• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.291-297
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• 2015

An oral environment is constantly exposed to environmental factors and microorganisms. The periodontal ligament (PDL) fibroblasts within this environment are subject to bacterial infection and allergic reaction. However, how these condition affect PDL fibroblasts has yet to be elucidated. PDL fibroblasts were isolated from healthy donors. We examined using reverse transcription-polymerase chain reaction and measuring the intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$). This study investigated the receptors activated by exogenous bacterial pathogens (Lipopolysaccharide and peptidoglycan) and allergens (German cockroach extract and house dust mite) as well as these pathogenic mediators-induced effects on the intracellular $Ca^{2+}$ signaling in human PDL fibroblasts. Moreover, we evaluated the expression of pro-inflammatory cytokines (interleukin (IL)-$1{\beta}$, IL-6, and IL-8) and bone remodeling mediators (receptor activator of NF-${\kappa}B$ ligand and osteoprotegerin) and intracellular $Ca^{2+}$-involved effect. Bacterial pathogens and allergic mediators induced increased expression of pro-inflammatory cytokines, and these results are dependent on intracellular $Ca^{2+}$. However, bacterial pathogens and allergic mediators did not lead to increased expression of bone remodeling mediators, except lipopolysaccharide-induced effect on receptor activator of NF-${\kappa}B$ ligand expression. These experiments provide evidence that a pathogens and allergens-induced increase in $[Ca^{2+}]_i$ affects the inflammatory response in human PDL fibroblasts.

• 약학회지
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• 제51권1호
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• pp.44-50
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• 2007

Acute septic shock is one of inflammatory diseases mediated by pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. In this study, we examined the pathological difference and mechanism of lipopolysaccharides isolated from E. coli (E-LPS) or S. abortus (S-LPS) on inducing acute septic shock in ICR mouse. All mice were died by intraperitoneal treatment of S-LPS with 0.75 mg/kg, whereas E-LPS treated with even 3 mg/kg only showed 30% of mice lethal, indicating that S-LPS may be more feasible in triggering a strong septic shock condition. The secretion pattern of TNF-${\alpha}$, a critical pro-inflammatory cytokine in septic shock condition, was also distinct between E-LPS- and S-LPS-treated groups. Thus, S-LPS strikingly increased serum level of TNF-${\alpha}$ (6 ng/ml) at 1 h, while E-LPS just displayed at 2 ng/ml level. However the interaction of S-LPS with LPS receptor toll like receptor (TLR)-4, was not stronger than that of E-LPS, according to experiments with macrophage cell line RAW264.7 cells. Thus, E-LPS rather than S-LPS strongly enhanced the production of TNF-${\alpha}$. Interestingly, S-LPS more strongly up-regulated splenocyte proliferation, compared to E-LPS group, whereas there was no difference between S- or E-LPS treated groups in proliferation of Balb/c- or C57BL/6-originated splenic lymphocytes. Therefore, our data suggest that S-LPS is a more active endotoxin and that the strong septic shock-inducing effect of S-LPS seems due to the enhancement of early TNF-${\alpha}$ production and S-LPS-sensitive lymphocyte proliferation.

• 대한본초학회지
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• 제23권3호
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• pp.11-18
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• 2008

SunBangHwalMyungEum (SBH) has the effects of subduing swelling, resolving masses and alleviating pain in traditional oriental medicine. Recent studies showed that SunBangHwalMyungEum produced anti-cancer, anti-metastasis and immuno-modulatory effects. However there is lack of studies regarding the effects of SBH on the immunological activities. The present study was conducted to evaluate the effect of SBH on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Methods : After the treatment of SBH, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SBH inhibited the production of NO, iNOS, $interleukin-1{\beta}$ ($(IL-1{\beta})$), IL-6, and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with lipopolysaccharide. Conclusions : These findings suggest that SBH can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• Parasites, Hosts and Diseases
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• 제57권3호
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• pp.217-223
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• 2019

Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of $IL-1{\alpha}$, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of $IL-1{\alpha}$, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

• 대한한방부인과학회지
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• 제22권1호
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• pp.53-78
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• 2009

