• Journal of Sasang Constitutional Medicine
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• v.22 no.3
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• pp.141-151
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• 2010

1. Objectives: Three herbal formulas (Yuldahanso-tang, Chungsimyonja-tang, and Taeeumjowi-tang) for Taeeumin were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. 2. Methods: This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results: Yuldahanso-tang and Chungsimyonja-tang increased NO synthesis in RAW 264.7 cells. Three herbal formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<.001). Three herbal formulas for Taeeumin also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. 4. Conclusion: These results suggested that three herbal formulas for Taeeumin could be used as stimulator of immune response.

• Korean Journal of Pharmacognosy
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• v.48 no.1
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• pp.18-24
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• 2017

This study was conducted to determine the anti-inflammatory effect of essential oil extracted from the wood of Chamaecyparis obtusa Sieb. et Zucc. Endl. (Cupressaceae). The essential oil was extracted from the wood of C. obtusa by hydrodistillation method, and conducted the analysis on the chemical composition of the extracted C. obtusa wood oil through GC-MS. The major constituents of the oil were found to be: ${\alpha}-pinene$ (11.4%), cadinene (5.4%), ${\delta}-cadiene$ (9.0%), ${\tau}-muurolol$ (22.2%), ${\alpha}-cadinol$ (20.8%) etc. We attempted to identify the anti-inflammatory activities of the oil when it is injected in the lipopolysaccharide (LPS)-stimulated RBL-2H3 cells, along with its effects on the secretion of interleukin-4 (IL-4), interleukin-13 (IL-13), ${\beta}-hexosaminidase$. According to the cell viability analysis conducted by MTT assay, the oil in $10^{-7}{\sim}10^{-5}%$concentration showed no effect on the cell viability. After RBL-2H3 cells treated by LPS stimulation were exposed to $10^{-7}%$ concentration of C. obtusa wood oil, the expression levels of IL-4, IL-13 within the cell were observed to remarkably decrease. Also, it was attenuated the release of ${\beta}-hexosaminidase$ from mast cells to a significantly meaningful level. These results suggest that C. obtusa wood oil exerts the anti-inflammatory effect, by regulating the expression of inflammatory cytokines, which is a valuable feature to be highly utilized as the functional materials in the future.

• Nutrition Research and Practice
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• v.7 no.5
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• pp.352-358
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• 2013

Korean pine nut oil (PNO) has been reported to have favorable effects on lipid metabolism and appetite control. We investigated whether PNO consumption could influence weight gain, and whether the PNO-induced effect would result in an improvement of immune function in high-fat diet (HFD)-induced obese mice. C57BL/6 mice were fed control diets with 10% energy fat from either PNO or soybean oil (SBO), or HFDs with 45% energy fat from 10% PNO or SBO and 35% lard, 20% PNO or SBO and 25% lard, or 30% PNO or SBO and 15% lard for 12 weeks. The proliferative responses of splenocytes upon stimulation with concanavalin A (Con A) or lipopolysaccharide (LPS), Con A-stimulated production of interleukin (IL)-2 and interferon (IFN)-${\gamma}$, and LPS-stimulated production of IL-6, IL-$1{\beta}$, and prostaglandin $E_2$ ($PGE_2$) by splenocytes were determined. Consumption of HFDs containing PNO resulted in significantly less weight gain (17% less, P < 0.001), and lower weight gain was mainly due to less white adipose tissue (18% less, P = 0.001). The reduction in weight gain did not result in the overall enhancement in splenocyte proliferation. Overall, PNO consumption resulted in a higher production of IL-$1{\beta}$ (P = 0.04). Replacement of SBO with PNO had no effect on the production of IL-2, IFN-${\gamma}$, IL-6, or $PGE_2$ in mice fed with either the control diets or HFDs. In conclusion, consumption of PNO reduced weight gain in mice fed with HFD, but this effect did not result in the overall improvement in immune responses.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.923-929
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• 2004

