• Journal of Microbiology and Biotechnology
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• v.27 no.10
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• pp.1827-1836
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• 2017

The world dairy industry has long been challenged by bovine mastitis, an inflammatory disease, which causes economic loss due to decreased milk production and quality. Attempts have been made to prevent or treat this disease with multiple approaches, primarily through increased abuse of antibiotics, but effective natural solutions remain elusive. Bee venom (BV) contains a variety of peptides (e.g., melittin) and shows multiple bioactivities, including prevention of inflammation. Thus, in the current study, it was hypothesized that BV can reduce inflammation in bovine mammary epithelial cells (MAC-T). To examine the hypothesis, cells were treated with LPS ($1{\mu}g/ml$) to induce an inflammatory response and the anti-inflammatory effects of BV (2.5 and $5{\mu}g/ml$) were investigated. The cellular mechanisms of BV against LPS-induced inflammation were also investigated. Results showed that BV can attenuate expression of an inflammatory protein, COX2, and pro-inflammatory cytokines such as IL-6 and TNF-${\alpha}$. Activation of NF-${\kappa}B$, an inflammatory transcription factor, was significantly downregulated by BV in cells treated with LPS, through dephosphorylation of ERK1/2. Moreover, pretreatment of cells with BV attenuated LPS-induced production of intracellular reactive oxygen species (e.g., superoxide anion). These results support our hypothesis that BV can decrease LPS-induced inflammatory responses in bovine mammary epithelial cells through inhibition of oxidative stress, NF-${\kappa}B$, ERK1/2, and COX-2 signaling.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.67-74
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• 2015

Objectives : TheHaedokgumhwa-sanwater extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages.Methods : Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE₂in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed.Results : HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells.Conclusions : Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.907-913
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• 2008

Taraxci Herba (TH; Pogongyoung in Korean) has been used in traditional oriental medicine for the treatment of various ailments. The biological activity of this plant is not yet evaluated systematically. This study was conducted to evaluate the inhibitory effects of TH on the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated Raw264.7 cells. The aim of the present work is to investigate a potential anti-inflammatory activity of TH. The Raw264.7 cells were cultured in DMEM medium for 24 h. After serum starvation for 12 h, the cells were treated with TH for 1 h, followed by stimulating NO production with LPS ($2{\mu}g/ml$). As result of this study, TH inhibited the levels of NO, PGE2, $TNF-{\alpha}$, IL-6 and $IL-1{\beta}$, and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by LPS. These inhibitory effects were mediated though the inhibition of phosphorylation of inhibitory kappa B ($I{\kappa}B$). These findings showed that TH could have some anti-inflammatory effects.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.6
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• pp.627-636
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• 2018

Endometriosis (EM) is one of the most common gynaecological disorder affecting women in their reproductive age. Mechanisms involved in the pathogenesis of EM remains poorly understood, however inflammatory responses have been reported to be significantly involved. The efficacy of 6-shogaol on proliferation of endometriotic lesions and inflammatory pathways in experimentally-induced EM model was explored in this study. EM was stimulated in Sprague-Dawley rats by implantation of autologous endometrium onto the peritoneum abdominal wall. Separate groups were treated with 6-shogaol (50, 100 or 150 mg/kg b.wt/day) via oral gavage for one month period. Gestrinone (GTN) group received GTN (0.5 mg/kg/day) as positive control. Five weeks after implantation, the spherical volume of ecto-uterine tissues was determined. Treatment with 6-shogaol significantly reduced the implant size. Histological analysis reported atrophy and regression of the lesions. 6-shogaol administration effectively down-regulated $NF-{\kappa}B$ signaling, VEGF and VEGFR-2 (Flk-1) expression in the endometriotic lesions. Excess production of $IL-1{\beta}$ and IL-6 (pro-inflammatory cytokines), PGE2 and nitric oxide (NO) were reduced. Overall, the results of the study reveal the efficacy of 6-shogaol against endometriosis via effectively suppressing proliferation of the lesions and modulating angiogenesis and $COX-2/NF-{\kappa}B$-mediated inflammatory cascades.

