• Journal of Microbiology and Biotechnology
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• v.30 no.3
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• pp.352-358
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• 2020

In this study we investigated the immune effects of oral administration of anionic macromolecules extracted from Codium fragile (CFAM) and red ginseng extract mixture on the peritoneal macrophage cells in immune-suppressed mice. Cyclophosphamide (CY) induces the immune-suppressed condition. CY-treated mice were orally fed with different concentrations of CFAM supplemented with red ginseng extract and the peritoneal macrophages collected. CY treatment significantly decreased the immune activities of peritoneal macrophages, compared to the normal mice. The administration of CFAM mixed with red ginseng extract significantly boosted the viability of macrophage cells and nitric oxide production of peritoneal macrophages. Further, the oral administration of CFAM mixed with red ginseng extract up-regulated the expression of iNOS, COX-2, and TLR-4 as well as cytokines such as IL-1β, IL-6, TNF-α, and IFN-γ more than the red ginseng-treated group. This study showed that CFAM enhanced the immune activity of red ginseng extract in the peritoneal macrophage cells of immune-suppressed mice. Furthermore, CFAM might be used as a co-stimulant of red ginseng extract through the regulation of macrophage cells for the enhancement of human health and immunity.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.2
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• pp.233-239
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• 1998

Stimulated alveolar macrophages and neutrophils produce nitric oxide, a free radical by an inducible nitric oxide synthase(iNOS), which reacts with superoxide anion to form peroxynitrite, a more highly reactive toxic species. The objectives of the present study were to evaluate acute inflammatory lung injury and to determine iNOS mRNA induction and nitric oxide production by rat broncho-alveolar lavage cells following intratracheal treatment of silica. After 4 h exposure to silica, differential counts of broncho-alveolar lavage cells and lactate dehydrogenase(LDH) activity as well as total protein in the broncho-alveolar lavage fluid were determined. Broncho-alveolar lavage cells were also assayed for iNOS mRNA and the productions of nitrite and nitrate measured in the cells cultured. Differential analysis of broncho-alveolar lavage cells showed that the number of alveolar macrophages slightly decreased following silica treatment; however, red blood cells, lymphocytes, and neutrophils significantly were increased by 9-, 14-, and 119-fold following silica treatment, respectively, compared with the saline control. It was also found significant increases in the LDH activity and total protein in the lavage fluid obtained from silica-treated rats, indicating silica-induced acute lung injury. Northern blot analysis demonstrated that the steady state levels of iNOS mRNA in broncho-alveolar lavage cells were increased following silica treatment. The productions of nitrite and nitrate in the cultured cells were significantly increased by 2-fold following silica treatment, respectively, which were attenuated by the NOS inhibitor $N{\omega}-nitro-L-arginine-methyl$ ester(L-NAME) and partially reversed by L-arginine. These findings suggest that nitric oxide production in alveolar macrophages and recruited neutrophils is increased in response to silica. Nitric oxide may contribute in part to acute inflammatory lung injury.

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.355-373
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• 1999

Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.

• BMB Reports
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• v.47 no.6
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• pp.318-323
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• 2014

We isolated the phenolic glucoside salicortin from a Populus euramericana bark extract, and examined its ability to suppress inflammatory responses as well as the molecular mechanisms underlying these abilities, using lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Salicortin inhibited iNOS expression and the subsequent production of NO in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Salicortin significantly suppressed LPS-induced signal cascades of NF-${\kappa}B$ activation, such as IKK activation, $I{\kappa}B{\alpha}$ phosphorylation and p65 phosphorylation in RAW 264.7 cells. In addition, salicortin inhibited the LPS-induced activation of JNK, but not ERK or p38 MAPK. Furthermore, salicortin significantly inhibited production of pro-inflammatory cytokines, such as TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in the LPS-stimulated RAW 264.7 cells. These findings suggest that salicortin may show its anti-inflammatory activity by suppressing the LPS-induced expression of pro-inflammatory mediators through inhibition of NF-${\kappa}B$ and JNK MAPK signaling cascades in macrophages.