Stylopine is a major component of the leaf of Chelidonium majus L. (Papaveraceae), which has been used for the removal of warts, papillomas and condylomas, as well as the treatment of liver disease, in oriental countries. Stylopine per se had no cytotoxic effect in unstimulated RAW 264.7 cells, but concentration-dependently reduced nitric oxide (NO), prostaglandin E$_2$ (PGE$_2$), tumor necrosis factor-a (TNF-$\alpha$) and interleukin-1$\beta$(IL-1$\beta$), and the IL-6 production and cyclooxygenase-2 (COX-2) activity caused by the LPS stimulation. The levels of inducible nitric oxide synthase (iNOS) and COX-2 protein expressions were markedly suppressed by stylopine in a concentration dependent manner. These results suggest that stylopine suppress the NO and PGE$_2$ production in macrophages by inhibiting the iNOS and COX-2 expressions. These biological activities of stylopine may contribute to the anti-inflammatory activity of Cheli-donium majus.

• BMB Reports
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• v.47 no.11
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• pp.649-654
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• 2014

Neutrophils play an important role in the initiation of innate immunity against infection and injury. Although many different types of G-protein coupled receptors are functionally expressed in neutrophils, no reports have demonstrated functional expression of umami taste receptor in these cells. We observed that mouse neutrophils express the umami taste receptor T1R1/T1R3 through RNA sequencing and quantitative RT-PCR analysis. Stimulation of mouse neutrophils with L-alanine or L-serine, which are ligands for the umami taste receptor, elicited not only ERK or p38 MAPK phosphorylation but also chemotactic migration. Moreover, addition of L-alanine or L-serine markedly reduced the production of several cytokines including $TNF-{\alpha}$ induced by lipopoly-saccharide (LPS) through inhibition of $NF-{\kappa}B$ activity or STAT3 phosphorylation in neutrophils. Our findings demonstrate that neutrophils express the umami taste receptor, through which tastants stimulate neutrophils, resulting in chemotactic migration, and attenuation of LPS-induced inflammatory response.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.156-161
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• 2000

The unripe fruit of Poncirus trifoliata Raf has been used for the treatment of allergic disease. Recently it was reported that the fruit inhibits passive cutaneous anaphylaxis and histamine release at mast cell. Type I immediate hypersensitivity of anaphylactic type is caused by released mediate chemical at mast cell. Histamine is also known as one of potent mediate chemical. Also release of mediate chemical is affected by specific stimulation of IgE combined with mast cell. Activation of mast cell is known to be stimulated by compound 48/80 and inhibited by increase of cAMP. In this experiment, the effect of water extract of Poncirus trifoliata Raf fruit (PT) on a histamine release, cAMP concentration and IgE production was measured. Compound 48/80 was administrated to the mouse peritoneal cell which was pretreated with PT. PT dosedependently inhibited histamine release at peritoneal mast cell and the serum level of histamine induced by compound 48/80. PT also instantly increased cAMP level of peritoneal mast cell right after it was added and the level gradually decreased. Production of IgE induced by antigens at mouse peritoneal cell was inhibited by PT. The IgE synthesis is induced by IL-4 and it is known that lipopolysaccharide(LPS) plus IL-4 cause an increase in IgE secretion by murine B cells. The effects of PT inhibited the production of IgE activated by LPS plus IL-4 at human U266B1 cells. These results indicate that PT has antiallergic activity by Inhibition of IgE production from B cells and histamine release by increase of cAMP.

• Molecular & Cellular Toxicology
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• v.5 no.3
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• pp.187-192
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• 2009

