• Molecules and Cells
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• 제38권10호
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• pp.886-894
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• 2015

Macrophages are divided into two subpopulations: classically activated macrophages (M1) and alternatively activated macrophages (M2). BCG (Bacilli Calmette-$Gu{\acute{e}}rin$) activates disabled $na{\ddot{i}}ve$ macrophages to M1 macrophages, which act as inflammatory, microbicidal and tumoricidal cells through cell-cell contact and/or the release of soluble factors. Various transcription factors and signaling pathways are involved in the regulation of macrophage activation and polarization. We discovered that BCG-activated macrophages (BAM) expressed a new molecule, and we named it Novel Macrophage Activated Associated Protein 1 (NMAAP1). 1 The current study found that the overexpression of NMAAP1 in macrophages results in M1 polarization with increased expression levels of M1 genes, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-${\alpha}$), Interleukin 6 (IL-6), Interleukin 12 (IL-12), Monocyte chemoattractant protein-1 (MCP-1) and Interleukin-1 beta (IL-$1{\beta}$), and decreased expression of some M2 genes, such as Kruppel-like factor 4 (KLF4) and suppressor of cytokine signaling 1 (SOCS1), but not other M2 genes, including arginase-1 (Arg-1), Interleukin (IL-10), transforming growth factor beta (TGF-${\beta}$) and found in inflammatory zone 1 (Fizz1). Moreover, NMAAP1 overexpression in the RAW264.7 cell line increased cytotoxicity against MCA207 tumor cells, which depends on increased inflammatory cytokines rather than cell-cell contact. NMAAP1 also substantially enhanced the phagocytic ability of macrophages, which implies that NMAAP1 promoted macrophage adhesive and clearance activities. Our results indicate that NMAAP1 is an essential molecule that modulates macrophages phenotype and plays an important role in macrophage tumoricidal functions.

• KSBB Journal
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• 제30권1호
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• pp.38-43
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• 2015

In this study, we investigated the anti-inflammatory effects of fermented Hizikia fusiformis extracts in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The fermentation was performed using Lactobacillus casei in mixture of carbon source at $30^{\circ}C$ for 30 days. The sample groups were prepared with/without L. casei group in order to demonstrate the anti-inflammatory activity of fermented H. fusiformis in regard to lactic acid bacteria. As a result, we confirmed the inhibitory effect of H. fusiformis extracts on LPS-stimulated NO production and expression of $TNF{\alpha}$, while it had no regulatory effect on the expression of iNOS, COX-2, $IL-1{\beta}$ and IL-6 as important inflammatory factors. However, L. casei fermented group significantly suppressed the expression of the above factors. In particular, the difference between the two groups in the matter of mRNA expression of iNOS, which is directly associated with NO production, indicated that the fermentation with lactic acid bacteria effectively suppressed NO production by regulating iNOS expression. Also, effective suppression of pro-inflammatory cytokines showed that the fermentation using L. casei may provide an increment towards extraction of active ingredients that are effective anti-inflammatory agents.

• 대한한방소아과학회지
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• 제21권3호
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• pp.33-55
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• 2007

Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

• 대한한방소아과학회지
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• 제22권1호
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• pp.69-93
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• 2008

Objectives The purpose of this study is to investigate the effect of KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to this condition in humans. Methods To investigate the effect of KKHS on atopic dermatitis (AD), we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells(PBMCs), splenocytes, draining lymph node(DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis of NC/Nga mouse in vivo. Results In vivo, clinical skin severity score was significantly lower in the KKHS group than in the control group. IgE, IL-6, TNF-${\alpha}$, IgM, IgG2a and IgG2b levels in serum decreased remarkably in the KKHS group than in the control group, and the level of IFN-${\gamma}$ production which is secreted from Th1 cell was increased by KKHS. After this experiment we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+CD69^+$, $CD4^+CD25^+$ and $CD49b^+$) by flow cytometry. It results that the total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as much as normal state, and the level of $CD3^+CD69^+$ in isolated DLN and PBMCs were significantly decreased, and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by KKHS. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E), toluidine staining (mast cells marker). KKHS were very effective to the histological symptoms which are in dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration. Ear thickness was significantly decreased compared with the control group and the size of inflammatory lymphocytes cells (ILC) and plasma cells (PC) in DLN were also decreased. Conclusions KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse was very effectiveness to the atopy dermatitis treatment.