Purpose: In this study, we investigated the anti-thrombotic and antiinflammatory efficacy of "Gamijoukyungtang(GJKT)". Methods: We studied inhibitory effects of platelet aggregation, FXa activation, $TXB_2$ and $PGE_2$ biosynthesis and suppressive effects of GPIIb/IIIa activity and oxidative damage, pro-inflammatory cytokine reduction effects of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)' in vitro. Also, we studied suppression of pulmonary embolism, AV shunt model in rats and shortening of Rat tail bleeding time in vivo. Results: GJKT/GJKT-1 extract showed inhibitory effects on GPIIb/IIIaactivities and platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid. They suppressed biosynthesis of $PGE_2$ but GJKT-1 only supressed biosynthesis of $TXB_2$. In FXa assay, they inhibited activation of FXa. they suppressed pulmonary embolism triggered by collagen and epinephrine. In AV shunt model, they decreased the weights of AV shunt thrombus. they inhibited pro-inflammatory cytokines and decreased oxidative damages caused by DPPH. Conclusion: We confirmed the anti-thrombosis, and ant-inflammatory efficacy of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)'.

• IMMUNE NETWORK
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• 제23권3호
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• pp.25.1-25.17
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• 2023

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection induces excessive pro-inflammatory cytokine release and cell death, leading to organ damage and mortality. High-mobility group box 1 (HMGB1) is one of the damage-associated molecular patterns that can be secreted by pro-inflammatory stimuli, including viral infections, and its excessive secretion levels are related to a variety of inflammatory diseases. Here, the aim of the study was to show that SARS-CoV-2 infection induced HMGB1 secretion via active and passive release. Active HMGB1 secretion was mediated by post-translational modifications, such as acetylation, phosphorylation, and oxidation in HEK293E/ACE2-C-GFP and Calu-3 cells during SARS-CoV-2 infection. Passive release of HMGB1 has been linked to various types of cell death; however, we demonstrated for the first time that PANoptosis, which integrates other cell death pathways, including pyroptosis, apoptosis, and necroptosis, is related to passive HMGB1 release during SARS-CoV-2 infection. In addition, cytoplasmic translocation and extracellular secretion or release of HMGB1 were confirmed via immunohistochemistry and immunofluorescence in the lung tissues of humans and angiotensin-converting enzyme 2-overexpressing mice infected with SARS-CoV-2.

• Fisheries and Aquatic Sciences
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• 제21권6호
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• pp.15.1-15.9
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• 2018

Background: Inflammation has been known to associate with many human diseases. The objective of this study was to evaluate an anti-inflammatory effect of ozonated krill (Euphausia superba) oil, which was prepared by the treatment of krill oil using ozone gas. The anti-inflammatory activity was evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Results: Ozonated krill oil significantly inhibited nitric oxide (NO) production and suppressed the mRNA and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7 macrophages. Ozonated krill oil also reduced the mRNA expression of inflammatory cytokines such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor (TNF)-${\alpha}$ in LPS-stimulated RAW 264.7 macrophages. To elucidate the mechanism underlying the anti-inflammatory activity of ozonated krill oil, we evaluated the effects of ozonated krill oil on the activation of mitogen-activated protein kinases (MAPKs) pathway. Ozonated krill oil suppressed the LPS-stimulated phosphorylation of p38 MAPK and c-Jun N-terminal kinases (JNK). Conclusion: This study revealed that the ozonated krill oil exhibited an anti-inflammatory effect in LPS-stimulated RAW 264.7 macrophages. To the best of our knowledge, this is the first report that ozonated krill oil suppressed pro-inflammatory mediator and cytokine expression in LPS-stimulated RAW 264.7 macrophages by inhibiting the phosphorylation of p38 MAPK and JNK.

