• Biomolecules & Therapeutics
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• v.32 no.1
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• pp.154-161
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• 2024

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive paralysis. L-Citrulline is a nonessential neutral amino acid produced by L-arginine via nitric oxide synthase (NOS). According to previous studies, the pathogenesis of ALS entails glutamate toxicity, oxidative stress, protein misfolding, and neurofilament disruption. In addition, L-citrulline prevents neuronal cell death in brain ischemia; therefore, we investigated the change in the transport of L-citrulline under various pathological conditions in a cell line model of ALS. We examined the uptake of [¹⁴C]L-citrulline in wild-type (hSOD1wt/WT) and mutant NSC-34/ SOD1^G93A (MT) cell lines. The cell viability was determined via MTT assay. A transport study was performed to determine the uptake of [¹⁴C]L-citrulline. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to determine the expression levels of rat large neutral amino acid transported 1 (rLAT1) in ALS cell lines. Nitric oxide (NO) assay was performed using Griess reagent. L-Citrulline had a restorative effect on glutamate induced cell death, and increased [¹⁴C]L-citrulline uptake and mRNA levels of the large neutral amino acid transporter (LAT1) in the glutamate-treated ALS disease model (MT). NO levels increased significantly when MT cells were pretreated with glutamate for 24 h and restored by co-treatment with L-citrulline. Co-treatment of MT cells with L-arginine, an NO donor, increased NO levels. NSC-34 cells exposed to high glucose conditions showed a significant increase in [¹⁴C]L-citrulline uptake and LAT1 mRNA expression levels, which were restored to normal levels upon co-treatment with unlabeled L-citrulline. In contrast, exposure of the MT cell line to tumor necrosis factor alpha, lipopolysaccharides, and hypertonic condition decreased the uptake significantly which was restored to the normal level by co-treating with unlabeled L-citrulline. L-Citrulline can restore NO levels and cellular uptake in ALS-affected cells with glutamate cytotoxicity, pro-inflammatory cytokines, or other pathological states, suggesting that L-citrulline supplementation in ALS may play a key role in providing neuroprotection.

• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.79-89
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• 2020

This study was performed to evaluate the anti-inflammatory activities of 70% ethanol extract from Ambrosia trifida L. (AT). The electron donating ability and ABTS⁺ radical scavenging ability of extract from AT was shown to be 84.1% and 92.5% at 1,000 ㎍/ml concentration. The astringent effect of extract from AT was shown to be 94.7% at 1,000 ㎍/ml. The anti- inflammatory activities of extract of AT were investigated using RAW 264.7 cells induced by lipopolysaccharide (LPS). The cell toxicity effect of AT extract on RAW 264.7 performed MTT assay. As a result of the measured cell toxicity effect, 90% or more was shown with cell viability at a 500 ㎍/ml concentration. In nitric oxide synthesis inhibition effect, it was shown that extract from AT concentration dependent inhibited nitric oxide production. The protein expression inhibitory effect of AT extract was measured by western blot at 25, 50, and 100 ㎍/ml concentration and the β-actin used as a positive control. Consequently, the inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 protein expression inhibitory effect was decreased by 8.6%, 25.1% at 100 ㎍/ml concentration. The phosphorylation of extracellular signal-regulated kinase 1/2, p38, c-Jun NH₂-terminal kinase and Iκ-Bα protein expression inhibitory effect was a decreased dependent concentration. The mRNA expression inhibitory effect was measured by reverse transcription - polymerase chain reaction at 25, 50, and 100 ㎍/ml concentration and the glyceraldehyde-3-phosphate dehydrogenase used as a positive control. Consequently, the iNOS, COX-2, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α mRNA expression inhibition effect was a decreased dependent concentration in an LPS-activated macrophage. In conclusion, AT extract may have some effects on inflammatory factors as potential anti-inflammatory agents and natural substance for cosmetics.