• Journal of Physiology & Pathology in Korean Medicine
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• v.30 no.4
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• pp.266-273
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• 2016

Ohmae-hwan has been used to treat the symptoms of loose stools mixed with blood accompanied by abdominal pain due to heat dysentery in Korean medicine. However, its mechanism of action on inflammatory disease using macrophages was not known. The author was to find out about the influence of Ohmae-hwan water extract on expressions of NO, cytokine IL-6 and TNF-α in LPS-stimulated RAW 264.7 cell, on expressions of mRNA of iNOS, IL-6 and COX-2 and on suppression of expressions of protein of iNOS and COX-2, and was to find out about the mechanism. As a result of the study, it was found that Ohmae-hwan suppressed generation of NO activated by LPS depending on its concentrations, suppressed expressions of cytokine IL-6, and TNF-α and suppressed expressions of mRNA of iNOS, IL-6, and COX-2 and suppressed expressions of protein of iNOS, COX-2. Considering the above results, it can be known that Ohmae-hwan water extract has anti-inflammatory effect against inflammatory response mediated by macrophages by suppressing generation of NO, TNF-α, and IL-6 and expressions of iNOS, COX-2 as it acts on macrophages.

• Natural Product Sciences
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• v.16 no.4
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• pp.265-270
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• 2010

The methanol extract of Zanthoxylum rhetsa (MZRR) were evaluated for its ability to suppress the formation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. MZRR presented an inhibition of LPS-induced production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in RAW 264.7 macrophages. Western blotting and RT-PCR analyses demonstrated that MZRR significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages in a dose-dependent manner. LPS-induced COX-2, iNOS, and nuclear factor kappa beta (NF-${\kappa}B$) activity were also decreased in the presence of MZRR. The production of tumor necrosis factor-$\alpha$ (TNF-$\alpha$), the mRNA expression levels of pro-inflammatory cytokines, including TNF-$\alpha$ and IL-$1{\beta}$, were reduced after MZRR administration in a dose dependent-manner. These results suggest that the MZRR extract involved in the inhibition of iNOS and COX-2 via the NF-${\kappa}B$ pathway, revealing a partial molecular basis for anti-inflammatory properties of the MZRR extract.

• Biomolecules & Therapeutics
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• v.17 no.3
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• pp.276-281
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• 2009

Lactic acid bacteria (LAB) are considered as probiotics with immunostimulatory property. In this study, we investigated the molecular mechanism of its immunostimulating potency on macrophages using combined preparation of LAB (cpLAB). cpLAB is able to strongly stimulate nitric oxide (NO) production as well as inducible NO synthase (iNOS) expression from macrophage-like RAW264.7 cells. The cpLAB-induced NO release seemed to be mediated by extracellular signal-regulated kinase (ERK) but not p38 and C-Jun N-terminal kinase (JNK), since U0126, an ERK inhibitor, clearly suppressed NO production. cpLAB significantly diminished the binding of toll like receptor (TLR)-2 antibody up to 25%, implying that cpLAB-mediated activation of macrophages may be required for the functional activation of TLR-2, but not TLR-4. Therefore, our data suggest that cpLAB may directly allow macrophages to immunostimulating potency via activation of TLR-2 and ERK.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.1
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• pp.39-54
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• 2008

Purpose: The purpose of this study was to investigate the anti-inflammatory effects of the water extract of Omisodokeum (OMSDE) on peritoneal macrophages, Methods: To verify the anti-inflammatory mechanism of OMSDE, the activation of nuclear $factor-{\kappa}B$ $(NF-{\kappa}B)$ and the phosphorylation of MAPK were examined. Results: The extract of OMSDE suppressed the production of LPS-induced nitric oxide (NO), tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, IL-6 and IL-12 in the macrophages. OMSDE inhibited the degradation of inhibitory ${\kappa}B-{\alpha}$ $(I{\kappa}B-{\alpha})$ and it suppressed the activation of extracellular signal-regulated kinase (ERK 1/2) but didn't inhibit c-Jun N-terminal kinase (JNK) and p38, indicating that OMSDE may inhibit the pro-inflammatory cytokine production process by inhibiting the activation of $NF-{\kappa}B$ and ERK 1/2. Furthermore, OMSDE inhibited the production of interferon $(IFN)-{\beta}$ but didn't inhibit of $IFN-{\alpha}$ in the LPS-stimulated macrophages through the down-regulation of interferon regulatory factor (IRF)-1 and IRF-7. The Oral administration of OMSDE inhibited LPS-induced endotoxin shock and the production of $TNF-{\alpha}$ in serum but didn't inhibit of $IL-1{\beta}$ and IL-6. Conclusion: These results suggest that OMSDE may be effective in the prevention and treatment of inflammatory diseases.

