• Molecules and Cells
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• v.44 no.12
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• pp.893-899
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• 2021

BLT2 is a low-affinity receptor for leukotriene B₄, a potent lipid mediator of inflammation generated from arachidonic acid via the 5-lipoxygenase pathway. The aim of this study was to investigate whether BLT2 plays any role in sepsis, a systemic inflammatory response syndrome caused by infection. A murine model of cecal ligation and puncture (CLP)-induced sepsis was used to evaluate the role of BLT2 in septic inflammation. In the present study, we observed that the levels of ligands for BLT2 (LTB₄ [leukotriene B₄] and 12(S)-HETE [12(S)-hydroxyeicosatetraenoic acid]) were significantly increased in the peritoneal lavage fluid and serum from mice with CLP-induced sepsis. We also observed that the levels of BLT2 as well as 5-lipoxygenase (5-LO) and 12-LO, which are synthesizing enzymes for LTB₄ and 12(S)-HETE, were significantly increased in lung and liver tissues in the CLP mouse model. Blockade of BLT2 markedly suppressed the production of sepsis-associated cytokines (IL-6 [interleukin-6], TNF-α [tumor necrosis factor alpha], and IL-1β [interleukin-β] as well as IL-17 [interleukin-17]) and alleviated lung inflammation in the CLP group. Taken together, our results suggest that BLT2 cascade contributes to lung inflammation in CLP-induced sepsis by mediating the production of inflammatory cytokines. These findings suggest that BLT2 may be a potential therapeutic target for sepsis patients.

• The Korea Journal of Herbology
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• v.37 no.1
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• pp.41-50
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• 2022

Objectives : The aim of this study was to investigate the effect of water extract of Agastache rugosa (AR) on productions of inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. Methods : Cell viabilities were measured with MTT assay. The production of nitric oxide (NO) from RAW 264.7 cells was measured with Griess reagent assay. The production of cytokines in RAW 264.7 cells was measured with multiplex cytokine assay. Results : AR showed no cytotoxicity on RAW 264.7 cells. AR at concentrations of 25, 50, 100, and 200 ㎍/mL significantly inhibited NO production in LPS-stimulated RAW 264.7 cells. AR at concentrations of 50, 100, and 200 ㎍/mL significantly inhibited productions of TNF-α and IL-1β in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50 and 200 ㎍/mL significantly inhibited productions of RANTES (CCL5) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 100 ㎍/mL significantly inhibited productions of macrophage inflammatory protein-1β in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50, 100, and 200 ㎍/mL significantly increased productions of IP-10 (CXCL10) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 100 and 200 ㎍/mL significantly increased MCP-1 (CCL-2) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50 and 100 ㎍/mL significantly increased productions of IL-10 in LPS-stimulated RAW 264.7 cells. Conclusions : AR might have immunomodulatory effects on productions of NO, cytokines, and chemokines in LPS-stimulated RAW 264.7 mouse macrophages.

• The Korea Journal of Herbology
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• v.35 no.2
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• pp.7-14
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• 2020

Objectives : Pyeongwi-san is widely used in Korean medicine for acute indigestion or gastrodynia. As a therapeutic agent for digestive diseases of modern people, in order to confirm the mechanism of Pyeongwi-san on digestive tract disease and the difference of therapeutic efficacy between its formulation, a comparative efficacy test was conducted on digestive tract disease animal model. Methods : For LPS enteritis animal model, male SD rats were intraorally treated with different formulation types of Pyeongwi-san, and then intraperitoneally administered LPS one hour later to induce enteritis. After 5 hours, blood was collected and TNFα, IL-1β, IL-6 and PGE2 were confirmed by ELISA. For acute gastritis animal model, male SD rats were intraorally treated with different formulation types of Pyeongwi-san according to the prescribed concentration, and then intraorally administered 60% ethanol and 150 mM HCl one hour later to induce acute gastritis. After 5 hours, blood was collected and TNFα,IL-6 were confirmed by ELISA. Results : In the LPS-administered enteritis animal model, Pyeongwi-san decreased TNFα, IL-1β, PGE2 and especially IL-6. Pyeongwi-san also decreased IL-6 in acute gastritis animal model. In addition, there was no significant difference in efficacy between the two formulations when compared with inflammatory markers. Conclusions : The efficacy of Pyeongwi-san was confirmed in the inflammatory markers related to digestive inflammatory diseases, and the efficay between two formulations of Pyeongwi-san was relatively similar. Further studies are needed to investigate the new applicability of Pyeongwi-san on different inflammatory diseases that have similar inflammation markers identified in this experiment.

