• 대한한방부인과학회지
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• 제24권2호
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• pp.33-51
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• 2011

Objectives: This study was performed to assess the protective effect of Saengmaek-san (SM) on UVB-induced HaCaT cell damage. Methods: The protective effects of Saengmaek-san(SM) were determined by UVB-induced HaCaT assay. We assessed protective effects of Saengmaek-san (SM) on LDH release and nitrite release from HaCaT. And COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-kB, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. SM inhibited LDH-release, nitrite production in UVB-exposed HaCaT. 2. SM suppressed the gene expression of COX-2, $TNF{\alpha}$ in UVB-exposed HaCaT. 3. SM increased the gene expression of Bcl-2, Bax, Bcl-xL family protein in UVB-exposed HaCaT. 4. SM suppressed the gene expression of c-jun, c-fos, NF-kB in UVB-exposed HaCaT. Conclusions: The study showed SM inhibited the cell damage in UVB-exposed HaCaT.

• 대한치과교정학회지
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• 제33권3호
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• pp.199-210
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• 2003

치아이동 시 발생하는 골흡수에서 이미 여러 cytokine의 중요성이 강조된 바 있으며 이 가운데 interleukin-6는 구강 및 연골조직 등에서 많은 연구의 초점이 되어 왔으나 확실한 기전은 아직까지 정확히 확립되어 있지 못하다 골흡수 시 조골세포에서 유리되는 interleukin-6 (IL-6)와 nitric oxide (NO) 등이 골흡수의 조절자로 최근 대두되고 있으며 Mitogen-activated Protein kinase (MAPK)의 활성화로 인해 염증성 cytokine등이 유리될 수 있음이 최근 macrophage 등에서 증명된 바 있다. 그러므로 치아이동을 비롯한 구강 내 여러 염증의 조건에서 골흡수의 대표인자인 IL-6및 NO유리가 MAPK등의 활성 등을 통해 조절될 수 있는 가능성을 시사하고 있다. 본 연구에서 조골세포 특징을 대부분 가지고 있는 조골세포주 MC3T3El에서 p-38 MAP kinase을 매개로 NO와 IL-6가 유리됨을 확인하고자 하였다. $10\%$ Fetal Bovine Serum이 첨가된 -MEM 배양액으로 배양한 조골세포주인 MC3T3El 세포에 tumor necrosis $factor-\alpha(TNF-\alpha)$, $interferon-\gamma(IFN-\gamma)$ 및 lipopolysacchalide(LPS) 등의 단독처리 시 NO와 IL-6의 증가는 확인되지 않았으나 $TNF-\alpha/IFN-\gamma$ 혹은 $LPS/IFN-\gamma$ 등의 처치시 NO와 IL-6의 유의한 증가를 보였으며, NO발현에 직접 관여하는 inducible nitric oxide synthase (iNOS)와 IL-6 단백질 및 mRNA의 발현을 관찰하였다. 또한 specific p-38 MAP kinase inhibitor인 SB203580의 NO와 IL-6의 생성 억제를 관찰하고 단백질과 mRNA발현억제를 통해서도 확인함으로써 SB203580은 transcription 단계에서 NO와 IL-6의 생성을 조절하고 있음을 시사하여 주고 있다. $TNF-\alpha/IFN-\gamma$ 혹은 $LPS/IFN-\gamma$ 처치 시 p-38 MAP Kinase의 활성을 관찰하였으나 단독 처치 시 역시 P-38 MAP Kinase의 활성을 확인함으로써 NO와 IL-6생성기전에는 p-38 MAP Kinase이외에 다른 인자 역시 관여하고 있음을 보여주고 있다. 본 연구에서는 치아 등의 골조직의 구성 세포인 조골세포에서 NO와 IL-6유리를 확인하였으며, 또한 이들의 생성기전중의 하나로 p-38 MAP Kinase가 transcription 단계에서 관여하고 있음을 확인하였다.

