• 대한한방부인과학회지
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• 제22권2호
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• pp.26-42
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• 2009

Purpose: This study was performed to evaluate the anti-inflammatory effect of Manbunbang extract (MBB). Methods: In order to understand the mechanism of anti-inflammatory effect of MBB, expression of cytokines and its levels in RAW 264.7 cell lines, as well as changes of cytokine gene expressions in serum, spleen, and liver tissues in acute inflammation induced mouse model were investigated. Results: 1. MBB significantly suppressed the expression levels of IL-1${\beta}$, TNF-${\alpha}$ and COX-2 mRNAs at 100 and 50 ${\mu}$g/m${\ell}$ concentrations, and IL-6 and NOS-II genes at 100, 50 and 10 ${\mu}$g/m${\ell}$ concentrations in RAW 264.7 cell lines, compared to those of the control. 2. MBB significantly reduced the production level of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ at 100 and 50 ${\mu}$g/m${\ell}$ concentrations in RAW 264.7 cell lines compared the those of the control. 3. MBB significantly reduced the production of IL-1${\beta}$, IL-6 and TNF-${\alpha}$ levels in sera of acute inflammation induced mice. 4. MBB significanlty suppressed the expression level of IL-1${\beta}$, TNF-${\alpha}$ mRNA in spleen tissues as well as IL-6 mRNA in liver tissues in acute inflammation induced mice. Conclusion: From the results above, anti-inflammatory effect of MBB through its immune regulation could be experimentally explained. Wide treatment of inflammatory diseases such as pelvic inflammation using MBB are recommended.

• BMB Reports
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• 제33권5호
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• pp.402-406
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• 2000

In order to understand the role of AGP on the differentiation of promyelocytic leukemia cells, the AGP expression and its relation to cytokines were investigated during granulocytic or monocytic differentiation of HL-60 cells. When HL-60 cells were treated with all-trans-retinoic acid (ATRA) for 5 days, the cells were fully differentiated into granulocytes, and the AGP mRNA and protein levels were continuously increased up to 5 days in a dose- and time- dependent manner. However, in the case of the monocytic differentiation of HL-60 cells by tetradeanoyl phorbol acetate (TPA), the AGP gene expression was not induced. In addition, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNAs were also enhanced during granulocytic differentiation. These cytokine transcripts showed a peak level 3 days after the ATRA treatment. It decreased gradually thereafter. However, direct addition of recombinant cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) and dexamethasone to the HL-60 cell cultures showed no AGP induction. These findings suggest that the AGP and proinflammatory cytokines are expressed in ATRA-treated promyelocytic cells. However, these cytokines do not act as autocrine inducers on AGP expression. This fact implies that the AGP expression during granulocytic differentiation of HL-60 cells is induced through a signal pathway different from hepatocyte signaling in inflammation.

• 동의생리병리학회지
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• 제34권6호
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• pp.319-325
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• 2020

Mahwanghangingamchosukgo-tang (MH) is recorded as a treatment to treat exterior-related respiratory diseases in the Korean medicine. In this study, we examined the anti-inflammatory effects of MH, using MH water extract and lipopolysaccharide (LPS)-induced RAW 264.7 cells. First of all, we measured the amount of nitric oxide (NO) and prostaglandin E2 (PGE2), the products of inflammatory metabolism. Also, we measured enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), as well as cytokines such as interleukin 6 (IL-6), interleukin 1 alpha (IL-1α), and interleukin 1 beta (IL-1β). MH suppressed the production of NO and PGE2 in a dose dependent manner and reduced the amount of protein and the mRNA expression of iNOS and COX-2. Also, MH reduced the mRNA expression of IL-6, IL-1α and IL-1β. In conclusion, MH decreased production of LPS-induced inflammatory factor, which could be a clinical basic subject for inflammatory diseases.

• Tuberculosis and Respiratory Diseases
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• 제73권6호
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• pp.312-319
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• 2012