• Journal of Veterinary Science
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• v.24 no.3
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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.

• Parasites, Hosts and Diseases
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• v.61 no.2
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• pp.138-146
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• 2023

Toxoplasma gondii is an intracellular protozoan parasite which can infect most warm-blooded animals and humans. Among the different mouse models, C57BL/6 mice are more susceptible to T. gondii infection compared to BALB/c mice, and this increased susceptibility has been attributed to various factors, including T-cell responses. Dendritic cells (DCs) are the most prominent type of antigen-presenting cells and regulate the host immune response, including the response of T-cells. However, differences in the DC responses of these mouse strains to T. gondii infection have yet to be characterized. In this study, we cultured bone marrow-derived DCs (BMDCs) from BALB/c and C57BL/6 mice. These cells were infected with T. gondii. The activation of the BMDCs was assessed based on the expression of cell surface markers and cytokines. In the BMDCs of both mouse strains, we detected significant increases in the expression of cell surface T-cell co-stimulatory molecules (major histocompatibility complex (MHC) II, CD40, CD80, and CD86) and cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-12p40, IL-1β, and IL-10) from 3 h post-T. gondii infection. The expression of MHC II, CD40, CD80, CD86, IFN-γ, IL-12p40, and IL-1β was significantly higher in the T. gondii-infected BMDCs obtained from the C57BL/6 mice than in those from the BALB/c mice. These findings indicate that differences in the activation status of the BMDCs in the BALB/c and C57BL/6 mice may account for their differential susceptibility to T. gondii.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.7.1-7.14
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• 2022

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

• Korean Journal of Exercise Nutrition
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• v.23 no.4
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• pp.14-22
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• 2019

[Purpose] Here, we aimed to determine the effect of Polygonum cuspidatum stem extract (PSE) on exorbital lacrimal gland-excised rat models and hyperosmotic stress-stimulated human conjunctival cells (HCCs). [Methods] Seven week old male Wistar rats were divided into six groups. Only the rats in the control group (NOR, n=5) did not undergo surgery. Three days after the surgery, the exorbital lacrimal gland-excised rats were randomly allocated to five groups: (1) vehicle-treated dry-eyed rats (DED, n=5); (2) PSE (10 mg/kg) treated DED rats (PSE-10, n=5); (3) PSE (100 mg/kg) treated DED rats (PSE-100, n=5); and (4) PSE (250 mg/kg) treated DED rats (PSE-250, n=5). In addition, the HCC line was co-treated with hyperosmolar media (528 mOsm) and PSE (1-100 μg/ml). [Results] PSE treatment restored the tear volume and goblet cell density by inhibiting severe corneal irregularities and damage. The treatment with PSE significantly attenuated the hyperosmolar stress-induced inflammation and cell death through the suppression of mRNA expression levels of Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Interleukin-1β (IL-1β), and Interferon-γ (IFN-γ), and the expression of Bcl-2-associated X protein (Bax) as well as the activation of caspase-3 in vitro. [Conclusion] The inhibitory effects of PSE treatment on dry eye disease indicate the potential of nutritional intervention by PES against inflammatory diseases without adverse effects.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1390-1401
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• 2023

