• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.2
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• pp.66-72
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• 2022

Flos Lonicerae Japonicae (the flower buds of Lonicera japonica Thunberg) has been used as an antibacterial and antiviral drug in Korean Medicine. The aim of this study is to evaluate the effect of Flos Lonicerae Japonicae water extract (FL) on the production of cytokines in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). After 24 h treatment, the production of various cytokines from RAW 264.7 was measured with multiplex cytokine assay using Bio-Plex 200 suspension array system. FL at concentrations of 50, 100, and 200 ㎍/mL significantly inhibited productions of tumor necrosis factor-α, macrophage inflammatory protein (MIP)-1β, and MIP-2 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly inhibited productions of leukemia inhibitory factor, LIX (CXCL5), and RANTES in LPS-stimulated RAW 264.7 cells; FL at concentrations of 200 ㎍/mL significantly inhibited productions of granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50 and 100 ㎍/mL significantly increased productions of interleukin (IL)-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50, 100, and 200 ㎍/mL significantly increased productions of IL-6 and interferon gamma-induced protein-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly increased productions of monocyte chemoattractant protein-1 in LPS-stimulated RAW 264.7 cells. Taken together, these data mean that FL might modulate productions of cytokines, chemokines, and growth factor in LPS-stimulated macrophages. Further study needs to verify the exact mechanism for modulatory activities of FL with macrophages.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.1
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• pp.23-29
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• 2011

Melatonin, which is the main product of the pineal gland, has well documented antioxidant and immune-modulatory effects. Macrophages produce molecules that are known to play roles in inflammatory responses. We conducted microarray analysis to evaluate the global gene expression profiles in response to treatment with melatonin in lipopolysaccharide (LPS) activated RAW 264.7 macrophage cells. In addition, eight genes were subjected to real-time reverse transcription polymerase chain reaction (RT-PCR) to confirm the results of the microarray. The cells were treated with LPS or melatonin plus LPS for 24 hr. LPS induced the up-regulation of 1073 genes and the down-regulation of 1144 genes when compared to the control group. Melatonin pretreatment of LPS-stimulated RAW 264.7 cells resulted in the down regulation of 241 genes and up regulation of 164 genes. Interestingly, among genes related to macrophage-mediated immunity, LPS increased the expression of seven genes (Adora2b, Fcgr2b, Cish, Cxcl10, Clec4n, Il1a, and Il1b) and decreased the expression of one gene (Clec4a3). These changes in expression were attenuated by melatonin. Furthermore, the results of real-time PCR were similar to those of the microarray. Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of genes involved in the regulation of immunity and defense in RAW 264.7 macrophage cells. Moreover, these results may explain beneficial effects of melatonin in the treatment of various inflammatory conditions.

• Animal cells and systems
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• v.13 no.4
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• pp.391-397
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• 2009

The house dust mite (Dermatophagoides pteronissinus) is an important factor in triggering allergic diseases. The function of eosinophils, particularly in the production of cytokine or chemokine, is critical in understanding the pathogenesis of inflammatory diseases. In this study, we examined whether D. pteronissinus extract (DpE) induces the expression of monocyte chemotactic protein 1 (MCP-1)/CCL2, IL-8/CXCL8, and IL-6 that mediate in the infiltration and activation of immune cells and in its signaling mechanism in the human eosinophilic cell line, EoL-1. DpE increased the mRNA and protein expression of MCP-1, IL-8, and IL-6 in a time- and dose-dependent course in EoL-1 cells. In our experiments using signal-specific inhibitors, we found that the increased expression of MCP-1, IL-8, and IL-6 due to DpE is associated with Src family tyrosine kinase and protein kinase C $\delta$ (PKC $\delta$). In addition, the activation of extracellular signal-regulated kinase (ERK) is required for MCP-1 and IL-8 expression while p38 mitogen-activated protein kinase (MAPK) is involved in IL-6 expression. DpE induced the phosphorylation of ERK and p38 MAPK. PP2, an inhibitor of Src family tyrosine kinase, and rottlerin, an inhibitor of PKC $\delta$, blocked the activation of ERK and p38 MAPK. DpE induces the activation of ERK and p38 MAPK via Src family tyrosine kinase and PKC $\delta$ for MCP-1, IL-8, or IL-6 production. Increased cytokine release due to the house dust mite and the characterization of its signal transduction may be valuable in understanding the eosinophil-related pathogenic mechanism of inflammatory diseases.