• 한국식품영양학회지
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• 제34권5호
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• pp.487-497
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• 2021

To investigate the industrial availability of liquid fermentation (PL-ferment) by Phellinus linteus mycelium as a postbiotics for the inhibition of inflammation, PL-ferment was fractionated into culture supernatant (CS), hot-water extract (HW) from PL-ferment, EtOH-precipitate (CP) fractionated from HW, and the dialysate (DCP) of CP. Compared to the other fractions, DCP which is expected to contain exopolysaccharide (EPS) as the major component, significantly decreased the production of NO, IL-6, and MCP-1 in LPS-induced RAW 264.7 cells, and IL-6 and IL-8 in TNF-α and IFN-γ-induced HaCaT cells. The general component analysis results showed that no significant difference in components was observed between the fractions, whereas sugar composition analysis revealed that DCP had decreased glucose and increased mannose contents compared to the other fractions. This suggests that mannose played an important role in the anti-inflammatory activity of the active fraction, DCP. Molecular weight distribution analysis revealed that DCP was mainly composed of low-molecular-weight material-removed high-molecular-weight polysaccharides of 18-638 kDa, suggesting that EPS originated from P. linteus EPS. In conclusion, our results suggest that the DCP of P. linteus mycelium fermentation using the anti-inflammatory activity could be used industrially as postbiotic material.

• 한국식품영양학회지
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• 제32권1호
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• pp.1-10
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• 2019

In order to examine the functionality of broccoli sprout (Brassica oleracea, BS), solvent extracts were prepared and their anti-oxidative and immunomodulating activities were compared with those of broccoli (B). EtOH extracts (E) were potently higher than hot-water extracts (HW) in the antioxidant contents and radical scavenging activity. In particular, the total polyphenolic contents in addition to ABTS and DPPH radical scavenging activity were significantly higher in EtOH extract of broccoli sprout (BS-E; 9.15 mg GAE/g, 4.52 mg AEAC/g, and 1.14 mg AEAC/g) compared with that of broccoli (B-E; 7.83 mg GAE/g, 3.63 mg AEAC/g, and 0.97 mg/AEAC/g). Whereas, total flavonoid content was significantly higher in B-E (1.60 mg QE/g) than BS-E (1.43 mg QE/g). Anti-inflammatory activity was investigated using LPS-induced cell line model at a concentration of $10{\sim}100{\mu}g/mL$, in which all solvent extracts of both broccoli sprouts and broccoli were not toxic to RAW 264.7 cell lines. In anti-inflammatory activity of broccoli sprouts, EtOH extracts also showed significantly more potent activity than hot-water extracts in all sample concentrations tested. In addition, BS-E ($100{\mu}g/mL$) inhibited nitric oxide (NO) and IL-6 production to 60.9% and 68.9% compared with the LPS inflammation group (without extracts), whereas B-E inhibited 49.6% and 54.9%. On the other hand, in immunostimulating activity by splenocytes and macrophages, hot-water extract showed significantly higher activity than EtOH extract. Especially, BS-HW stimulated the splenocyte proliferation (1.2-fold against saline group) and IFN-${\gamma}$ production (264.39 pg/mL) at $100{\mu}g/mL$, and the production of IL-6 (1.33-fold), IL-12 (1.09-fold) and TNF-${\alpha}$ (1.49-fold) from macrophages was also significantly enhanced over broccoli. In conclusion, broccoli sprouts showed more potent anti-oxidative, anti-inflammatory and immunostimulating activity than broccoli, suggesting the possibility of using broccoli sprouts as functional food materials.

• 대한한방내과학회지
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• 제43권4호
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• pp.514-528
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• 2022

