• The Korean Journal of Physiology and Pharmacology
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• v.17 no.2
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• pp.133-137
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• 2013

Vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), P- and E-selectin play a pivotal role for initiation of atherosclerosis. Ginsenoside, a class of steroid glycosides, is abundant in Panax ginseng root, which has been used for prevention of illness in Korea. In this study, we investigated the mechanism(s) by which ginsenoside Rg2 may inhibit VCAM-1 and ICAM-1 expressions stimulated with lipopolysaccharide (LPS) in human umbilical vein endothelial cell (HUVEC). LPS increased VCAM-1 and ICAM-1 expression. Ginsenoside Rg2 prevented LPS-mediated increase of VCAM-1 and ICAM-1 expression. On the other hand, JSH, a nuclear factor kappa B (NF-${\kappa}B$) inhibitor, reduced both VCAM-1 and ICAM-1 expression stimulated with LPS. SB202190, inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), and wortmannin, phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, reduced LPS-mediated VCAM-1 but not ICAM-1 expression. PD98059, inhibitor of mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) did not affect VCAM-1 and ICAM-1 expression stimulated with LPS. SP600125, inhibitor of c-Jun N-terminal kinase (JNK), reduced LPS-mediated ICAM-1 but not VCAM-1 expression. LPS reduced IkappaB${\alpha}$ ($I{\kappa}B{\alpha}$) expression, in a time-dependent manner within 1 hr. Ginsenoside Rg2 prevented the decrease of $I{\kappa}B{\alpha}$ expression stimulated with LPS. Moreover, ginsenoside Rg2 reduced LPS-mediated THP-1 monocyte adhesion to HUVEC, in a concentration-dependent manner. These data provide a novel mechanism where the ginsenoside Rg2 may provide direct vascular benefits with inhibition of leukocyte adhesion into vascular wall thereby providing protection against vascular inflammatory disease.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.313-320
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• 2014

The study was conducted to investigate the role of vitamin E in the high altitude hypoxia-induced damage to the intestinal barrier in rats. Sprague-Dawley rats were divided into control (Control), high altitude hypoxia (HH), and high altitude hypoxia + vitamin E (250 mg/kg $BW^*d$) (HV) groups. After the third day, the HH and HV groups were placed in a hypobaric chamber at a stimulated elevation of 7000 m for 5 days. The rats in the HV group were given vitamin E by gavage daily for 8 days. The other rats were given equal volume saline. The results showed that high altitude hypoxia caused the enlargement of heart, liver, lung and kidney, and intestinal villi damage. Supplementation with vitamin E significantly alleviated hypoxia-caused damage to the main organs including intestine, increased the serum superoxide dismutase (SOD) (p< 0.05), diamino oxidase (DAO) (p< 0.01) levels, and decreased the serum levels of interleukin-2 (IL-2) (p< 0.01), interleukin-4 (IL-4) (p<0.001), interferon-gamma ($IFN-{\gamma}$) (p<0.01) and malondialdehyde (MDA) (p<0.001), and decreased the serum erythropoietin (EPO) activity (p<0.05). Administration of vitamin E significantly increased the S-IgA (p<0.001) in ileum and significantly improved the expression levels of occludin and $I{\kappa}B{\alpha}$, and decreased the expression levels of hypoxia-inducible factor 1 alpha and 2 alpha ($HIF-1{\alpha}$ and $HIF-2{\alpha}$), Toll-like receptors (TLR4), P-$I{\kappa}B{\alpha}$ and nuclear factor-${\kappa}B$ p65(NF-${\kappa}B$ P65) in ileum compared to the HH group. This study suggested that vitamin E protectis from intestinal injury caused by high altitude hypoxia environment. These effects may be related to the HIF and TLR4/NF-${\kappa}B$ signaling pathway.

• Korean Journal of Food Science and Technology
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• v.49 no.4
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• pp.453-458
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• 2017

This study compared the immuno-modulatory effects of ethanol extracts (A. muricata L. ethanol extracts, ALE) and crude polysaccharide fraction (A. muricata L. crude polysaccharide fraction, ALP) from Annona muricata L. in macrophages. Immuno-modulatory activity was determined by assessing cell viability, nitric oxide (NO) production, inducible NO synthase (iNOS) expression and cytokine production in RAW 264.7 a macrophage cell line. Both ALE and ALP treatment did not affect cytotoxicity, and ALP treatment significantly increased NO production. Additionally, cytokine production [tumor necrosis factor ($TNF-{\alpha}$; $2909.04{\pm}4.1pg/mL$), interleukin (IL)-6; $662.84{\pm}5.3pg/mL$, and $IL-1{\beta}$; $852.37{\pm}2.2pg/mL$), was highly increased in the ALP ($250{\mu}g/mL$) treated group compared to the ALE ($250{\mu}g/mL$) treated group ($TNF-{\alpha}$; $1564.50{\pm}6.1pg/mL$, IL-6; $517.24{\pm}4.1pg/mL$ and $IL-1{\beta}$; $237.23{\pm}1.8pg/mL$). Moreover, ALP treatment considerably increased the expression of mitogen-activated protein kinase (MAPKs) and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) in the macrophages. Therefore, ALP can induce macrophage activation through MAPK and $NF-{\kappa}B$ signaling and this can be a potential candidate for development of nutraceuticals.