• Journal of Life Science
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• v.26 no.3
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• pp.318-324
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• 2016

This study was aimed to verify anti-inflammatory activity of fermented Sargassum siliquanstrum with lactic acid bacteria. Anti-inflammatory activities were compared by measuring the amount of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and suppressive effect on inducible nitric oxide synthase (iNOS) expression in stably transfected RAW 264.7 cells. Inhibitory activities of NO production and iNOS expression were measured after confirmation of NO radical scavenging activities. Fermentation increased NO radical scavenging activities from 7.6% to 15.2% compared to non-fermented condition, and fermentation with Lactobacillus sp. SH-1 was the most efficient. Fermentation without algal debris showed better NO radical scavenging activities than that with debris. Fermentation with Lactobacillus sp. SH-1 also showed the highest NO production inhibitory activity (64.1%) in LPS-stimulated RAW 264.7 cells. LPS-induced iNOS expression was diminished to 28.6, 35.6, 49.4 and 58.5 at 50, 100, 500 and 1,000 μg/ml, respectively, by fermentation with Lactobacillus sp. SH-1. According to MTT assay, fermented S. siliquanstrum did not influence the cell viability at all concentrations tested, meaning no or less cytotoxicity. These results suggest that S. siliquanstrum has NO radical scavenging activity and anti-inflammatory activity. Thus biological activities of S. siliquanstrum were upgraded by fermentation, which could be used for the development of functional foods.

• Korean Journal of Plant Resources
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• v.26 no.5
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• pp.533-538
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• 2013

Potatoes were first introduced outside the Andes region four centuries ago, and have become an integral part of much of the world's food. Potatoes were first introduced into Europe in the 16th century and Korea in the early 19th century. In the nutritional aspects, potatoes contain abundant vitamins and minerals, as well as an assortment of phytochemicals such as carotenoids and natural phenols. Chlorogenic acid constitutes up to 90% of potato natural phenols. Due to the high content of potato functional compounds, it has known that potatoes are effective in the prevention of various human diseases. Recently, color-fleshed potatoes 'Hongyoung' and 'Jayoung' were developed by RDA, and it has reported that they have high content of anthocyanin. Additionally they show higher radical scavenging activity compared to white or yellow fleshed potatoes. So it will be expected that the consumption of color-fleshed potatoes grandually increase by pre-peeled potatoes and color potato chips. This study was conducted to enhance the peel of color-fleshed potatoes utilization and to determine the biological activity of peel of color-fleshed potatoes extract. The anti-inflammatory effects on ethanol extract and its solvent fraction were also evaluated. The anti-inflammatory activities of $CHCl_3$ fraction was evaluated for inhibitory activities against lipopolysacchride(LPS) induced nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production as well as inducible nitric oxide synthase(iNOS) and cyclo oxygenase-2(COX-2) protein expressions in RAW264.7 cell lines. The fraction inhibitory activity for both tests with $IC_{50}$ values showed in the ranges of $25{\sim}50{\mu}g/ml$. This result revealed that $CHCl_3$ fraction of Jayoung's peel is expected to be good candidate for development into source of anti-inflammatory agent.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2001.11a
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• pp.100-100
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• 2001

Hypoxia is a pathophysiological condition that occurs during injury, ischemia, and stroke. Hypoxic stress induces the expression of genes associated with increased energy flux, including the glucose transporters Glutl and Glut3, several glycolytic enzymes, nitric oxide synthase, erythropoietin and vascular endothelial growth factor. Induction of these genes is mediated by a common basic helix-loop-helix PAS transcription complex, the hypoxia-inducible factor-l${\alpha}$ (HIF-1${\alpha}$)/ aryl hydrocarbon receptor nuclear translocator (ARNT). Insulin plays a central role in regulating metabolic pathways associated with energy storage and utilization. It triggers the conversion of glucose into glycogen and triglycerides and inhibits gluconeogenesis. Insulin also induced hypoxia-induced genes. However the underlying mechanism is unestablished. Here, we study the possibility that transcription factor HIF-1${\alpha}$ is involved in insulin-induced gene expression. We investigate the mechanism that regulates hypoxia-inducible gene expression In response to insulin We demonstrate that insulin increases the transcription of hypoxia- inducible gene. Insulin-induced transcription is not detected in Arnt defective cell lines. Under hypoxic condition, HIF- l${\alpha}$ stabilizes but does not under insulin treatment. Insulin-induced gene expression is inhibited by presence of PI-3 kinase inhibitor and Akt dominant negative mutant, whereas hypoxia-induced gene expression is not. ROS inhibitor differently affects insulin-induced gene expressions and hypoxia-induced gene expressions. Our results demonstrate that insulin also regulates hypoxia-inducible gene expression and this process is dependent on Arnt. However we suggest HIF-l${\alpha}$ is not involved insulin-induced gene expression and insulin- and hypoxia- induces same target genes via different signaling pathway.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.117-125
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• 2007

