• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.795-801
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• 2013

The purpose of this study is to investigate effects of White Ginseng-Ejung-tang (EG) and Red Ginseng-Ejung-tang (ER) water extract on production of various cytokines such as interleukin (IL)-21, IL-25, IL-$28{\beta}$, erythropoietin (EPO), Exodus-2, monocyte chemotactic protein (MCP)-5, macrophage inflammatory protein (MIP)-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, and TARC in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. ER significantly decreased levels of IL-21, IL-25, IL-$28{\beta}$, EPO, Exodus-2, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, TARC, and fractalkine for 24 h incubation at the oncentrations of 25 and 100 ${\mu}g/mL$ in LPS-induced RAW 264.7 (P < 0.05). But EG did not show any significant effect. These results suggest that ER has anti-inflammtory property related with its inhibition on the production of IL-21, IL-25, IL-$28{\beta}$, and chemokines such as EPO, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, Exodus-2, and TARC in LPS-induced macrophages.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1668-1674
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• 2016

Red ginseng, a steamed and sun-dried ginseng, is a popular health-promoting food in Korea and other Asian countries. We introduced nanofertilizer technology using gold nanoparticles in an effort to develop red ginseng with an elevated level of ginsenosides, the main active compounds of ginseng. Shoots of 6-year-old ginseng plants were fertilized three times with colloidal gold nanoparticle sprays. Red ginseng extract was prepared from the main roots. The concentrations of gold and ginsenosides were measured following gold nanoparticle treatment. To evaluate the anti-inflammatory effects, mouse peritoneal macrophages of male BALB/c mouse were stimulated with lipopolysaccharide plus interferon-γ in the presence of extracts from red ginseng with or without gold nanoparticle treatment. The content of ginsenosides, such as Rg1, Re, Rf, and Rb1, increased in ginseng treated with gold nanofertilizer whereas the steaming process increased only the levels of Rd and Rg3. The levels of nitric oxide, inducible nitric oxide synthase, and interleukin-6, but not tumor necrosis factor-α, were more suppressed in macrophages treated with extract from gold nanoparticle-treated red ginseng. Our results show that the use of a colloidal gold nanoparticle fertilizer improved the synthesis of ginsenosides in ginseng and enhanced the anti-inflammatory effects of red ginseng. Further research is required to elucidate the causal factors for the gold-induced change in ginsenoside synthesis and to determine the in vivo effect of gold nanoparticle-treated ginseng.

• Applied Microscopy
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• v.23 no.1
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• pp.91-108
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• 1993

The relationship of cartilage canals to initial osteogenesis of primary ossification center of developing vertebrae in human fetuses ranging from 50mm to 260mm in crown rump length was studied by light and electron microscopy. The cartiage canals of the thoracic vertebrae were first observed at 60mm fetus. Cartilage canals were identified as vascular channels arising from perichondrium surfaces. A number of cartilage canals were observed around the primary center of ossification at 80mm fetus. At 120mm fetus, cartilage canals of the bodies of vertebra were increased. Eventually the canals were eroded from the main medullary cavity and remained at only peripheral regions of growth cartilage. Superficial, intermediate, and deep canals were identified by the characteristics of cartilage cells. Fibroblasts, undifferentiated mesenchymal cells, and vacuolated macrophages were observed adjacent to the matrix of resting cartilage cells in the superficial canal. Fibroblasts and mesenchymal cells were densely packed at the tip of canal, giving an epithelial appearance to the clustered cell in the intermediate canal. Vacuolated macrophages were in contact with matrix of hypertrophied cartilage. The thick-walled vessels in the intermediate and deep canals consisted of typical endothelial cells, but in the newly formed vessels contained mesenchymal cells and fibroblasts incorporated into the vessel wall. During lengthening of cartilage canal, the matrix of cartilage cells were invaded by newly formed capillaries and vacuolated macrophages. At the deep canal, the lateral wall of the canal terminated in matrix containing calcified cartilage. The mesenchymal cells began to differentiate into osteoblasts adjacent to the calcified matrix. The results indicate that the connective tissue cells within the cartilage canals proliferate and differentiate into osteoblasts at the site of primary ossification center.

• Environmental Analysis Health and Toxicology
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• v.15 no.3
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• pp.69-80
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• 2000