• 대한본초학회지
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• 제27권6호
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• pp.23-28
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• 2012

Objectives : Samhwangsashim-tang(SHSST), a mixture of Rhei radix et rhizoma, Scutellariae radix, and Coptidis rhizoma, has been regarded as being able to treat bleeding, gastric discomfort, dry mouth, insomnia and purpura due to Blood Heat. Currently, this herbal formula is applied to gastritis, gastric ulcer, hypertension, atherosclerosis or other types of vascular inflammatory disorders. Methods : We extracted this herbal mixture with 30% ethanol and examined for its effects on systemic inflammatory responses and in vitro macrophage activity. Mice were orally given to SHSST for 7 days and then lipopolysaccharide(LPS) was intraperitoneally injected. Tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) levels in serum were measured 1 h after LPS challenge. Peritoneal macrophages were isolated from thioglycollate-injected mice and used for in vitro cellular activity. Cell death was measured using the MTT method and annexin V/propidium iodide staining. LPS-stimulated signaling molecules necessary for TNF-${\alpha}$ expression were determined by Western blotting. Results : Oral administration of SHSST for 7 days resulted in a significant reduction in LPS-stimulated TNF-${\alpha}$ release into serum. In vitro treatment of SHSST was cytotoxic in a concentration-dependent manner. However, SHSST caused a concentration-dependent reduction in necrosis and increase in apoptosis in mouse peritoneal macrophages. SHSST inhibited the activation of NF-${\kappa}B$, p38 and JNK signaling molecules in response to LPS. Conclusion : Taken together, our results demonstrated that SHSST was effective in lowering LPS-stimulated TNF-${\alpha}$ serum levels, possibly through its modulation of NF-${\kappa}B$, p38 and JNK in macrophages.

• 대한한방내과학회지
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• 제43권1호
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• pp.68-78
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• 2022

Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.

• Korean Journal of Acupuncture
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• 제28권3호
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• pp.73-83
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• 2011

Objectives : The purpose of this study is to investigate anti-allergic effects of Glycyrrhizae Radix(GR) pharmacopuncture and GR extract. Methods : In vivo, animals were gotten GR pharmacopunctures at both sides of ST36s three times for 5 days. Then, we investigated anti-DNP IgE-induced passive cutaneous anaphylaxis of Sprague Dawley rats. In vitro, we measured cell viability, ${\beta}$-hexosaminidase release, and the secretion of interleukin-4(IL-4) and tumor necrosis factor-alpha(TNF-${\alpha}$) in RBL-2H3 cells after treatment of various concentrations of GR extract. Results : In vivo, we observed inhibition of passive cutaneous anaphylaxis after GR pharmacopuncture treatments at both sides of ST36s and optional points. In vitro, GR extract treatments did not affect cell viability, but inhibited ${\beta}$-hexosaminidase release and the secretion of IL-4 and TNF-${\alpha}$. Conclusions : These results suggest that GR pharmacopuncture and GR extract should be beneficial in the inhibition of allergic inflammatory response.

• 한국임상수의학회지
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• 제25권6호
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• pp.440-446
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• 2008

Glucocorticoids (GCs) are the most widely used immunosuppressive agents, but animals treated with GCs may experience deleterious side effects which limit their use in many clinical conditions. In the present study, we examined whether methylprednisolone sodium succinate (MPSS), a glucocorticoid, modulates circulating leukocyte numbers, phagocytic capacity and oxidative burst activity (OBA) of canine peripheral blood phagocytes, and whether tumor necrosis factor-alpha (TNF-$\alpha$) release is affected by MPSS injection. Neutrophilia and monocytosis were induced by the administration of a high dose of MPSS, which is the recommended protocol for canine patients with acute spinal cord injury. The injection of MPSS decreased the phagocytic capacity of canine PMNs but not PBMCs, and recovered 12 hours (hr) after the completion of MPSS dosing. The OBA of both PMNs and PBMCs was suppressed by MPSS, and restored 24 hr after the completion of dosing. The lipopolysaccharide-induced TNF-α release by PBMCs but not PMNs exposed to MPSS was reduced 12 hr after the completion of dosing, and recovered 48 hr after the completion of dosing. These results suggest that the application of MPSS protocol inhibits the innate immune functions of canine peripheral blood phagocytes for short time relatively.