Objectives : Berberine, a main alkaloid component of Coptidis rhizoma, has an antimicrobial and anti-tumor activities and antiinflammatory effect. In the present study, we investigated effect of berberine on the production of inflammatory mediators such as nitric oxide(NO), prostaglandin E2(PGE2), TNF-${\alpha}$ and IL-1${\beta}$ in LPS-stimulated BV2 microglial cells, Methods : BV2 cells were pre-treated with berberine and then stimulated with LPS. The cytotoxicity of berberine was determined by MTT assay. The NO production was measured by Griess assay. The mRNA expression and protein levels of inducible nirtic oxide synthase(iNOS) were determined by RT-PCR and Western blot. The production of PGE2 and cytokines was measured by ELISA. Results : Berberine inhibited the production of NO, PGE2 and pro inflammatory cytokines, TNF-${\alpha}$ and IL-1${\beta}$ in a dose dependent manner in LPS-stimulated BV2 cells. In addition, berebrine greatly suppressed the mRNA expression and protein levels of iNOS and inflammatory cytokines induced by LPS stimulation. These results indicate that the post-transcriptional regulatory mechanism of iNOS and/or inflammatory cytokine gene expression by berberine is involved in its anti-inflammatory effects, respectively. Conclusion : The present study suggests that berberine can be useful as a potential anti-inflammatory agent for treatment of various neurodegenerative diseases such as Alsheimer's disease, Parkinson's disease and stroke.

• Culinary science and hospitality research
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• v.18 no.4
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• pp.233-242
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• 2012

Macrophage-derived nitric oxide (NO) plays an important role in immune responses. Overproduction of NO by inducible nitric synthase(iNOS) is known to be closely correlated with the pathology of a variety of diseases and inflammations. This study investigated the inhibitory effect of extract from various mushrooms(Phellinus linteus(PL), Inonotus obliquus(IO), Sarcodon aspratus(SA), Hericium erinacium(HE), and Russula cutefracta(RC)) on NO production and its molecular mechanism in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Among the mushrooms, HE extracts exhibited the strongest NO-inhibition activity. It was also found that HE extract inhibited LPS-induced iNOS and COX-2 expression in RAW264.7 cells. The present results give the understanding of biological activities of mushrooms and encourage their application for supplements and cook.

• Advances in Traditional Medicine
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• v.5 no.4
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• pp.251-261
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• 2005

Rheumatoid arthritis (RA) is an autoimmune/inflammatory disorder with a complex genetic component. RA is characterized by chronic inflammation of the synovial membrane in the joint, which leads to the progressive destruction of articular cartilage, ligament and bone. Several cytokines such as tumor necrosis $factor-{\alpha}\;TNF-{\alpha}\;and\;interleukin-1{\beta}\;(IL-1{\beta})$ and interleukin-6 (IL-6) have been implicated in the pathological mechanisms of synovial tissue proliferation, joint destruction and programmed cell death in rheumatoid joint. Genome wide screening of subjects suffering from autoimmune diseases especially arthritis revealed linkage to inflammatory molecules like $TNF-{\alpha},\;IL-1{\beta}$ and IL-6, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappaB $(NF-{\kappa}B)$ and human leucocyte antigen/major histocompatibility complex (HLA/MHC) locus. The status of the pharmacological mechanism of herbal drugs in the light of genome wide screening results has been discussed to reinforce the therapeutic potential and the pharmacological basis of the herbal drugs.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.248.3-249
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• 2002

Nitric oxide (NO) has been shown to play an important role in the regulation of vascular tone. platelet function. neurotransmission. and immune function. NO is synthesized from the L-arginine by NO synthase (NOS). Three distinct isoforms of NOS have been identified: calcium/calmodulin-dependent endothelial (eNOS) and neuronal (nNOS) isoforms which are constitutive and produce small quantities of NO, and an inducible isoform (iNOS) which is markedly induced in response to lipopolysaccharide (LPS) or inflammatory cytokines. (omitted)

• Journal of Haehwa Medicine
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• v.22 no.1
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• pp.37-48
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• 2013

This study aims to summarize and makes a reference of anti-inflammatory activities of herbal medicines. In this process, this review collated papers of anti-inflammation-focused studies using herbal medicines in Oriental medical journals since 2003. Finally 221 papers were included and the type of materials, the type and effective classification of herbal medicines, the type of cells used in the experiments and the action and mechanisms of herbal medicine were analysed. The herbal medicines having the effects of decreasing fire and tonifying and nourishment were used the most. Most herbal medicines in this study can decrease proinflammatory cytokines, NO and prostaglandin 2 (PGE 2) production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by regulating of nuclear factor kappa B (NF-${\kappa}B$) and/or mitogen activated protein kinases (MAPKs).