• International Journal of Oral Biology
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• 제43권3호
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• pp.141-146
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• 2018

Periodontitis is generally a chronic disorder characterized by breakdown of tooth-supporting tissues, producing dentition loss. Porphyromonas gingivalis (P. gingivalis), a Gramnegative anaerobic rod, is one of the major pathogens associated with periodontitis. Neutrophils are first line defense cells in the oral cavity that play a significant role in inflammatory response. Xylitol is a known anti-caries agent and has anti-inflammatory effects. In this study, we conducted experiments to evaluate anti-inflammatory effects of xylitol on P. gingivalis infected neutrophils for possible usage in prevention and treatment of periodontal infections. P. gingivalis was intraperitoneally injected and peritoneal lavage was collected for cytokine determination. For in vitro study, neutrophils were collected from mouse peritoneal cells after zymosan injection or bone marrow cells. Neutrophils were stimulated with live P. gingivalis and ELISA was used to determine the effect of xylitol on P. gingivalis induced cytokine production. $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ concentration and neutrophil population in the peritoneal lavage was increased in P. gingivalis-infected mouse. Peritoneal cells infected with live P. gingivalis revealed significantly increased production of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ at multiplicity of infection of 10. Neutrophils from bone marrow and peritoneal lavage revealed increased production of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$. Xylitol significantly mitigated P. gingivalis induced cytokine production in neutrophils. Findings indicate that xylitol is an anti-inflammatory agent in neutrophils infected with live P. gingivalis, that suggests its use in periodontitis management.

• 한방안이비인후피부과학회지
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• 제25권3호
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• pp.1-12
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• 2012

Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.438-445
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• 2010

The immunomodulatory effects of Aureohasidium pullulans SM-2001 exopolymers containing $\beta$-1,3/1,6-glucan were evaluated in cyclophosphamide (CPA)-treated mice. To induce immunosuppression, 150 and 110 mg/kg of CPA were intraperitoneally injected 3 days and 1 day, respectively, before beginning administration of the test material. Exopolymers were delivered subcutaneously or orally, four times, in a volume of 10 ml/kg at 12-h intervals beginning 24 h after the second CPA treatment. Changes in thymus and spleen weights, splenic amounts of tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1{\beta}$, and IL-10, and numbers of CD3+, CD4+, CD8+, and TNF-$\alpha+$ thymus and spleen cells were monitored in CPA-treated mice. As a result of CPA treatment, dramatic decreases in the number of CD3+, CD4+, CD8+, and TNF-$\alpha+$ cells were detected in the thymus and spleen, along with decreases in thymus and spleen weights. In addition, splenic TNF-$\alpha$, IL-$1{\beta}$, and IL-10 contents were also decreased on observation with flow cytometry. However, oral and subcutaneous treatments with exopolymers effectively reduced the immunosuppressive changes induced by CPA. Therefore, it is concluded that exopolymers of A. pullulans SM-2001 can effectively prevent immunosuppression through, at least partially, the recruitment of T cells and TNF-$\alpha+$ cells or enhancement of their activity, and can provide an effective component of prevention or treatment regimens for immunosuppression related to cancer, sepsis, and high-dose chemotherapy or radiotherapy.

• The Korean Journal of Physiology and Pharmacology
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• 제21권1호
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• pp.117-124
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• 2017

The present study aimed to show that pro-inflammatory cytokines [tumor necrosis factor (TNF)-${\alpha}$, interferon (IFN)-${\gamma}$, and interleukin (IL)-$1{\beta}$] synergistically induce the production of nitric oxide (NO) production in mouse mesangial cells, which play an important role in inflammatory glomerular injury. We also found that co-treatment with cytokines at low doses (TNF-${\alpha}$; 5 ng/ml, IFN-${\gamma}$; 5 ng/ml, and IL-$1{\beta}$; 1.25 U/ml) synergistically induced NO production, whereas treatment with each cytokine alone did not increase NO production at doses up to 100 ng/ml or 50 U/ml. Silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), attenuates cytokine mixture (TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$)-induced NO production. Western blot and RT-PCR analyses showed that silymarin inhibits inducible nitric oxide synthase (iNOS) expression in a dose-dependent manner. Silymarin also inhibited extracellular signal-regulated protein kinase-1 and -2 (ERK1/2) phosphorylation. Collectively, we have demonstrated that silymarin inhibits NO production in mouse mesangial cells, and may act as a useful anti-inflammatory agent.

• 한방안이비인후피부과학회지
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• 제22권3호
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• pp.63-70
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• 2009

Objectives : The present study was examined to evaluate the anti-inflammatory effects of the Gentianae Macrophyllae Radix MeOH extracts (GMR) in vivo. Methods : The effects of GMR on anti-inflammation were measured by production of NO, TNF-$\alpha$ (Tumor Necrosis Factor-alpha) and IL-$1{\beta}$ (Interleukin-$1{\beta}$), IL-6 in Raw 264.7 macrophage cells stimulated with LPS. Results : 1. All concentrations of GMR(0.10 mg/ml) had no significant cytotoxicity in Raw 264.7 cell during the entire experimental period. 2. The level of NO and iNOS in culture medium was dramatically increased by LPS application. However, these increases were dose-dependently(0.03 and 0.10 mg/ml) attenuated by treatment with GMR. 3. All concentrations of GMR significantly inhibited the production of IL-$1{\beta}$ in Raw 264.7 macrophage cells stimulated with LPS. Conclusions : These results provide evidences that therapeutic effect of GMR on heat syndrome, especially due to the acute inflammation, are partly due to the reduction of some of inflammatory factors by inhibiting iNOS and COX-2 through the suppression of $p-I{\kappa}B{\alpha}$. Moreover, it suggests that the mechanism of action of GMR comes from the suppression of inflammatory mediators, such as NO, PGE2 and pro-inflammatory cytokines.