• Herbal Formula Science
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• v.17 no.1
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• pp.175-185
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• 2009

Petiolus Nelumbinis, branches of lotus leaf or lotus flower is a traditional oriental herbal medicine widely used for treating a superheat or disorder of qi flow. Although there are many clinical results and literature study, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of PNM (Petiolus Nelumbinis MeOH extract) in vitro. To determine cytotoxic concentration of PNM, the cells were treated with PNM for 24 h after LPS addition, and the cell viability was tested by MTT assay. Both of dosages (30 and 100 ${\mu}g/ml$) of PNM had no cytotoxicity. In these concentrations, PNM significantly reduced the elevated levels of NO and $PGE_2$ by LPS. These inhibitory effects of PNM were due to the reduced expressions of iNOS and COX-2 protein. TNF-$\alpha$, IL-1$\beta$ and IL-6 are frequently encountered pro-inflammatory cytokines, and LPS plays a key role in inducing to the massive production of these cytokines. Thus, we next determined the levels of these cytokines. Although PNM had no significant inhibitory effect on the production of TNF-$\alpha$, the elevated levels of IL-1$\beta$ and IL-6 by LPS were dose-dependently reduced in PNM-treated groups. These results demonstrate that PNM has anti-inflammatory effects by inhibiting the production of proinflammatory cytokines, NO and $PGE_2$ in LPS-activated macrophage. Moreover, the reduction of NO and $PGE_2$ levels was due to the inhibition of iNOS and COX-2 protein expression by PNM.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.2
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• pp.101-123
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• 2011

Objectives : The aim was to study the effect of bee venom pharmacopuncture therapy with different concentration on rheumatoid arthritis rat model. Methods : We enforced a bee venom pharmacopuncture therapy with different concentration on rheumatoid arthritis rat model by the intradermal injection of chicken type II collagen emulsified. 14 days after the onset of the rheumatoid arthritis rat model, a fixed volume of bee venom was daily injected to ST-35 acupoint in the rat's knee joint for 2-3 weeks. The hind paw volume, arthritic index, arthritic flexion pain test, pain threshold, and serum analysis (CRP, $PGE_2$, ALT, AST) were analyzed, and the expression profiles of COX-2, c-fos, and substance-P at the dorsal horn region of the spinal cord and subchondral bone of the knee joint were also analyzed by using the immunohistochemistry. Results : After the treatment of rheumatoid arthritis rats with bee venom pharmacopuncture, the paw volume of edema of arthritic rats were almost restored to the level of normal group, and behavior tests were very effective. Also the evaluation on the blood serum analysis was remarkable. COX-2, c-fos, and substance-P positive cells in the immunohistological section of dorsal horn region of the spinal cord and subchondral bone of the knee joints were significantly decreased. also the bee venom pharmacopuncture was effective to alleviate their rheumatoid arthritic inflammation cytokine inhibition as regards to the behavior tests and joint histological appearance. Conclusions : Based on the results in this study, bee venom pharmacopuncture with concentrated treatment condition was very effective in low fixed quantity and progressive low increased quantity.

• Biomolecules & Therapeutics
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• v.32 no.4
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• pp.467-473
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• 2024

In this study, we investigated the potential protective effects of (+)-afzelechin (AZC), a natural compound that is derived from Bergenia ligulata, on lipopolysaccharide (LPS)-induced inflammatory responses. AZC is known to have antioxidant, anticancer, antimicrobial, and cardiovascular protective properties. However, knowledge regarding the therapeutic potential of AZC against LPS-induced inflammatory responses is limited. Thus, we investigated the protective attributes of AZC against inflammatory damage caused by LPS exposure. We examined the effects of AZC on heme oxygenase (HO)-1, cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) in LPS-activated human umbilical vein endothelial cells (HUVECs). In addition, the effects of AZC on the expression of iNOS, tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were analyzed in the lung tissues of LPS-injected mice. Data revealed that AZC promoted the production of HO-1, inhibited the interaction between luciferase and nuclear factor (NF)-κB, and reduced the levels of COX-2/PGE2 and iNOS/NO, thereby leading to a decrease in the signal transducer and activator of transcription (STAT)-1 phosphorylation. Moreover, AZC facilitated the nuclear translocation of Nrf2, increased the binding activity between Nrf2 and the antioxidant response elements (AREs), and lowered the expression of IL-1β in the LPS-treated HUVECs. In the animal model, AZC significantly reduced the expression of iNOS in the lung tissue structure and the TNF-α level in the bronchoalveolar lavage fluid. These findings demonstrate that AZC possesses anti-inflammatory properties that regulate iNOS through the inhibition of both NF-κB expression and p-STAT-1. Consequently, AZC has potential as a future candidate for the development of new clinical substances for the treatment of pathological inflammation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.1
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• pp.206-213
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• 2009

