• Journal of the Korean Applied Science and Technology
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• v.40 no.6
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• pp.1268-1277
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• 2023

In this study, to evaluate the possibility of utilizing Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) leaf fractions as anti-inflammatory functional materials, C. obtusa extract extracted with 99% ethanol (CO99EL) was fractionated with hexane (CO99EL-H), chloroform (CO99EL-C), ethyl acetate (CO99EL-E), butanol (CO99EL-B) and distilled water (CO99EL-W). The anti-inflammatory effects of each fraction was performed using lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophages. Cytotoxicity was highest in CO99EL-H and CO99EL-C and lowest in CO99EL-W. Interestingly, LPS-induced iNOS expression and NO production were significantly reduced by CO99EL-H and CO99EL-E, and COX-2 expression was significantly reduced by CO99EL-B and CO99EL-W. In addition, interleukin (IL)-1𝛽, an inflammatory cytokine increased by LPS, was significantly reduced by CO99EL-C, CO99EL-E, CO99EL-B and CO99EL-W, and IL-6 was significantly reduced by CO99EL-B and CO99EL-W. Therefore, the janus kinase (JAK)/signaling transducer and activator of transcription (STAT) signaling pathway activated by LPS was significantly reduced by CO99EL-H and CO99EL-C, and the mitogen-activated protein kinase (MAPK) signaling pathway was slightly reduced by CO99EL-H and CO99EL-C. However, nuclear factor (NF)-𝜅B activity was not reduced by any fractions. Based on the results of this study, it was confirmed that CO99EL fractions have different anti-inflammatory mechanisms depending on the solvent used for fractionation.

• Tuberculosis and Respiratory Diseases
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• v.47 no.2
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• pp.161-171
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• 1999

Background: Interleukin-12 (IL-12) can induce antitumor effects in vivo. This antitumor effect is associated with T cell infiltration but the effect of IL-12 on the steps of T cell migration into the tumor tissue has not been fully elucidated. This study focused on the effect of IL-12 on the tumor growth and the metastasis and on the expression of E-selectin, an adhesion molecule which is activated endothelial specific in its expression. In addition, we studied whether the expression of E-selectin is associated with the TNF-$\alpha$, a cytokine that its production is increased by IL-12 and has functions inducing a variety of adhesion molecules. Methods: Mice of C57BL/6 strain were injected with Lewis lung cancer cells followed by either IL-12, TNF-$\alpha$, or normal saline by intraperitoneal route. Twenty eight days after tumor cell inoculation, metastatic nodules of lung were enumerated and immunohistochemical staining of the subcutaneous tumors were performed with monoclonal antibodies to CD4, CD8, CD16, and E-selectin. In IL-12 treated mice, the subcutaneously implanted Lewis lung tumors were decreased in size and the metastases were also decreased in number compared to control mice. On tumor tissues, increased infiltration of CD4+, CD8+, and CD16+ cells were oberved in IL-12 treated mice compared to control mice. In control mice, E-selectin was absent on tumor vessels, but the expression of E-selectin was increased on tumor vessels of IL-12 treated mice. Administration of TNF-$\alpha$ increased not only the expression of E-selectin but also infiltrations of CD4+, CD8+, and CD16+ cells on tumor tissues. Conclusions: These results demonstrate that IL-12 inhibits tumor growth and metastases through infiltrations of inflammatory cells in mouse model of Lewis lung carcinoma and E-selectin may playa role in inflammatory cell recruitment on tumor tissue following IL-12 administration. Also, TNF-$\alpha$ may have a role as a mediator responsible for the IL-12 induced expression of E-selectin.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.328-338
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• 1994

Background: Effect of sulfur dioxide($SO_2$) exposure on airway is well known but little about the effect of $SO_2$ exposure on lung parenchyme. This study is to determine if short tenn exposure to $SO_2$ in concentration commonly found in industrialized environment cause potentially harmful effect on the lung parenchyme, and to evaluate the exposure time-response relationship between short tenn exposure to $SO_2$ and the inflammatory response in mouse lung. Method: 5ppm $SO_2$ gas was used and 48 mice were grouped into control(10), 30(9), 60(11), and 120 minute exposure(18) group. In each group, bronchoalveolar lavage(BAL) was done immediately after and at 1,2,3 days after exposure. Histological examination was performed in control and 120 minute exposure group. Results: 1) Cell response in bronchoalveolar lavage fluid. In 30 and 60 minute exposure group, compared to the control group, lymphocyte count has significantly increased(p<0.05) at 1 day after exposure but did not differ at 2 days after exposure. In 120 minute exposure group, also compared to the control group, there was significant increase in total cell, macrophage, and lymphocyte count at 1 day after exposure, (p<0.05) which lasted for 2 days but did not significantly differ at 3 days after exposure. 2) Histological findings in 120 minute exposure group. In the airway, mild epithelial cell damage and ciliary loss were noted but there was no evidence of inflammatory cell infiltration. Interstitial inflammatory infiltration was noted at 1 day after exposure, which lasted for 3 days after exposure and there was no evidence of edema or fibrosis in the interstitium Conclusions: These data indicate potentially noxious effect of $SO_2$ on the lung parenchyme as well as the airway at exposure level that are regarded as relatively safe, and the duration of injury depends on the exposure time.