Objective: To examine the effects of Trichosanthes kirilowii Maximowicz extract (TE) and Trichosanthes kirilowii Maxi mowicz Cheonghyeol Plus Phellinus linteus Cheonghyeol plus (TCP) on anti-inflammatory factor expression in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were activated with TNF-α and then treated with TE and TCP. The expression levels were then measured for intracellular genes (KLF2, eNOS, MCP-1, ICAM-1, and VCAM-1), proteins (KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, ERK, and JNK, p38), and extracellular biomarkers (ICAM-1, VCAM-1, and MCP-1). Results: 1. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2 and eNOS intracellular genes and significantly decreased the expression of ICAM-1, VCAM-1, and MCP-1 genes compared to the control group. 2. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2, eNOS proteins compared to the control group, and significantly reduced the expression of VCAM-1, ICAM-1, MCP-1, ERK, and p38 proteins. However, JNK protein phosphorylation showed no significant change compared to the control group. 3. TCP at concentrations of 100 ㎍/mL or more significantly decreased the production of MCP-1, ICAM-1, and VCAM-1 extracellular biomarkers compared to the control group. 4. TE at a concentration of 100 ㎍/mL did not cause any significant change in the expression of intracellular genes or proteins, in the production of the extracellular biomarker MCP-1, or in the amount of JNK protein compared to the control group. Other intracellular genes, proteins, and extracellular biomarker expression showed the same trend as observed with TCP exposure. Conclusion: This study experimentally confirmed that TE and TCP could be effective in preventing or inhibiting various inflammatory vascular diseases due to their anti-inflammatory effects.

• 대한본초학회지
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• 제29권2호
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• pp.39-45
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• 2014

Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

• 대한본초학회지
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• 제32권2호
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• pp.25-32
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• 2017

Objectives : This study was conducted to investigate candidate materials as anti-inflammatory agent from extracts of Korean medicinal plants in Hwaak mountain. Ligustrum obtusifolium (LO) is a Korea medicinal plants that commonly used for robustness and hemostasis. It has been reported that LO has exhibited anti-ischemic, anti-oxidative, anti-hypolipidemic, anti-tumor and hypoglycemic effects. However, LO has not been previously reported to have an anti-inflammatory effect. Therefore, we have evaluated the anti-inflammatory effects of LO and its underlying molecular mechanisms in LPS-induced RAW 264.7 macrophages. Methods : Cell viability was determined by MTT assay in RAW 264.7 macrophages. Nitric Oxide (NO) was measured with Griess reagent and pro-inflammatory cytokines were detected by ELISA in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and p65 subunit of nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) were determined by Western blot analysis. Results : Among 15 extracts of Korean medicinal plants tested, Ligustrum obtusifolium (LO) showed the inhibition of NO production without cytotoxicity. LO reduced the expression levels of iNOS and COX-2 proteins in LPS-simulated RAW 264.7 macrophages in dose-dependent manner. Consistent with these data, LO inhibited the productions of $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$ in LPS-simulated RAW 264.7 macrophages. Furthermore, LO attenuated the LPS-induced nuclear translocation of p65 $NF-{\kappa}B$ in RAW 264.7 macrophages involving suppression of $NF-{\kappa}B$ activation. Conclusions : Taken together, these results suggest that the anti-inflammatory effects of LO is associated with regulation of inflammatory mediators via inhibition of $NF-{\kappa}B$ activation in LPS-treated RAW 264.7 macrophages.

• 대한본초학회지
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• 제33권5호
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• pp.81-88
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• 2018

Objectives : The aim of this study was to investigate the protective effects of an aqueous extract from Taxillus chinensis (DC.) Danser (TCE) in Monosodium iodoacetate (MIA)-induced osteoarthritis (OA) rat model. Methods : Sprague Dawley male rats were divided into the following four groups (n=6 per group): Normal (saline control), MIA (MIA-induced OA with vehicle), TCE (MIA-induced with TCE treatment), and IM (MIA-induced with indomethacin treatment). Rats in which OA was induced by MIA were treated with TCE (200 mg/kg) or indomethacin (1 mg/kg) for 4 weeks. Weight-bearing on the hind legs and body weights were measured weekly. At the end of the experiment (3 weeks after MIA injection), serum aspartate aminotransferase and alanine aminotransferase levels were measured to assess the liver toxicity induced by TCE. Its effects on serum inflammatory cytokine levels and tissue histopathology were also evaluated. Results : TCE restored the hind limb weight-bearing distribution. Serum levels of Interleukin 6 (IL-6), Tumor necrosis factor alpha (TNF-${\alpha}$) and Leukotriene B4 (LTB4) were significantly higher in the MIA group than in the Normal group, but serum IL-6 levels were significantly lower in the TCE group. In the TCE group, the synovial membrane was protected in hematoxylin and eosin and Safranin-O staining, respectively. Conclusions : TCE recovered the hind paw weight bearing distribution, inhibited the production of inflammatory cytokine, and protected synovial tissue and cartilage in the OA rat model. Therefore, TCE appears to be an effective therapeutic agent for treating OA and OA-related symptoms.