Background: Muscle wasting in sepsis is associated with increased proteolysis. Interleukin-15 (IL-15) has been characterized as an anabolic factor for skeletal muscles. Our study aims to investigate the role of IL-15 in sepsis-induced muscle atrophy and proteolysis. Methods: Mice were rendered septic either by cecal ligation and puncture or by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg i.p.). Expression of IL-15 mRNA and protein was determined by reverse transcriptase polymerase chain reaction and Western blot analysis in the control and septic limb muscles. C2C12 skeletal muscle cells were stimulated in vitro with either LPS or dexamethasone in the presence and absence of IL-15 and sampled at different time intervals (24, 48, or 72 hours). IL-15 ($10{\mu}g/kg$) was intraperitoneally administered 6 hours before sepsis induction and limb muscles were sampled after 24 hours of sepsis. Cathepsin L activity was determined to measure muscle proteolysis. Atrogin-1 and muscle-specific ring finger protein 1 (MuRF1) expressions in limb muscle protein lysates was analyzed. Results: IL-15 mRNA expression was significantly lower in the limb muscles of septic mice compared to that of controls. Cathepsin L activity in C2C12 cells was significantly lower in presence of IL-15, when compared to that observed with individual treatments of LPS or dexamethasone or tumor necrosis factor ${\alpha}$. Further, the limb muscles of mice pre-treated with IL-15 prior to sepsis induction showed a lower expression of atrogin-1 and MuRF1 than those not pre-treated. Conclusion: IL-15 may play a role in protection against sepsis-induced muscle wasting; thereby, serving as a potential therapeutic target for sepsis-induced skeletal muscle wasting and proteolysis.

• 혜화의학회지
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• 제13권2호
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• pp.131-146
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• 2004

In the present study, we exarnined anti-allergic effect of SGBHB in cultured B cells. B cells were prepared from isolated murine splenocytes and activated by co-treatment of anti-CD40 monoclonal antibody and recombinant IL-4 allergens. Anti-allergic effects of SGBHB in activated B cells were determined by measuring B cell surface activated molecules (CD23+ and CD11a+), and expression levels of IL-$1{\beta}$, IL-6, IL-10, TNF-$\alpha$, IgE, and HRF. The major findings are summarized as follows. 1. SGBHB treatment did not produce significant cytotoxic effects on mouse lung fibroblast cells. 2. SGBHB produced significant inhibitory effect on the expression of B cell surface activated molecules (CD23+ and CD11a) in activated B cells. 3. SGBHB treatment significantly inhibited expression levels of IL-$1{\beta}$, IL-6, and TNF-$\alpha$ mRNAs in activated B cells.IL-6 protein levels were significantly decreased by $100{\mu}g/m{\ell}$ of SGBHB treatrrient, and TNF-$\alpha$ protein levels were decreased compared to the control group, but statistically insignificant. 4. SGBHB treatment significantly increased IL-10 at both mRNA and protein levels in activated B cells. 5. SGBHB treatment significantly inhibited levels of IgE production. Thus, the present data suggest that SGBHB has an anti-allergic effect on activated B cells by controlling irnmune responses, and further implicates the possibility on clinical application as a therapeutic agent.

• Journal of Veterinary Science
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• 제24권3호
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• pp.37.1-37.14
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• 2023

Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.

• BMB Reports
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• 제52권6호
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• pp.373-378
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• 2019

The nucleotide-binding and oligomerization domain (NOD) is an innate pattern recognition receptor that recognizes pathogen- and damage-associated molecular patterns. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) is a matrix degradation product found in the synovial fluids of patients with osteoarthritis (OA). We investigated whether NOD2 was involved in 29-kDa FN-f-induced pro-catabolic gene expression in human chondrocytes. The expression of mRNA and protein was measured using quantitative real-time polymerase chain reaction (qrt-PCR) and Western blot analysis. Small interfering RNAs were used for knockdown of NOD2 and toll-like receptor 2 (TLR-2). An immunoprecipitation assay was performed to examine protein interactions. The NOD2 levels in human OA cartilage were much higher than in normal cartilage. NOD1 and NOD2 expression, as well as pro-inflammatory cytokines, including interleukin-1beta (IL-$1{\beta}$) and tumor necrosis factor-alpha (TNF-${\alpha}$), were upregulated by 29-kDa FN-f in human chondrocytes. NOD2 silencing showed that NOD2 was involved in the 29-kDa FN-f-induced expression of TLR-2. Expressions of IL-6, IL-8, matrix metalloproteinase (MMP)-1, -3, and -13 were also suppressed by TLR-2 knockdown. Furthermore, NOD2 and TLR-2 knockdown data demonstrated that both NOD2 and TLR-2 modulated the expressions of their adaptors, receptorinteracting protein 2 (RIP2) and myeloid differentiation 88, in 29-kDa FN-f-treated chondrocytes. 29-kDa FN-f enhanced the interaction of NOD2, RIP2 and transforming growth factor beta-activated kinase 1 (TAK1), an indispensable signaling intermediate in the TLR-2 signaling pathway, and activated nuclear factor-${\kappa}B$ (NF-${\kappa}B$), subsequently leading to increased expressions of pro-inflammatory cytokines and cartilage-degrading enzymes. These results demonstrate that 29-kDa FN-f modulated pro-catabolic responses via cross-regulation of NOD2 and TLR-2 signaling pathways.