• Journal of Life Science
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• v.27 no.2
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• pp.187-193
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• 2017

As a part of ongoing research to elucidate and characterize antiinflammatory nutraceuticals, the crude extracts from Atriplex gmelinii C. A. Mey. and their solvent-partitioned fractions were tested for their antiinflammatory potential in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The crude extracts of A. gmelinii C. A. Mey. were fractioned according to polarity with n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and $H_2O$. Their antiinflammatory activities were investigated in LPS-induced inflammation in mouse macrophages by measuring nitric oxide (NO) generation and mRNA expression of inflammation mediators, namely, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-$1{\beta}$ ($IL-1{\beta}$), and IL-6. As a result, we confirmed that the crude extracts of A. gmelinii C. A. Mey. inhibited LPS-stimulated NO production and mRNA expression of iNOS and COX-2 as important inflammatory factors. The inhibition of NO production through the downregulation of important inflammatory factors such as iNOS, COX-2, $IL-1{\beta}$, and IL-6 was found by treatment with all solvent-partitioned fractions. Among all tested fractions, 85% aq. MeOH showed the strongest antiinflammatory response. Based on the current results, A. gmelinii C. A. Mey. was suggested to possess natural antiinflammatory components, indicating that it could be used as a valuable source of antiinflammatory substances.

• Journal of Food Hygiene and Safety
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• v.20 no.2
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• pp.118-122
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• 2005

Immu-Forte (Dong-Ahm Bio's. Corp., Korea) was evaluated fir its effectiveness as a nonspecific immunostimulator in mice. The effects of Immu-Forte were determined by analysis of cytokines using ELISh and phenotype of leukocyte subpopulations using monoclonal antibodies specific to mouse leukocyte differentiation antigens and flow cytometry. CD4 T cells, CD8 T cells, macrophages, IL-12 and IFN-r in Immu-Forte EX-treated middle dose group increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. All T cells, all B cells, macrophages, IL-2, IL-4 and IL-12 in Immu-Forte EX-treated low dose uoup increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte soy-treated group, CD4 T cells, IL-2, IL-4 and IL-12 were significantly higher in high dose-treated group, and CD 4 T cell, macrophages, IL-2, IL-4 and IL-12 were significantly higher in middle dose-treated group, and all T cell, IL-2, IL-4 and IL-12 were significantly higher in low dose-treated group. In the Itnmu-Forte A-treated group, macrophages, m cells and IL-12 in high dose-treated group and all T cells, macrophages, NK cells, IL-2, IL-4 and IL-12 in middle dose-treated group and NK cells in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte F-treated Group, all B cells, IL-4 and IL-12 in high dose-treated group and all T cells, aBl B cells, CD 4 T cells, CD8 T cells, macrophage, IL-2, IL-4, IL-12 and IFN-r in middle dose-treated group and NK cells and IL-12 in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In conclusion, the study has demonstrated that Immu-Forte had an immunostimulatory effect on mice through proliferation and activation of mouse immune cells.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.823-834
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• 1998