• 생명과학회지
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• 제21권8호
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• pp.1176-1182
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• 2011

본 연구는 고지방식이로 유도된 비만쥐를 대상으로 저지방식이와 중강도 운동이 비장세포의 전염증성 사이토카인 발현에 미치는 영향을 보고자 하였다. 4 주령의 C57BL/6 마우스를 대상으로 고지방식이(45% fat)를 5 주간 처치하여 비만을 유도한 후, 저지방식이(10% fat)로 전환한 후 통제군(운동미실시, n=10)과 운동군(중강도운동실시, n=10)으로 나누어 8 주간 트레드밀을 이용하여 중강도 운동을 주 5회, 1 일 30-60분 실시하였다. 운동종료후 비장세포를 분리하여 Concanavalin A (10 ug/ml)로 24시간 자극 후 전염증성 사이토카인인 IL-1${\beta}$, IL-6, TNF-${\alpha}$를 Bio-Plex를 활용하여 측정하였다. 자료분석은 독립 t 검증을 실시 하였으며, 유의수준은 p<0.05 수준으로 하였다. 비장세포에서 발현된 전염증성 사이토카인인 IL-1${\beta}$, IL-6, TNF-${\alpha}$는 저지방식이를 단독처치한 통제군에 비하여 중강도 운동을 병행실시한 운동군에서 유의하게 감소한 결과(p<0.01)를 나타내어 고지방식이로 유도된 비만쥐에서 저지방식이의 단독처치보다 중강도 운동의 병행 처치가 전염증 사이토카인 감소에 긍정적인 영향을 미치는 것으로 사료된다.

• 한국식품영양학회지
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• 제29권3호
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• pp.431-437
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• 2016

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-${\kappa}B$의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다.

• 한국한의학연구원논문집
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• 제16권3호
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• pp.155-166
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• 2010

Objetives : The purpose of this study was to examine the pathway of anti-allergic effects of Aconiti Ciliare Tuber (ACT). Methods : We examined cell viability, ${\beta}$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation from RBL-2H3 cells pre-treatment by ACT of 1.0 mg/ml, 2.0 mg/ml separately. Results : We observed that ACT reduced the secretion of ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Futhermore, ACT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions : These results show that ACT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. This improves that ACT could be used as an anti-allergic medicine.

• 한국식품영양학회지
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• 제28권4호
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• pp.695-701
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• 2015

Previous reports revealed that DMfree (green tea extract) inhibited expression of the IL-6 gene in Mycobacterium lepraeinfected MSCs (mesenchymal stem cells). This study aimed to measure IL-6, $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2 production in M. leprae-infected MSCs using ELISA. To confirm the effect of DMfree on IL-6 and signal transduction, a western blotting test was performed. DMfree inhibited the expression of IL-6 in the MSCs and the heterodimer of STAT3, which also affects the expression of multiple genes. Though DMfree pre-treatment of control MSCs produced a baseline level of IL-6, it significantly inhibited the production of IL-6 in M. leprae-infected MSCs. There was no significant difference in IL-6 production between 1 and 7 day treatment groups. M. leprae-infected MSCs produced more $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2, but DMfree could not inhibit their production at a physiological concentration. This is different from other reports that used higher concentration of EGCG treatment, resulting in significant inhibition of the cytokines. The inhibition appears to be related to the concentration of EGCG. These results indicate that DMfree can alleviate inflammation involving IL-6.

• 한방재활의학과학회지
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• 제29권4호
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• pp.1-14
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• 2019

Objectives The object of this study was to assess the effect of Gyejibokryunghwan (GBH) on anti-oxidant and anti-inflammatory activities in RAW 264.7 cells and on factors associated with fracture union in tibia-fractured rats. Methods The 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity was measured to assess anti-oxidant activity. The production of nitric oxide (NO), interleukin-6 (IL-6), interleukin-$1{\beta}$ ($IL-1{\beta}$) and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to assess anti-inflammatory activity. The production of osteocalcin, calcitonin, carboxy-terminal telepeptides of type II collagen (CTXII), transforming growth factor-${\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2) in serum of tibia-fractured rats were measured to assess the effects of fracture union. X-rays were taken every two weeks from 0 to 4th week to assess fracture union effect. Results DPPH radical scavenging activity of GBH was increased according to concentration of GBH in RAW 264.7 cell. NO, prostaglandin $E_2$ ($PGE_2$), IL-6, $IL-1{\beta}$ and $TNF-{\alpha}$ were significantly decreased, indicating anti-inflammatory effect. Osteocalcin, calcitonin, $TGF-{\beta}$ were significantly increased in the experimental groups. CTXII was significantly decreased in the experimental groups. BMP-2 was not significantly changed in the experimental groups. The X-ray showed that the experimental group has better healing effects on tibia-fractured rats than control group. Conclusions From above result, GBH has an effect on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells. GBH showed significant results in factors related with fracture union and radiologic examination. In conclusion, GBH can help fracture union and it well be expected to be used actively in clinics.