• Tuberculosis and Respiratory Diseases
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• 제72권3호
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• pp.275-283
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• 2012

Background: Healthy individuals who develop nontuberculous mycobacteria (NTM) lung disease are likely to have specific susceptibility factors which can lead to a NTM infection. The aim of the present study was to investigate the mechanism underlying innate immune responses, including the role of mitogen-activated protein kinase (MAPK), in Mycobacterium abscessus lung disease. Methods: Extracellular signal-regulated kinase (ERK1/2) and p38 MAPK expression in monocytes from peripheral blood mononuclear cells were measured by Western blot analysis after stimulation by Mycobacterium avium in five patients with M. abscessus lung disease and seven healthy controls. A M. avium-induced cytokine assay was performed after inhibition of ERK1/2 and p38 MAPK pathways. Results: Mycobacterium avium induced p38 and ERK1/2 expression in monocytes from healthy controls and subsequently upregulated tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-10 production. In monocytes from patients with M. abscessus lung disease, however, induction of p38 and ERK1/2 expression, and the production of TNF-${\alpha}$, IL-6, and IL-10 were significantly lower. Conclusion: Decreased activity of MAPK and cytokine secretion in monocytes from patients with M. abscessus lung disease may provide an explanation regarding host susceptibility to these uncommon infections.

• 한방안이비인후피부과학회지
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• 제18권3호
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• pp.1-17
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• 2005

In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis of lung disease. This Experiment was conducted to investigate the effects of Sochungyong-tang on gene expressions in Mouse Alveolar Macrophage. Fer this purpose, we observed the cytokines ($IL-1{\beta}$, IL-6, IL-10, iNOS, $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma},\;TGF-{\beta},\;TNF-{\alpha}$). We picked the alveolar macrophage out of mice and cultured it. We analyzed the cytokine gene expression by reverse transcription-PCR. The results obtained were as follows : 1 . Sochungyong-tang showed inhibitory effects on $IL-1{\beta}$ in time and concentration. 2. Sochungyong-tang showed inhibitory effects on IL-6 in time and concentration. 3. Sochungyong-tang showed inhibitory effects on IL-10 in concentration. 4. Sochungyong-tang showed inhibitory effects on iNOS. 5. Sochungyong-tang showed inhibitory effects on $TGF-{\beta}$ in time and concentration. 6. Sochungyong-tang showed on inhibitory effects on $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma}$, $TCF-{\beta}$, $TNF-{\alpha}$. According to above results, it is supposed that Sochungyong-tang has the inhibitory effects on cytokine gene expression in mouse alveolar macrophage and can be usefully applied for curing inflammatory process of lung disease. Advanced studies are required to investigate the cure mechanism of Sochungyong-tang in the future.

• 약학회지
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• 제48권2호
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• pp.159-164
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• 2004

Primary pro-inflammatory cytokines are a trio: tumor necrosis- $\alpha$ (TNF-$\alpha$), interleukine-$\beta$ (IL-$\beta$), and interleukine-6 (IL-6). These cytokines initiate and regulate the acute-phase inflammatory response during infection, trauma, or stress and appear to play an important role in the immune process. Nitric oxide (NO) is a multi-functional mediator, which plays an important role in regulating various biological functions in vivo. NO production by inducible nitric oxide synthase (iNOS) in macrophages is essential for the defense mechanisms against microorganisms and tumor cells. However, over-expression of iNOS by various stimuli, resulting in over-production of NO, contributes to the pathogenesis of septic shock and some inflammatory and auto-immune disease. Solvent fractions of sage ( Salvia officinalis L.), which is cultivated in Jeju-Do, was assayed for their effects on TNF-$\alpha$ and IL-6 production in LPS-stimulated RAW 264.7 macrophages. Hexane and ethylacetate (EtOAc) fraction of sage inhibited the protein and mRNA expression of TNF-$\alpha$ and IL-6 in LPS stimulated RAW 264.7 cells at the concentration of 100 $\mu\textrm{g}$/$m\ell$. Also, incubation of RAW 264.7 cells with the fraction of hexane or EtOAc (50 $\mu\textrm{g}$/$m\ell$) inhibited the LPS induced nitrite accumulation and the LPS/IFN-${\gamma}$ induced iNOS protein. And this inhibition of iNOS protein is concordant with the inhibition of iNOS mRNA expression. Above results suggest that extract of sage may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines (TNF-$\alpha$, IL-1$\beta$, IL-6), iNOS and NO.