Sophora flavescens, as a traditional herbal medicine, has been used to treat with a variety of disesases, In previous reports, S. flavescens and sophoraflavanone G (a prenylated flavonoid from S. flavescens) inhibited cytokines productions in LPS-induced Raw 264.7 macrophages cells and BV2 microglial cells. We examined on the anti-allergic effect of S. flavescens on the PMA plus A23187-induced rat leukemia (RBL-2H3) cells. S. flavescens inhibited the release of $\beta$-hexosaminidase and productions and expressions of tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-4 and cyclooxygenase (COX)-2 in a dose-dependent manner on stimulated RBL-2H3 cells, however, S. flavescens not affect cell viability. The protein expression level of nuclear factor (NF)-${\kappa}B$ (p65) was decreased in the nucleus and suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein, the activation of extracellular signal-regulated kinases (ERK) mitogen-activated protein kinase (MAPK) by S. flavescens. These results suggest that S. flavescens could be involved anti-allergic effect by control of $NF-{\kappa}B$ (p65) translocation into the nucleus through inhibition of $I{\kappa}B-{\alpha}$ degradation and suppression of pro-inflammatory cytokines expression.

• Journal of the Korean Applied Science and Technology
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• v.41 no.2
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• pp.251-259
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• 2024

This study evaluated the potential of hemp seed oil (Cannabis sativa L. seed oil, CSO) and hemp seed oil complex (Cannabis sativa L. seed oil complex, CSOC) as an anti-inflammatory material through comparative analysis. Anti-inflammatory effects of CSO and CSOC were confirmed through lipopolysaccharide (LPS)-induced RAW264.7 model. As a result of confirming the inhibition of lipid oxidation through lipoxygenase inhibitory activity, CSO was not inhibited, but COSC was inhibited by more than 70%. As a result of confirming cytotoxicity through MTT analysis, CSO did not show cytotoxicity, but CSOC showed cytotoxicity at over 200 ㎍/ml. In LPS-induced RAW264.7, the expression of inducible nitric oxide synthase (iNOS) and the production of nitric oxide (NO) were significantly inhibited by CSOC compared to CSO. Additionally, CSOC significantly inhibited the expression of cyclooxygenase (COX)-2 and the production of prostaglandin E₂ (PGE₂). Through this study, we confirmed that CSOC has superior anti-inflammatory effects than CSO and has the potential to be used as an anti-inflammatory material.

• Journal of Applied Biological Chemistry
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• v.61 no.3
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• pp.275-281
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• 2018

Pinus densiflora root (PDR) is used as a medicinal plant. In this study, we investigated whether the PDR extract has anti-inflammatory activities. Cell viability assays showed that the extract was not toxic toward RAW 264.7 cells at concentrations up to $10{\mu}g/mL$. At $10{\mu}g/mL$, the extract decreased nitric oxide (NO) content to 40% of the control level. The protein expression of inducible nitric oxide synthase (iNOS), which generates NO, decreased with increasing concentrations of the extract. Prostaglandin $E_2$ ($PGE_2$) levels were significantly inhibited by over 50% in the presence of $10{\mu}g/mL$ of the extract. The protein expression of cyclooxygenase-2 (COX-2), which generates $PGE_2$, decreased with increasing concentrations of the extract. Proinflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and $IL-1{\beta}$, were detected in RAW 264.7 cells after lipopolysaccharide (LPS) treatment. The extract did not affect the levels of $TNF-{\alpha}$ and IL-6, but it significantly inhibited the level of $IL-1{\beta}$. It also completely inhibited the transcription of nuclear factor-kappaB ($NF-{\kappa}B$). These results indicate that the PDR extract reduces inflammatory response-related proteins, such as NO, $PGE_2$, iNOS, and COX-2, in LPS-induced RAW 264.7 cells via the regulation of $NF-{\kappa}B$. Consequently, we have provided a mechanism to explain the anti-inflammatory effect of the PDR extract; that is, it exerts such an effect by regulating $NF-{\kappa}B$. The PDR extract can therefore be considered as an effective anti-inflammatory agent.