• Nutritional Sciences
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• v.5 no.4
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• pp.203-210
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• 2002

Aster scaber $T_{HUNB}$ (AST ; Charm-chui), a potent herbal medicinal plant, has a long tradition of use, being harvested as a wild plant, is said to stimulate appetite, and may act as a diuretic, antifebrile agent and painkiller. This study was conducted to investigate the immunomodulative effects of AST In mice, using in vitro and in vivo experiments. The immunomodulative effects were studied in vitro by measuring the proliferation of mice splenocytes and the production of three kinds of cytokines (IL-$\beta$, IL-6, and TNF-$\alpha$) by mice peritoneal macrophages which were cultured with sequential fractions of AST methanol extract (methanol, hexane, chlo-roform, ethylacetate, butanol and water). In an in vivo experiment using mice, different concentrations of AST water extract were orally administrated every other day for two weeks. The production of cytokines (IL-1$\beta$, IL-6, and TNF-$\alpha$) secreted by activated macrophages, and the proliferation of mice splenocytes, were used as indices for immunocompetence. In vitro supplementation using six fractions of AST in the range of 1 to 100$\mu$ g/ml enhanced splenocyte proliferation by 10.5% to 53% compared to the control. IL-1$\beta$production was significantly increased with the supplementation of butanol and water extracts of AST. Higher levels of IL-6 and TNF-$\alpha$production were detected with supplementation of methanol, ethylacetate, butanol or water extracts at the concentration of 100$\mu$ g/ml. In the in vivo study, the highest proliferation of splenocytes was seen in the mice orally administrated with the AST water extract at the concentration of 500mg/kg body weight. In the case of cytokine production, there were no significant differences in the production of IL-1$\beta$and IL-6 among the treated groups and the control. However, TNF-$\alpha$released by activated peritoneal macrophages were augmented by the oral administration of AST water extract. These results indicate that AST may enhance the immune functions by regulating splenocyte proliferation and cytokine production capacity in mice.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.738-744
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• 2013

The purpose of this study is to investigate effects of White Ginseng-Ejung-tang water extract (EJ) on production of various cytokines such as interleukin (IL)-2, IL-5, IL-6, IL-10, IL-12p70, macrophage inflammatory protein (MIP)-2, vascular endothelial growth factor (VEGF), keratinocyte-derived chemokine(KC), tumor necrosis factor (TNF)-${\alpha}$, and granulocyte macrophage colony-stimulating factor (GM-CSF) 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. EJ significantly decreased levels of IL-2, IL-12p70, IL-5, MIP-2 for 24 h incubation at the concentrations of 25, 50, and 100 ${\mu}g/mL$ in LPS-induced RAW 264.7 (P < 0.05). EJ significantly decreased levels of IL-6 at the concentrations of 50 and 100 ${\mu}g/mL$ (P < 0.05). EJ significantly decreased levels of IL-10 and VEGF at the concentrations of 25 and 100 ${\mu}g/mL$ (P < 0.05). EJ significantly decreased levels of KC at the concentrations of 100 ${\mu}g/mL$ (P < 0.05). EJ did not show any significant effect on TNF-${\alpha}$ and GM-CSF production. These results suggest that EJ has anti-inflammtory property related with its inhibition of IL-2, IL-5, IL-6, IL-10, IL-12p70, MIP-2, VEGF, and KC production in LPS-induced macrophages.