• Biomolecules & Therapeutics
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• v.25 no.6
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• pp.641-647
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• 2017

Galangin (3,5,7-trihydroxyflavone) is a polyphenolic compound abundant in honey and medicinal herbs, such as Alpinia officinarum. In this study, we investigated the anti-inflammatory effects of galangin under in vitro and in vivo neuroinflammatory conditions caused by polyinosinic-polycytidylic acid (poly(I:C)), a viral mimic dsRNA analog. Galangin suppressed the production of nitric oxide, reactive oxygen species, and pro-inflammatory cytokines in poly(I:C)-stimulated BV2 microglia. On the other hand, galangin enhanced anti-inflammatory interleukin (IL)-10 production. Galangin also suppressed the expression of pro-inflammatory markers in poly(I:C)-injected mouse brains. Further mechanistic studies showed that galangin inhibited poly(I:C)-induced nuclear factor (NF)-${\kappa}B$ activity and phosphorylation of Akt without affecting MAP kinases. Interestingly, galangin increased the expression and transcriptional activity of peroxisome proliferator-activated receptor (PPAR)-${\gamma}$, known to play an anti-inflammatory role. To investigate whether PPAR-${\gamma}$ is involved in the anti-inflammatory function of galangin, BV2 cells were pre-treated with PPAR-${\gamma}$ antagonist before treatment of galangin. We found that PPAR-${\gamma}$ antagonist significantly blocked galangin-mediated upregulation of IL-10 and attenuated the inhibition of tumor necrosis factor (TNF)-${\alpha}$ and IL-6 in poly(I:C)-stimulated microglia. In conclusion, our data suggest that PI3K/Akt, NF-${\kappa}B$, and PPAR-${\gamma}$ play a pivotal role in mediating the anti-inflammatory effects of galangin in poly(I:C)-stimulated microglia.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.6
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• pp.1284-1290
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• 2002

Scrophularia buergeriana Miquel (Scrophulariaceae) has been used as an anti-inflammatory drug in the folk medicine recipe and been proved its anti-inflammatory effect in the oriental medicine. Since nitric oxide (NO) and superoxide anion (O/sub 2//sup -/) are ones of the major inflammatory parameters, we studied the effect of aqueous extracts of Scrophularia buergeriana (SB) on NO and O/sub 2//sup -/ production in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. NO, O/sub 2//sup -/production and inducible NO synthase (iNOS) level were significantly reduced in LPS-activated macrophages by SB compared to those without. Electrophoretic mobility shift assay (EMSA) indicated that SB blocked the activation of NF-kB, which was considered to be a potential transcription factor for the iNOS expression. SB also blocked degradation of IkBα. Furthermore, IkB kinase alpha (IKKα), which phosphorylates serine residues of IkB directly, is inhibited by SB. These results suggest that SB could exert its anti-inflammatory actions by suppressing the activation of NF-kB through inhibition of IKK activity.

• Korean Journal of Food Science and Technology
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• v.50 no.5
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• pp.555-563
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• 2018

Barley has nutritional benefits due to its high dietary fiber content; therefore, the intake of whole barley grains is recommended. However, barley is often consumed in the fermented form because of the improved texture and digestibility. The present study was designed to elucidate the intracellular signaling pathway for macrophage activation by the polysaccharide BF-CP from fermented barley. BF-CP is a neutral polysaccharide, composed of neutral sugars, including glucose (70.7%), xylose (11.4%), and arabinose (9.0%). BF-CP exhibited macrophage-stimulatory activity by inducing the production of interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide in RAW 264.7 macrophages. Further, BF-CP treatment strongly increased the IL-6 and $TNF-{\alpha}$ gene expression in a concentration-dependent manner. Signal transduction experiments using immunoblotting showed that BF-CP phosphorylated mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38, and nuclear factor $(NF)-{\kappa}B$, in RAW 264.7 cells in a concentration-dependent manner. These results suggest that BF-CP activates the macrophages via MAPK and $NF-{\kappa}B$ pathways, and also induces an increase in the production of cytokines.

