear Factor Kappa B p65 (NF-κBp65), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNFα), interleukin-6 (IL-6), Interleukin 1β (IL-1β), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1) were investigated by western blot. Results : The administration of CFE showed a significant reduction of changes in relative hind paw weight distribution. Reactive oxygen species (ROS) and peroxy nitrite (ONOO-) levels of articulation tissues were significantly decreased in CFE compared to the control group. Western blot measurements of Nrf2, HO-1, SOD, catalase, GPx-1/2 showed that the CFE group was increased compared to the Con group. And western blot measurements of NF-κBp65, COX-2, iNOS, TNFα, IL-6, IL-1β showed that the CFE group was reduced compared to the Con group. Also CFE group decreased MMP-1 and increased TIMP-1. Conclusion : Based on the above results, it can be seen that osteoarthritis is improved when Corni Fructus 30% ethanol extract treated.

• Journal of Life Science
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• v.25 no.4
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• pp.416-424
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• 2015

Pinosylvin is a stilbenoid found in the Pinus species. Pinosylvin at ~pM to ~nM concentrations induces cell proliferation, cell migration and anti-inflammatory activity in endothelial cells. However, it was recently reported that pinosylvin at high concentrations (50 to 100 μM) induces cell death in bovine aortic endothelial cells. In this study, we conducted a series of experiments to discover how pinosylvin at a high concentration (50 μM) induces endothelial cell death. Pinosylvin at the high concentration was shown to induce endothelial cell apoptosis through enhancing caspase-3 activity, flip-flop of phosphatidyl serine, and nuclear fragmentation. We found that pinosylvin at the high concentration additively increased caspase-3 activity enhanced by serum-starvation or treatment with 100 μM etoposide. We also determined that pinosylvin at the high concentration promoted activations of c-Jun N-terminal kinase (JNK) and endothelial nitric oxide synthetase (eNOS). We further ran a series of experiments to find out which signaling molecule plays a critical role in the pinosylvin-induced apoptosis. We finally found that SP-600125, a JNK inhibitor, had an inhibitory effect on the pinosylvin-induced endothelial cell death, but L-NAME, an eNOS inhibitor, had no effect. These data indicate that JNK is involved in the pinosylvin-induced apoptosis. Collectively, pinosylvin at high doses induces cell apoptosis via JNK activation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.2
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• pp.124-130
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• 2007

In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOSI), alone or in combination, on the proliferation of cultured primary normal human oral keratinocytes (NHOK). The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. The DNA synthesis was measured by the BrdU assay. Addition of low concentration of AA ($1{\mu}M$) and high concentration of AA with NMDA group (NMDA+AA $10{\mu}M$) made DNA synthesis rate increase significantly at the early stage. Adding NNA ($10{\mu}M$) affected DNA synthesis rate to increase significantly in 4 hours. At the early stage, DNA synthesis was significantly active in the NOS-I with NMDA groups than in the control and the NMDA-only group, while it didn't become statistically meaningful in 24 hours. AA $1{\mu}M$ and NNA $10{\mu}M$ may induce the proliferation of the NHOK independently and NOS-I may induce the proliferation of the NHOK with NMDA. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.