Toll-like receptors (TLRs) play an important role in host defense by sensing invading microbial pathogens. The stimulation of TLRs by microbial components triggers the activation of the myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter inducing interferon-$\beta$ (TRIF)-dependent downstream signaling pathways. TLR/MyD88 signaling pathway induces the activation of nuclear factor-kappa B (NF-${\kappa}B$) and the expression of inflammatory cytokine genes, including tumor necrosis factor-alpha, interleukin (IL)-6, IL-12, and IL-$1{\beta}$. On the other hand, TLR/TRIF signaling pathway induces the delayed-activation of NF-${\kappa}B$ and interferon regulatory factor 3 (IRF3), and the expression of type I interferons (IFNs) and IFN-inducible genes. The divalent heavy metal cadmium (Cd) is clearly toxic to most mammalian organ systems, especially the immune system. Yet, the underlying toxic mechanism(s) remain unclear. Cd inhibits the MyD88-dependent pathway by ceasing the activity of inhibitor-${\kappa}B$ kinase. However, it is not known whether Cd inhibits the TRIF-dependent pathway. Presently, Cd inhibited NF-${\kappa}B$ and IRF3 activation induced by lipopolysaccharide (LPS) and polyinosinic-polycytidylic acid. Cd inhibited LPS-induced IRF3 phosphorylation and IFN-inducible genes such as interferon inducible protein-10 and regulated on activation normal T-cell expressed and secreted (RANTES). These results suggest that Cd can modulate TRIF-dependent signaling pathways of TLRs.

• The Journal of Korean Medicine
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• v.31 no.5
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• pp.112-123
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• 2010

Objectives: Three yin-tonifying formulae (Ssanghwa-tang, Yukmijihwang-tang and Jaeumganghwa-tang) were applied to investigate their immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in the murine macrophage cell line (RAW 264.7) and in ovalbumin (OVA)-immunized mice. Methods: This study was carried out in nitricoxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes in association with three herbal formulas. C57BL/6 mice were immunized intraperitoneally with OVA/aluminum ($100\;{\mu}g/200\;{\mu}g$/mouse) on days 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. Results: All three yin-tonifying formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Also, these herbal formulas all significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. Conclusion: These results suggested that the three yin-tonifying formulae could be used as stimulators of immune response.

• The Korean Journal of Physiology
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• v.29 no.1
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• pp.1-11
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• 1995

Alveolar macrophages play a pivotal role in the pathogenesis of silicosis since the macrophages may release a wide variety of toxic and inflammatory mediators as well as mitogenic growth factors. In the present study, the effects of in vitro exposure to silica on release of various mediator such as reactive oxygen species, platelet activating factor(PAF), and interleukin-1 (IL-1) by alveolar macrophages were examined. First, hydrogen peroxide release from alveolar macrophages was monitored by measuring the change in fluorescence of scopoletin in the absence or presence of graded concentration of silica. Significantly enhanced release of hydrogen peroxide was observed at 0.5 mg/ml and above. A maximal enhancement of 10 fold above control was observed at 5 mg/ml silica. Similarly, in vitro exposure to silica also significantly stimulated the generation of chemiluminescence from alveolar macrophages at 0.5 mg/ml and above with n maximal enhancement of 8 fold at 5 mg/ml silica. Second, PAF release from alveolar macrophages after 30 min incubation at $37^{\circ}C$ in absence or presence of zymosan and silica was determined by measuring $^{3}H-serotonin$ release ability of the conditioned macrophage supernates from platelets. 5 mg/ml zymosan as a positive control fur the PAF assay increased PAF release by 19 % of total serotonin release. Furthermore, silica also resulted in significant enhancement of the PAF release compared with that in unstimulated (control) cells, i.e., $17.7{\pm}5.8%$ and $24.0{\pm}4.9%$ of total serotonin release at 5 mg/ml and 10 mg/ml silica, respectively, which represents the release of nanomole levels of PAF. Lastly, IL-1 production by alveolar macrophages was analysed following their stimulation with lipopolysaccharide (LPS) and silica by their capacity to stimulate thymocyte proliferation. $10\;{\mu}g/ml$ LPS resulted in an 11 fold increase in IL-1 production. In comparison, $50\;{\mu}g/ml$ silica resulted in a 4 fold increase in IL-1 release. These data indicate that in vitro exposure of alveolar macrophages to silica activates the release of various bioactive mediators such as reactive oxygen species, PAF and IL-1 which thus contribute to amplification of inflammatory reactions and regulation of fibrotic responses by the lung after inhalation of silica.

• Journal of Veterinary Science
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• v.24 no.1
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• pp.11.1-11.14
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• 2023

Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ⁺CD3⁺ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ⁺ CD4^-CD8⁺ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.