• IMMUNE NETWORK
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• v.10 no.2
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• pp.55-63
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• 2010

Background: Cordyceps militaris has been used in traditional medicine to treat numerous diseases and has been reported to possess both antitumor and immunomodulatory activities in vitro and in vivo. However, the pharmacological and biochemical mechanisms of Cordyceps militaris extract (CME) on macrophages have not been clearly elucidated. In the present study, we examined how CME induces the production of proinflammatory cytokines, transcription factor, and the expression of co-stimulatory molecules. Methods: We confirmed the mRNA and protein levels of proinflammatory cytokines through RT-PCR and western blot analysis, followed by a FACS analysis for surface molecules. Results: CME dose dependently increased the production of NO and proinflammatory cytokines such as IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and $PGE_2$, and it induced the protein levels of iNOS, COX-2, and proinflammatory cytokines in a concentrationdependent manner, as determined by western blot and RT-PCR analysis, respectively. The expression of co-stimulatory molecules such as ICAM-1, B7-1, and B7-2 was also enhanced by CME. Furthermore, the activation of the nuclear transcription factor, NF-${\kappa}B$ in macrophages was stimulated by CME. Conclusion: Based on these observations, CME increased proinflammatory cytokines through the activation of NF-${\kappa}B$, further suggesting that CME may prove useful as an immune-enhancing agent in the treatment of immunological disease.

• Journal of Life Science
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• v.27 no.5
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• pp.509-516
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• 2017

Perilla frutescens (L.) Britton var. sprouts (PFS) is a plant of the labiatae family. The purpose of this work was to assess the preventive effects of PFS ethanolic extracts (PFSEs) on cytokine-induced ${\beta}$-cell damage. Cytokines, which are released by the infiltration of inflammatory cells around the pancreatic islets, are involved in the pathogenesis of type 1 diabetes mellitus. The combination of interleukin-$1{\beta}$ (IL-1), interferon-${\gamma}$ (IFN-${\gamma}$), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) induced formation of reactive oxygen species (ROS). Accumulation of intracellular ROS led to ${\beta}$-cell dysfunction and apoptosis. PFSEs possess antioxidant activity and thus lead to downregulation of ROS generation. Cytokines decrease cell viability, stimulate the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and induce the production of nitric oxide (NO). PFSEs prevented cytokine-induced cell viability in a dose-dependent manner. Incubation with PFSE resulted in significant reduction in cytokine-induced NO production that correlated with reduced levels of the iNOS and COX-2 protein expression. Furthermore, PFSE significantly decreased the activation of nuclear factor ${\kappa}B$ (NF-${\kappa}B$) by inhibition of $I{\kappa}B{\alpha}$ phosphorylation in RINm5F cells. In summary, our results suggest that the protective effects of PFSE might serve to counteract cytokine-induced ${\beta}$-cell destruction. Findings indicate that consumption of Perilla frutescens (L.) Britton var. sprouts alleviates hyperglycemia-mediated oxidative stress and pro-inflammatory cytokine-induced ${\beta}$-cell damage and thus has beneficial anti-diabetic effects.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.5
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• pp.527-536
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• 2014

An ethanolic extract from Myagropsis yendoi was fractionated using several solvents. Among these, an ethyl acetate fraction (Myagropsis yendoi ethyl acetate fraction: MYE) showed the highest anti-inflammatory activity based on inhibition of lipopolysaccharides (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells. We thus investigated the molecular mechanisms underlying MYE's inhibitory effects. Pretreatment of cells with up to $30{\mu}g/mL$ of MYE significantly inhibited NO production and inducible nitric oxide synthase expression in a dose-dependent manner (P<0.05). Similarly, MYE markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor (TNF)-${\alpha}$, as well as their mRNA levels. While the nuclear translocation of nuclear factor-kappa B (NF-${\kappa}B$) was strongly suppressed by MYE, the activation of a nuclear factor erythroid 2-related factor (Nrf2) was increased. Moreover, MYE significantly reduced the phosphorylation of JNK, p38 MAPK, and phosphatidylinositol 3-kinase/Akt in LPS-stimulated cells. These results indicate that MYE contains anti-inflammatory compounds, and that it might be used as a dietary supplement for the prevention of inflammatory diseases.

• Archives of Pharmacal Research
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• v.25 no.5
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• pp.691-696
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• 2002

Doxorubicin (DOX) is an active and broad spectrum chemotherapeutic agent. Increased inducible nitric oxide synthase (NOS) expression and/or activity have been reported in several human tumors. While the relationship between DOX treatment and the enzymatic activity of endothelial NOS has been well characterized, little is known about the effects of DOX on the expression of iNOS in human cancer cells. In the present study, we characterized the effects of DOX on the nitric oxide (NO) production by colorectal cancer cells, DLD-1. IFN-${\gamma}$/IL-1$\beta$ (CM) increased the production of NO, whereas pretreatment of DOX inhibited the production of NO in response to CM in a dose dependent manner. The increased expressions of iNOS mRNA and protein by CM were completely blocked by DOX without affecting the iNOS mRNA stability. However, DOX activated nuclear factor-kB (NF-kB) in response to CM. Furthermore, the expression of inhibitor kB$\alpha$ was reduced by DOX in a dose dependent manner. Collectively, DOX inhibited the production of NO by DLD-1 cells, which is not linked to well known transcription factor, NF-kB. Therefore, further studies on the possible mechanisms of inhibitory effects of NO production by DOX would be worth pursuing.