• Journal of Applied Biological Chemistry
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• 제62권3호
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• pp.239-246
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• 2019

In this study, an evaluation of the anti-inflammatory effect of ferulic acid isolated from Tetragonia tetragonioides in lipopolysaccharide (LPS) simulated RAW 264.7 cells was made. The chemical structure of the active compound was elucidated by $^1H$-NMR, $^{13}C$-NMR, and FAB-MS, and was confirmed to be ferulic acid. Ferulic acid was purified via open column chromatography with Sephadex LH-20 and MCI gel CHP-20. To test the anti-inflammatory effect of ferulic acid, LPS-stimulated RAW 264.7 cells were treated in subsequent experiments with different concentrations of ferulic acid (5, 10, and $25{\mu}g/mL$) and the levels of inflammatory cytokines and enzymes were also measured by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Cell viability was above 95% at acid concentrations ranging from $5-25{\mu}g/mL$. The results showed that 30% of the production of nitric oxide and 66% of prostaglandin $E_2$ were inhibited by $25{\mu}g/mL$ of ferulic acid, it also inhibited the protein expression of both inducible nitric oxide synthase and cyclooxygenase-2 by 70%. Additionally, it inhibited the production of the pro-inflammatory cytokines, tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-$1{\beta}$ by 40, 75, and 77%, respectively. According to these results, the anti-inflammatory activity of ferulic acid was demonstrated via his implication in the inhibition of the expression and secretion of inflammatory substances in LPS-stimulated RAW 264.7 cells. Therefore, we concluded that ferulic acid can be used as a functional additive having anti-inflammatory activity.

• 산업융합연구
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• 제21권8호
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• pp.69-74
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• 2023

본 논문은 Brussels Sprouts Extract의 염증성사이토카인의 억제에 미치는 영향을 살펴보기 위하여 진행되었다. 염증반응은 활성산소와 같은 매개인자와 염증성 사이토카인인 TNF-α나 IL-1β, IL-8에 의해 나타난다. 이에 본 논문은 MTS assay를 이용하여 세포에 대한 생존율을 살펴보고, RAW264.7 대식세포에 lipopolysaccharide (LPS) 자극에 의해 생성된 NO와 TNF-α나 IL-1β, IL-8과 같은 염증성 사이토카인에 대한 억제를 살펴보았다. Brussels Sprouts Extract 10mg/mL, 100mg/mL, 1000mg/mL 농도로 처리 후 억제 정도를 살펴본 결과, Brussels Sprouts Extract은 세포 독성 없이 NO생성과 TNF-α, IL-8을 농도 의존적으로 억제하였으며, 특히 1000mg/mL 농도에서는 유의한 억제를 보였다. 염증성 사이토카인의 생성을 저해한 Brussels Sprouts Extract은 염증반응 감소와 염증조절의 가능성을 시사하고 건강기능성 식품이나 염증 예방 및 치료제로서 잠재력을 제공한다고 볼 수 있다.

• 대한화장품학회지
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• 제40권1호
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• pp.45-54
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• 2014

각질형성세포주인 HaCaT세포에서 Lipopolysaccharide (LPS)로 유도하는 염증반응에는 tumor neurosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-1${\alpha}$ (IL-1${\alpha}$), interleukin-6 (IL-6), 및 interleukin-8 (IL-8)와 같은 염증 유발 사이토카인(pro-inflammatory cytokines)이 증가되고, 또한 염증반응을 매개하는 inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) 및 prostaglandins E2 (PGE2)도 증가된다. 본 연구에서는 이렇게 유도된 염증 반응에서 효과적으로 염증 관련인자를 조절할 수 있는 천연물을 발굴하고자 하였다. 그 결과, 마가목, 쐐기풀, 죽여 및 오배자 혼합 추출물이 IL-1${\alpha}$, TNF-${\alpha}$, IL-6 및 IL-8의 발현을 억제하고 COX-2, iNOS 및 PGE2의 발현도 억제함을 확인하였다. 아울러 IQ chamber$^{(R)}$을 이용하여 피부에 첩포할 때 sodium lauryl sulfate (SLS)로 유발한 피부자극에서 이들 혼합물이 효과적으로 그 자극을 억제함을 확인하였다. 따라서 본 연구 결과로부터 마가목, 쐐기풀, 죽여 및 오배자의 혼합물이 피부자극완화용 화장품 소재로서 민감성 피부 등을 위한 제품에 유용하게 사용될 수 있음을 확인하였다.