ADP-rubosylation may be involved in the process of macrophage activation. Nitric oxide (NO) has emerged as an important intracellular and interacellular regulatory molecule with function as diverse as vasodilation, neural communication or host defense. NO is derived from the oxidation of the terminal guanidino nitrogen atom of L-arginine by the NADPH -dependent enzyme, nitric oxide synthase (NOS) which is one of the three different isomers in mammalian tissues. Since NO can exert protective or regulatory functions in the cell at a low concentration while toxic effects at higher concentrations, its role may be tightly regulated in the cell. Therefore, this paper was focused on signal transduction pathway of NO synthesis, role of endogenous TGF-$\beta$ in NO production. effect of NO on superoxide formation. Costimulation of murine peritoneal macrophages with interferon-gamma (IFN-γ) and phorbol 12-myristate 13-acetate (PMA) increased both NO secretion and mRNA expression of inducible nitric oxide synthase (iNOS) when PMA abolished costimulation. Pretreatmnet of the cells with PMA abolished costimuation effects due to the depletion of protein kinase C (PKC) activities . The involvement of PKC in NO secretion could be further confirmed by PKC inhibitor, stauroprine, and phorbol ester derivative, phorbol 12,13-didecanoate. Addition of actinomycine D in IFN-γ plus PMA stimulated cells inhibited both NO secretion and mRNA expression of iNOS indication that PMA stabilizes mRNA of iNOS . Exogenous TGF-$\beta$ reduced NO secretion in IFN -γ stimulated murine macrophages. However addition of antisense oligodeoxynucleotide (ODN) to TGF-$\beta$ to this system recovered the ability of NO production and inhibited mRNA expression of TGF-$\beta$. ACAS interactive laser cytometry analysis showed that transportation of FITC -labeled antisense ODN complementary to TGF-$\beta$ mRNA could be observed within 5 min and reached maximal intensity in 30 min in the murine macrophage cells. NO released by activated macrophages inhibits superoxide formation in the same cells . This inhibition nay be related on NO-induced auto -adenosine diphosphate (ADP) -ribosylation . In addition, ADP-ribosylation may be involved in the process of macrophage activation .

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.2
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• pp.201-208
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• 1997

It is becoming increasingly clear that the inflammatory reaction can be ascribed to a complex array of mediators generated and released from activated phagocytes. In this study, the effect of PAF on interleukin-1(IL-1) activity by rat alveolar macrophages(AM) was examined using thymocyte proliferation assay in the supernate of sample obtained after 24 hr culture. When AM were cultured with PAF alone, no change in IL-1 activity was observed. However, the combined addition of PAF and muramyl dipeptide(MDP) or lipopolysaccharide(LPS) to AM cultures markedly enhanced IL-1 activity by 2-3 fold compared with AM cultures with the stimulant alone in a concentration dependent fashion. The peack effect was found at $10^{-8}$ M PAF with MDP and $10^{-14}$ M PAF with LPS. the effect of PAF was also tested in silica, toxic respirable dust, -added AM cultures as well as in the cultures containing bacterial compounds. Although silica did not stimulate the IL-1 activity, PAF could enhance IL-1 activity by 2 fold above the value of the silica-treated AM cultures with the peak response at $10^{-12}$ M PAF. Optimal enhancement of IL-1 activity occured when MDP and PAF were present together at the initiation of the 24 hr AM cultures. Additionaly, the biologically inactive precursor/metabolite of PAF, lyso-PAF failed to induce enhancement of IL-1 activity. When the specific, but structurally different PAF receptor antagonists, BN 52021($10^{-5}$ M) and CV 3988($10^{-5}$ M) was treated 15 min before addition of PAF($10^{-8}$ M) and MDP$(10\;{\mu}g/ml)$ to the AM cultures, it markedly inhibited the enhancement of IL-1 activity induced by PAF. The effects of these PAF antagonists were also observed in LPS$(10\;{\mu}g/ml)$-stimulated cells. Collectively, these data suggest that PAF enhances IL-1 activity by interaction with a specific receptor.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1047-1055
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• 2015

For the search of a potent first-in-class compound to inactivate macrophages responsible for inflammatory responses, in the present study, we investigated the anti-nflammatory effects of YH-1118, a novel synthetic compound, in a lipopolysaccharide (LPS)-stimulated mouse macrophage cell line, Raw 264.7. YH-1118 inhibited LPS-induced nitric oxide (NO) production and inducible NO synthase (iNOS) expression at both the protein and mRNA levels. The suppression of LPS-induced iNOS expression by YH-1118 was mediated via nuclear factor kappa B (NF-κB), but not activator protein-1 (AP-1) transcription factor. This was supported by the finding that YH-1118 attenuated the phosphorylation of inhibitor of κBα (IκBα) and nuclear translocation and DNA binding activity of NF-κB. Through the mechanisms that YH-1118 inhibited the activation of IκB kinases (IKKs), upstream activators of NF-κB, or p38 MAPK, YH-1118 significantly suppressed LPS-induced production of pro-inflammatory cytokines, tumor necrosis factor-α, interleukin-1β (IL-1β), and IL-6 (p < 0.05). In conclusion, our results suggest that YH-1118 inhibits LPS-induced inflammatory responses by blocking IKK and NF-κB activation in macrophages, and may be a therapeutic candidate for the treatment of various inflammatory diseases.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.4
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• pp.325-331
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• 2000

Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ plays important roles in inflammatory responses. Some of tetrahydroisoquinoline (THI) compounds exhibited to inhibit iNOS expression in animal studies and RAW 264.7 cells, but the action of THI on inflammatory reaction was not fully investigated. In the present study, we examined a limited series of THIs (higenamine, YS-51 and THI-52) on the $TNF-{\alpha}$ mRNA expression in mouse peritoneal macrophages by Northern analysis. When thioglycollate-stimulated peritoneal macrophages were incubated with LPS (100 ng/ml), expression of $TNF-{\alpha}$ mRNA was evident and reached its maximum at 2.5 h, which was reduced concentration-dependently by treatment with THIs. When the $TNF-{\alpha}$ activity of macrophage-conditioned media was measured using a TNF-sensitive L929 fibroblast cell line, CCL 1, all THIs increased the cell viability in a concentration dependent manner. The concentrations of THIs used are not cytotoxic by itself when analysed by MTT. Furthermore, nitrite/nitrate level was significantly reduced by the presence of THIs in cells treated with $LPS+interferon-{\gamma}\;(IFN-{\gamma}).$ It is concluded, thus, that these results strongly indicated that THIs can suppress the $TNF-{\alpha}$ expression and reduce NO, which may be useful for the inflammatory disorders.

• Nutritional Sciences
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• v.5 no.3
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• pp.123-128
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• 2002

Physical activity is a primary cancer control strategy that has received little attention to date. However, an Increasing number of epidemiological studies have proposed that physical exercise may be beneficial by enhancing anticancer immune system responses. We investigated the effects of acute exercise on changes in nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression. The amounts of NO generated by abdominal macrophages in mice were measured after exercise. Thirty-two mice, which were challenged with thioglycollate broth to activate peritoneal macrophages, were randomly assigned to control, exercise and recovery groups. The mice exercised on a motor-driven treadmill for 3 consecutive days, either moderately (18m/min, 30 min/day, 5％ grade) or severely (18-35m/min, 60 min/day, 5％ grade). The mice were killed immediately after exercise or after 6 hrs of recovery. Nitric oxide was quantified by the Griess assay. The exercised mice showed higher levels of NO generation than those of the control mice, but the intensity of exercise had no significant effect on NO generation. Mice allowed six hours of recovery after exercise showed higher levels of NO generation than that of animals sacrificed immediately after exercise, but there were no significant differences in NO generation with variations in the intensity of exercise. Increased levels of iNOS were found in the exercised groups, and this was greatest in the groups allowed six hours of recovery compared to those groups sacrificed immediately after exercise. The results of this study suggest that acute exercise may enhance an immune response by inducing macrophage-derived NO generation; these results support the epidemiological findings which support the benefits of exercise in the prevention and control of cancer. Further study is needed to determine the physiological significance of these findings, which could be applied to the use of therapeutic exercises to assist in the prevention and control of cancer.

• Korean Journal of Medicinal Crop Science
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• v.16 no.5
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• pp.313-319
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• 2008

Scutellaria barbata was examined to evaluate its modulatory effects on the functional activation of macrophages under lipopolysaccharide (LPS) treatment. To do this, hot water extract (Sb-HWE) was prepared from Scutellaria barbata and several inflammatory parameters such as nitric oxide (NO) production, phagocytosis, reactive oxygen species (ROS) determination and intracellular signaling pathway were selected to be tested. Sb-HWE strongly blocked NO production in LPS-activated RAW264.7 cells in a dose-dependent manner. However, it did not suppress inducible NO synthase (iNOS). In agreement, Sb-HWE did not diminish inflammatory signaling composed of NF-${\kappa}B$ and its upstream activation signaling enzymes such as Akt and $I{\kappa}B{\alpha}$. Sb-HWE protected RAW264.7 cells from LPS-induced cytotoxicity up to 80% at 400\;{\mu}g/ml$. Furthermore, this extract blocked phagocytic uptake of FITC-dextran, while sodium nitroprusside (SNP)-induced ROS generation in RAW264.7 cells was not decreased. Therefore, our data suggest that Sb-HWE may have differential immunoregulatory function depending on macrophage-mediated immune responses.

• Journal of Acupuncture Research
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• v.29 no.1
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• pp.67-74
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• 2012

Objectives : The objective of this study is to study the effects of hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ on nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production in macrophage. Methods : $Lonicera$ $japonica$ $Flos$ was extracted in two ways. One was extracted with distilled water(2L) for 4 h and the other one was extracted with 70% ethanol (2L) for 4h. The RAW 264.7 macrophage was subclutured. In order to evaluate cytotoxicity, MTT assay was performed. The concentrations of NO were measured by Griess assay. The concentrations of $PGE_2$ were measured by enzyme immunoassay. Results : 25, $125{\mu}g/m{\ell}$ hot aqueous extract from $Lonicera$ $japonica$ $Flos$ inhibited NO production in LPS-stimulated RAW 264.7 macrophages significantly. 25, 125, $625{\mu}g/m{\ell}$ ethanol extract from $Lonicera$ $japonica$ $Flos$ inhibited NO production in LPS-stimulated RAW 264.7 macrophages significantly. 150, $200{\mu}g/m{\ell}$ hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ inhibited $PGE_2$production in LPS-stimulated RAW 264.7 macrophages significantly. Conclusions : This study suggests that hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ suppress NO and $PGE_2$ production. So hot aqueous extract and ethanol extract from $Lonicera$ $japonica$ $Flos$ may have an anti-inflammation effect.