• 동의생리병리학회지
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• 제27권2호
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• pp.183-188
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• 2013

Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the inhibitory effect of MMP-1 expression of yam extract in tumor necrosis factor-alpha (TNF-${\alpha}$)-stimulated human dermal fibroblast neonatal (HDFn) cell and preventive effect of UVB-induced damage in hairless mice skin. The synthesis of procollagen and the release of MMP-1 in HDFn cells were measured by EIA kit and MMP-1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into three groups, and were topical application with the Dioscorea batatas (DB, 6%) or vehicle. Reduction of TNF-${\alpha}$-induced procollagen synthesis was increased by DB (50 ug/ml), which was higher than positive control group (TGF-${\beta}$). Also, pre-treatment of HDFn cells with DB inhibited TNF-${\alpha}$-induced release of MMP-1. In vivo study, we found that preventive effect of DB against UV-induced epidermal thickness. DB suppressed the expression of MMP-3 and MMP-13 induced by UVB irradiation. Our results show that DB have preventive effect of UV-induced skin damage in hairless mice.

• 한국식품영양과학회지
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• 제27권5호
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• pp.952-959
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• 1998

To investigate the effect of sea tangle on macrophage activity in normal and diabetic states, 10week old ICR mice were fed control(C) and sea tangle(S) diet containing 5%(w/w) cellulose and 13.6%(w/w) dry sea tangle for four weeks, after which two thirds of mice(CD and SD) were made diabetic by intramuscular injection of streptozotocin(150mg/kg bw). At 4th day after diabetes was apparent by urinary glucose, one half of diabetic mice(CDM and SDM) were treated with metformin(500mg/kg bw) orally. Peritoneal macrophages obtained from 3%-thioglycollate treated mice were cultured in the presence of lipopolysaccaride from Salmonella abortus equi(10$\mu\textrm{g}$/ml) for 24 hrs and tumor necrosis factor-$\alpha$(TNF$\alpha$), interleukin-1$\beta$(1L-1$\beta$)and prostaglandin E2(PGE2) were measured in culture media. Release of IL-1$\beta$and PGE2 from macrophage were increased in normal mice by sea tangle diet and had the same tedency in diabetic mice with or without metformin treatment although not statistically significant. Release of TNF$\alpha$ tended to be reduced by diabetes but were not changed significantly by sea tangle diet. Fatty acid compositions of macrophage and liver phospholipids showed that diabetes reduced arachidonic acid/linoleic acid ratio and sea tangle diet appeared to increase contentsof polyunsaturated fatty acids.

• 대한본초학회지
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• 제32권6호
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• pp.9-15
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• 2017

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

• 대한본초학회지
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• 제24권3호
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• pp.97-102
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• 2009

Objectives : We determined the anti-inflammatory activity of KCNS-001 that is a herbal formula including 6 medicinal plants and that are used to mitigate atopic dermatitis in oriental medicine. Methods : To evaluate anti-inflammatory effect of KCNS-001, we measured the production of reactive oxygen species (ROS), nitric oxide (NO) and cyclooxygenase-2 (COX-2) in LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay. The concentrations of ROS and relative level of NO were measured with DPPH assay and Griess reagent, respectively. COX-2 and TNF-$\alpha$ were detected by enzyme immuno assay (EIA) and enzyme-linked immunosorbent assay (ELISA). Results : ROS and NO production were reduced by KCNS-001 in a dose-dependent manner. KCNS-001 significantly inhibited activity of COX-2 and suppressed the release of tumor necrosis factor-alpha (TNF-$\alpha$). Conclusions : These results indicate that the KCNS-001 may have an anti-inflammatory agent for the treatment of various inflammatory disease.