In this study, a 12-week feeding experiment was conducted to characterize the effects of exogenous α-amylase on the growth, feed utilization, digestibility, plasma α-amylase activity, feed degradation rate, and fecal particle size of olive flounder (Paralichthys olivaceus). Diet was supplemented with 0 (AA₀; control), 100 (AA₁₀₀), 200 (AA₂₀₀), or 400 (AA₄₀₀) mg/kg of α-amylase, respectively. Fish (273.1 ± 2.3 g) were stocked into 12 tanks (25 fish/1,000-L tank) and 3 tanks were randomly selected for each diet group. As a result, α-amylase was found to have no significant effects (p ≥ 0.05) on the growth, feed utilization parameters, and whole-body proximate compositions. α-Amylase-treated fish exhibited only a significant increase in the apparent digestibility coefficient of carbohydrates compared to the controls. In addition, in vitro analyses revealed that α-amylase dose-dependently increased (p < 0.05) the feed degradation rate, while photographs of the intestinal content after 2, 4, and 8 h of feeding demonstrated an improved degradation rate in the α-amylase-treated groups. Plasma α-amylase content was higher in the AA₂₀₀ and AA₄₀₀ groups, whereas the control group produced significantly larger-sized fecal particles (90% size class) than these two groups. In the intestine, no changes were observed in the expression levels of the immune-related TNF-α, IL-1β, IL-2, immunoglobulin-M, HSP-70, lysozyme, and amylase alpha-2A. However, growth-related genes IGF-1, IGF-2, TGF-β3, and growth hormone genes were upregulated in muscle tissues. Collectively, exogenous α-amylase has positive roles in the modulation of the digestibility coefficient, blood α-amylase concentration, growth-related gene expression, and diet degradation for improved digestion in olive flounder.

• Journal of Microbiology and Biotechnology
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• v.34 no.4
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• pp.765-773
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• 2024

Ozone, a highly reactive oxidant molecule, is widely used as a complementary therapy for various skin diseases, including wound healing, pressure ulcers, diabetic foot, and infections. However, there is limited research on the effectiveness of ozone for atopic dermatitis (AD). Ozonated sunflower oil (OSO) is an active ingredient obtained from partially ozonated sunflower oil (SO). OSO markedly reduced the LPS-induced increase in IL-1β and nitric oxide (NO) levels in RAW 264.7 mouse macrophage cells. Oxazolone (OXZ) was applied to hairless mice to induce AD-like skin symptoms and immune response. OSO significantly alleviated the OXZ-induced increases in the number of infiltrating mast cells, epidermal thickness, AD symptoms, thymic stromal lymphopoietin (TSLP), and filaggrin, as well as the serum levels of NO, IgE, IL-1β, and TNF-α. Furthermore, OSO inhibited the IL-4/STAT3/MAPK pathway and the expression of NF-κB. Our results suggest that OSO treatment could relieve AD-mediated skin damage through its anti-inflammatory and antioxidant activities. Therefore, it can be used as a therapeutic agent against AD-related skin diseases.

• Journal of the Korean Applied Science and Technology
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• v.38 no.6
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• pp.1383-1392
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• 2021

It has been reported that emodin, a major pharmacologically active ingredient of herbal medicines such as Polygonum cuspidatum, Polygonum multiflorum, Rheum palmatum, and Aloe vera, is effective in antioxidant, antibacterial, anti-inflammatory, anticancer, and liver protection. In this study, to investigate the potential of emodin to be used as a skin disease and functional material, the activity related to the improvement of inflammation and skin barrier function was confirmed. To observe the anti-inflammatory effect on HaCaT cells, which are human keratinocytes, cytokine inhibition was confirmed by ELISA kit and protein expression by western blot. In HaCaT cells activated with TNF-α (10 ng/mL)/IFN-γ (10 ng/mL), emodin was treated with each concentration (5, 10, 20, 40) µM. As a result, It was confirmed that the production amount of TNF-α, IL-1β and IL-6 decreased as the concentration of emodin increased. In the experimental results on the expression levels of inflammation-related proteins iNOS and COX-2, it was confirmed that 48% of iNOS and 29% of COX-2 were inhibited compared to control at a concentration of 20 µM of emodin. As an indicator of skin barrier function improvement, the mRNA expression level of filaggrin, involucrin, and loricirn and the production amount of filaggrin, involucrin, and loricirn were confirmed. and excellent results were obtained with an emodin concentration-dependent increase. In particular, filaggrin, which was produced twice as much as the control at a concentration of 20 µM, is a protein involved in the formation of NMF, a natural moisturizing factor, and is known to play an important role in moisturizing the stratum corneum. In conclusion, it was confirmed that emodin can be used as a material for improving inflammation and improving skin barrier function, which is part of the potential for use as a skin disease and functional material. It is believed that if additional research is performed in the future, the scope of its application can be further expanded.