Background: Chronic inhalation of silica induces the lung fiborsis. The alveolar macrophages ingest the inhaled silica; they liberate the pro-inflammatory cytokines such as IL-1$\beta$, IL-6, TNF-$\alpha$ and fibrogenic cytokines, TGF-$\beta$ and PDGF. Cytokines liberated from macrophage have pivotal role in pulmonary fibrosis. There is a complex cytokine network toward fibrosis. However, the exact roles and the interaction among the proinflammatory cytokines and TGF-$\beta$, a fibrogenic cytokine, have not been defined, yet. In this study, we investigated silica induced IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ production and the effect of IL-1$\beta$, IL-6, TNF-$\alpha$ on the production of TGF-$\beta$ from lung macrophages of Balb/C mice. Method: We extracted the lung of Balb/C mice and purified monocytes by Percoll gradient method. Macrphages were stimulated by silica ($SiO_2$) in the various concentration for 2, 4, 8, 12, and 24 hours. The supernatants were used for the measurement of protein levels by bioassay, and cells for the levels of mRNA by in situ hybridization. Results: The production of IL-6 was not observed till 4 hours, and reached the peak levels at 8 hours after stimulation of silica. The production of TNF-$\alpha$ increased from 2 hours and reached the peak levels at 4 hours after stimulation of silica. The spontaneous TGF-$\beta$ production reached the peak levels at 24 hours. TNF-$\alpha$ upregulated the silica induced TGF-$\beta$ production. Silica induced TGF-$\beta$ production was blocked by pretreated anti-TNF-$\alpha$ antibody. In situ hybridization revealed the increased positive signals at 4 hours in IL-6, at 4 hours TNF-$\alpha$ and 12 hours in TGF-$\beta$. Conclusion: The results above suggest that silica induced the sequential production of IL-6, 1NF-$\alpha$ and TGF-$\beta$ from macrophages and TNF-$\alpha$ upregultaes the production of TGF-$\beta$ from silica-induced macrophages.

• Journal of Life Science
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• v.30 no.9
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• pp.804-812
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• 2020

The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

• Journal of Life Science
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• v.25 no.9
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• pp.993-999
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• 2015

The beetle Popillia flavosellata has been no reported its functional effects. In this study, we investigated the anti-inflammatory effect of P. flavosellata ethanol extract (PFE) on RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for the induction of inflammation. First, we examined the cytotoxicity of PFE in the RAW 264.7 cells at a concentration of 2,000 μg/ml or less. To evaluate the anti-inflammatory effects of PFE, we investigated the expression levels of proinflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and proinflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether PFE inhibited the translocation of nuclear factor kappa B (NF-κB) p65 into the nucleus in the LPS-induced RAW 264.7 cells. We found that the protein levels of TNF-α and IL-6 were decreased in the LPS-induced RAW 264.7 cells after the treatment with PFE in a dose-dependent manner. In addition, we confirmed that PFE inhibited the translocation of NF-κB p65 into the nucleus, as well as the protein expression levels of iNOS and COX-2. Accordingly, we propose that PFE exerts an anti-inflammatory effect through the down-regulation of NF-κB p65, TNF-α, IL-6, iNOS, and COX-2 via the toll like receptor (TLR)-4 inflammatory signaling pathway.

• The Korean Journal of Mycology
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• v.29 no.1
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• pp.15-21
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• 2001

This study was conducted to investigate the immunomodulatory activities of proteoglycan extracted from cultured mycelia of Ganoderma lucidum IY009. The proteoglycan contained two polymer peaks, one was the higher MW peak of 2,000 kD and the other was low peaks of 12kD. To understand the part of strong pharmaceutical activity between two peak, the proteoglycan was separated by ultrafiltration and column chromatography and then examined the various pharmaceutical effects. High molecular weight fraction possesing high content of ${\beta}-linked$ glucan was exhibited high antitumor activity, against sarcoma 180 bearing ICR mouse. And also, anticomplementary activity was highly observed in high molecule fraction than low it fraction. When the raw 264.7 and murine peritoneal macrophage treated with low fraction, high fraction and other stimuli. The activities inducing tumor necrosis factor of the high factions were $2.2{\sim}2.5$ times stronger than that of low fraction.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.371-379
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• 2012

This study was designed to explore new nutraceutical and cosmetic resources possessing biological activities from the plant kingdom. To fulfill this purpose, we analyzed the anti-oxidative, anti-melanogenic, and anti-inflammatory activities of Zanthoxylum schinifolium extract (ZSE) and its solvent fractions using in vitro assays and cell culture model systems. Three kinds of ZSE treated with methanol, ethanol, and water exhibited potent anti-oxidative activities through DPPH radical scavenging capacity, and inhibited in vitro DOPA oxidation. Furthermore, Z. schinifolium methanol extract (ZSME) inhibited the ${\alpha}$-melanocyte stimulating hormone, which induces melanin contents in B16F10 cells. Its anti-melanogenic activity originates from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Moreover, lipopolysaccharide induced nitric oxide production in the RAW 264.7 cell line was also ameliorated by ZSME treatment in a dose dependent manner. Among the four solvent fractions of ZSME treated with dichloromethane, ethyl acetate, n-butanol, and water, three fractions, except water, showed significant anti-melanogenic and anti-inflammatory activities. Taken together, these results provide important new insights into Z. schinifolium, indicating that it possesses numerous biological activities such as anti-oxidative, anti-melanogenic, and anti-inflammatory activities. Therefore, it may well serve as a promising material in the field of nutraceuticals and cosmetics.