• Journal of Radiation Protection and Research
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• v.42 no.4
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• pp.177-188
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• 2017

Background: Radiation-induced pneumonitis and pulmonary fibrosis are common dose-limiting complications in patients receiving radiotherapy for lung, breast, and lymphoid cancers. In this study, we investigated the characteristics of effective immune cells related to pneumonitis and fibrosis after irradiation. Materials and Methods: After anesthesia, the whole thorax of C57BL/6 mice was irradiated at 14 Gy. The lung tissue and bronchoalveolar lavage fluid were collected at defined time points post-irradiation for the determination of histological and immunohistochemical analysis and inflammatory cell population infiltrated into the lung. Results and Discussion: Whole thoracic irradiation increased the deposition of extracellular matrix (ECM), lung weight, and pleural effusions, which started to die from 4 months later. At 4 months after irradiation, the numbers of macrophages and lymphocytes as well as neutrophils were increased dramatically in the lung. Interestingly, the macrophages that were recruited into the lung after irradiation had an enlarged foamy morphology. In addition, the expressions of chemokines (CCL-2, CCL-3, CXCL-10) for the attraction of macrophages and T cells were higher in the lung of irradiated mice. The high expressions of these chemokines were sustained up to 6 months following irradiation. In thoracic irradiated mice, infiltrated macrophages into the lung had the high levels of Mac-3 antigens on their surface and upregulated the hallmarks of alternatively activated macrophages such as arginase-1 and CD206. Furthermore, the levels of IL-4 and IL-13 were higher in a BAL fluid of irradiated mice. Conclusion: All results show that thoracic irradiation induces to infiltrate various inflammation-related immune cells, especially alternatively activated macrophages, through enhancing the expression of chemokines, suggesting that alternatively activated macrophages are most likely important for leading to pulmonary fibrosis.

• Molecules and Cells
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• v.43 no.11
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• pp.921-934
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• 2020

Lck-interacting transmembrane adaptor 1 (LIME) has been previously identified as a raft-associated transmembrane protein expressed predominantly in T and B lymphocytes. Although LIME is shown to transduce the immunoreceptor signaling and immunological synapse formation via its tyrosine phosphorylation by Lck, a Src-family kinase, the in vivo function of LIME has remained elusive in the previous studies. Here we report that LIME is preferentially expressed in effector T cells and mediates chemokine-mediated T cell migration. Interestingly, in LIME^-/- mice, while T cell receptor stimulation-dependent proliferation, differentiation to effector T cells, cytotoxic T lymphocyte (CTL) function and regulatory T lymphocyte (Treg) function were normal, only T cell-mediated inflammatory response was significantly defective. The reduced inflammation was accompanied by the impaired infiltration of leukocytes and T cells to the inflammatory sites of LIME^-/- mice. More specifically, the absence of LIME in effector T cells resulted in the reduced migration and defective morphological polarization in response to inflammatory chemokines such as CCL5 and CXCL10. Consistently, LIME^-/- effector T cells were found to be defective in chemokine-mediated activation of Rac1 and Rap1, and dysregulated phosphorylation of Pyk2 and Cas. Taken together, the present findings show that LIME is a critical regulator of inflammatory chemokine-mediated signaling and the subsequent migration of effector T cells to inflammatory sites.

• Biomedical Science Letters
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• v.20 no.1
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• pp.14-24
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• 2014

Hypoxia is one of the main reasons for islet apoptosis after transplantation as well as during isolation. In this study, we attempted to determine the potential usefulness of NF-${\kappa}B$ inhibitor for suppression of hypoxia-induced ${\beta}$-cell apoptosis as well as the relationship between IP-10 induction and ${\beta}$-cell apoptosis in hypoxia. To accomplish this, we cultured the mouse pancreatic ${\beta}$-cell line MIN6 in hypoxia (1% $O_2$). Among several examined chemokines, only IP-10 mRNA expression was induced under hypoxia, and this induced IP-10 expression was due to NF-${\kappa}B$ activity. Since a previous study suggested that IP-10 mediates ${\beta}$-cell apoptosis, we measured hypoxia-induced IP-10 protein and examined the effect of anti-IP-10 neutralizing Ab on hypoxia-induced ${\beta}$-cell apoptosis. However, IP-10 protein was not detected, and anti-IP-10 neutralizing Ab did not rescue hypoxia-induced MIN6 apoptosis, indicating that there is no relationship between hypoxia-induced IP-10 mRNA expression and hypoxia-induced ${\beta}$-cell apoptosis. Since it was still not clear if NF-${\kappa}B$ functions as an apoptotic or anti-apoptotic mediator in hypoxia-induced ${\beta}$-cell apoptosis, we examined possible involvement of NF-${\kappa}B$ in hypoxia-induced ${\beta}$-cell apoptosis. Treatment with 1 ${\mu}M$ NF-${\kappa}B$ inhibitor suppressed hypoxiainduced apoptosis by more than 50%, while 10 ${\mu}M$ AP-1 or 4 ${\mu}M$ NF-AT inhibitor did not, indicating involvement of NF-${\kappa}B$ in hypoxia-induced ${\beta}$-cell apoptosis. Overall, these results suggest that IP-10 is not involved in hypoxia-induced ${\beta}$-cell apoptosis, and that NF-${\kappa}B$ inhibitor can be useful for ameliorating hypoxia-induced ${\beta}$-cell apoptosis.

• IMMUNE NETWORK
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• v.13 no.6
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• pp.240-248
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• 2013

In this study, we compared the immune cell populations in rheumatoid arthritis (RA) synovial fluid, which shows lymphoid tissue-like structure, with those in tonsils, which are normal secondary lymphoid tissues. Firstly, we found that $CD4^-CD11b^+$ macrophages were the major population in RA synovial fluid and that B cells were the major population in tonsils. In addition, synovial fluid from patients with osteoarthritis, which is a degenerative joint disease, contained $CD4^+CD11b^+$monocytes as the major immune cell population. Secondly, we categorized three groups based on the proportion of macrophages found in RA synovial fluid: (1) the macrophage-high group, which contained more than 80% macrophages; (2) the macrophage-intermediate group, which contained between 40% and 80% macrophages; and (3) the macrophage-low group, which contained less than 40% macrophages. In the macrophage-low group, more lymphoid tissue inducer (LTi)-like cells were detected, and the expression of OX40L and TRANCE in these cells was higher than that in the other groups. In addition, in this group, the suppressive function of regulatory T cells was downregulated. Finally, CXCL13 expression was higher in RA synovial fluid than in tonsils, but CCL21 expression was comparable in synovial fluid from all groups and in tonsils. These data demonstrate that increased lymphocyte infiltration in RA synovial fluid is correlated with an increase in LTi-like cells and the elevation of the chemokine expression.

• Biomolecules & Therapeutics
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• v.25 no.2
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• pp.130-139
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• 2017

$CXCR5^+$ T follicular helper (Tfh) cells are associated with aberrant autoantibody production in patients with antibody-mediated autoimmune diseases including lupus. Follicular regulatory T (Tfr) cells expressing CXCR5 and Bcl6 have been recently identified as a specialized subset of $Foxp3^+$ regulatory T (Treg) cells that control germinal center reactions. In this study, we show that retroviral transduction of CXCR5 gene in $Foxp3^+$ Treg cells induced a stable expression of functional CXCR5 on their surface. The Cxcr5-transduced Treg cells maintained the expression of Treg cell signature genes and the suppressive activity. The expression of CXCR5 as well as Foxp3 in the transduced Treg cells appeared to be stable in vivo in an adoptive transfer experiment. Moreover, Cxcr5-transduced Treg cells preferentially migrated toward the CXCL13 gradient, leading to an effective suppression of antibody production from B cells stimulated with Tfh cells. Therefore, our results demonstrate that enforced expression of CXCR5 onto Treg cells efficiently induces Tfr cell-like properties, which might be a promising cellular therapeutic approach for the treatment of antibody-mediated autoimmune diseases.

• The Journal of Internal Korean Medicine
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• v.40 no.4
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• pp.567-581
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• 2019

Objective: This study aimed to evaluate the inhibitory effects of SGX01 on the lung injuries of COPD mice model. Materials and Methods: This study was carried out in two ways: in vitro and in vivo. In vitro, L929 cells were challenged with LPS, and then treated with six concentrations of SGX01 (10, 30, 50, 100, 300, and $500{\mu}g/ml$) and analyzed by ELISA. In vivo, C57BL/6 mice were challenged with LPS and cigarette smoking solution (CSS), and then treated with a vehicle only (control group), dexamethasone 3 mg/kg (dexa group), or a SGX01 200 mg/kg (SGX01 group). After sacrifice, the BALF or lung tissue was analyzed with Cytospin, FACS, ELISA, real-time PCR and H&E, and Masson's trichrome staining. Results: SGX01 significantly decreased NO, $TNF-{\alpha}$, and IL-6 on L929 cells challenged with LPS. In the COPD model, SGX01 significantly inhibited the increase of neutrophils, $TNF-{\alpha}$, IL-17A, CXCL-1, MIP2, CD8+ cells in BALF, and $TNF-{\alpha}$, $IL-1{\beta}$ mRNA expression in lung tissue. It also decreased the severity of the histological lung injury. Conclusion: This study suggests the usability of SGX01 for COPD patients by controlling lung tissue injury.

• Korean Journal of Medicinal Crop Science
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• v.27 no.3
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• pp.218-231
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• 2019

Background: We recently reported that Salvia plebeia R. Br. extracts suppress leukotriene production and effectively inhibit the airway inflammatory response by modulating inflammatory chemokine and cytokine expression. Here, we investigated the synergistic airway anti-inflammation effect of Salvia plebeia and Panax ginseng (Korean red ginseng, KRG) that has been used to treat various immune diseases such as asthma. Methods and Results: To evaluate the synergistic airway anti-inflammatory effect of Salvia plebeia and KRG, we measured the inhibitory effect of monotheraphy with either or co-theraphy with both on leukotriene and reactive oxygen species (ROS) production. Using coal a combustion, fly ash, and diesel exhaust particle (CFD)-induced respiratory disease mouse model, we found that co-theraphy synergistically suppressed airway inflammatory signs such as alveolar wall thickness and collagen fibers deposition, and decreased the number of total cell, $CD11b^+Gr-1^+$ cells, and inflammatory cytokines (IL17A, TNF, MIP-2 and CXCL-1) in bronchoalveolar lavage (BAL) fluid. Conclusions: We confirmed respiratory protection as a therapeutic effect of the Salbia plebeia-KRG 3 : 1 complex (KGC-03-PS) via anti-tracheal muscle contraction and expectorant animal studies using a CFD-induced respiratory disease mouse model.