• 생명과학회지
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• 제22권9호
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• pp.1137-1144
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• 2012

염증반응은 유해한 물질이나 병원체에 대항하여 활성화되는 생체 방어 기전이다. 그러나 과도한 염증반응은 그 자체가 생체에 좋지 않은 영향을 미칠 수 있다. 대식세포는 지질다당류와 같은 병원체를 인식한 후, NF-${\kappa}B$ 경로의 활성화를 포함한 다양한 경로를 통하여 산화질소와 같은 염증매개인자들을 분비하는 면역세포이다. 본 연구에서는 지질다당류로 활성화시킨 RAW 264.7 대식세포를 이용하여 등골나물(Eupatorium chinensis var. simplicifolium) 뿌리, 줄기 그리고 꽃 추출물들의 항염증 효과를 알아보았다. 그 중 등골나물 뿌리의 추출물은 농도의존적으로 산화질소의 생성을 감소시켰으며, 산화질소 합성유도효소(inducible nitric oxide synthase)와 고리형 산소화효소-2(cyclooxygenase-2)의 발현을 통계적으로 유의하게 감소시켰다. 또한 등골나물 뿌리의 추출물은 NF-${\kappa}B$ 경로에 있는 MAP (mitogen activated protein) 인산화효소와 단백질 인산화효소 B (protein kinase B)의 활성화를 감소시켰으며, 억제적 kappa B (inhibitory kappa B)의 분해 또한 감소시키는 것을 관찰하였다. 이러한 결과는 등골나물 뿌리의 추출물이 NF-${\kappa}B$ 경로와 산화질소 합성유도효소 발현의 억제를 통하여 항염증작용을 나타낼 수 있음을 제시한다.

• 생명과학회지
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• 제32권2호
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• pp.125-134
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• 2022

본 연구는 노니(Morinda citrifolia)에 함유된 주요 생리활성물질이 RAW 264.7 세포에서 JAK/STAT 신호전달 경로를 통하여 항염증 작용에 관여하는 것을 조사하였다. 건조된 M. citrifolia 열매의 MeOH 추출에 의한 유기용매의 순차 분획에서 얻은 EtOAc 분획물(Mc-EtOAc)에서 가장 높은 항산화 활성을 확인하였고, H₂O₂로 유도된 RAW 264.7 세포의 산화적 스트레스에 대한 세포보호 효과는 처리 농도의존적으로 세포독성을 차단하였다. LPS 처리로 71.6%의 RAW 264.7 세포 생존율에서 240 ㎍/ml의 Mc-EtOAc를 전처리한 군에서 84.5%의 세포 생존율 증가를 보였다. LPS로 유도된 RAW 264.7 세포의 NO 생성 저해활성은 240 ㎍/ml의 Mc-EtOAc에서 양성 대조군의 절반 수준으로 NO의 생성양을 감소시켰다. Mc-EtOAc 처리로 iNOS의 발현량은 농도의존적으로 유의하게 감소하였고, COX-2의 발현은 LPS 유도로 3배 정도로 증가되었으나, 120, 240 ㎍/ml의 Mc-EtOAc를 전처리시 농도의존적으로 유의하게 감소시켰다. 또한 pro-inflammatory cytokine IL-1β와 TNF-α의 mRNA 발현도 억제하였다. 이러한 Mc-EtOAc의 항염증 작용이 상위 신호전달 경로인 JAK/STAT 신호전달 경로 통해 일어나지를 조사한 결과, Mc-EtOAc의 전처리로 LPS로 유도된 RAW 264.7 세포에서 pJAK1과 pSTAT3의 인산화 정도가 유의성 있게 감소하였다. 따라서 RAW 264.7 세포에서 Mc-EtOAc의 항염증 효과는 JAK1/STAT3 신호전달 경로를 통해 염증반응을 억제하는 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제23권3호
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• pp.238-244
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• 2015

Macrophage-derived chemokine, C-C motif chemokine 22 (MDC/CCL22), is one of the inflammatory chemokines that controls the movement of monocytes, monocyte-derived dendritic cells, and natural killer cells. Serum and skin MDC/CCL22 levels are elevated in atopic dermatitis, which suggests that the chemokines produced from keratinocytes are responsible for attracting inflammatory lymphocytes to the skin. A major signaling pathway in the interferon-${\gamma}$ (IFN-${\gamma}$)-stimulated inflammation response involves the signal transducers and activators of transcription 1 (STAT1). In the present study, we investigated the anti-inflammatory effect of dieckol and its possible action mechanisms in the category of skin inflammation including atopic dermatitis. Dieckol inhibited MDC/CCL22 production induced by IFN-${\gamma}$ (10 ng/mL) in a dose dependent manner. Dieckol (5 and $10{\mu}M$) suppressed the phosphorylation and the nuclear translocation of STAT1. These results suggest that dieckol exhibits anti-inflammatory effect via the down-regulation of STAT1 activation.

• Biomolecules & Therapeutics
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• 제23권1호
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• pp.39-44
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• 2015

Tolfenamic acid (TA) is a traditional non-steroid anti-inflammatory drug (NSAID) and has been broadly used for the treatment of migraines. Nuclear factor kappa B (NF-${\kappa}B$) is a sequence-specific transcription factor and plays a key role in the development and progression of inflammation and cancer. We performed the current study to investigate the underlying mechanisms by which TA suppresses inflammation focusing on NF-${\kappa}B$ pathway in TNF-${\alpha}$ stimulated human normal and cancer cell lines and lipopolysaccharide (LPS)-stimulated mouse macrophages. Different types of human cells (HCT116, HT-29 and HEK293) and mouse macrophages (RAW264.7) were pre-treated with different concentrations of TA and then exposed to inflammatory stimuli such as TNF-${\alpha}$ and LPS. Transcriptional activity of NF-${\kappa}B$, $l{\kappa}B-{\alpha}$-degradation, p65 translocation and mitogen-activated protein kinase (MAPK) activations were measured using luciferase assay and Western blots. Pre-treatment of TA repressed TNF-${\alpha}$- or LPS-stimulated NF-${\kappa}B$ transactivation in a dose-dependent manner. TA treatment reduced degradation of $l{\kappa}B-{\alpha}$ and subsequent translocation of p65 into nucleus. TA significantly down-regulated the phosphorylation of c-Jun N-terminal kinase (JNK). However, TA had no effect on NF-${\kappa}B$ signaling and JNK phosphorylation in HT-29 human colorectal cancer cells. TA possesses anti-inflammatory activities through suppression of JNK/NF-${\kappa}B$ pathway in different types of cells.

• Biomolecules & Therapeutics
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• 제21권5호
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• pp.332-337
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• 2013

Microglial activation plays an important role in the development and progression of various neurological disorders such as cerebral ischemia, multiple sclerosis, and Alzheimer's disease. Thus, controlling microglial activation can serve as a promising therapeutic strategy for such brain diseases. In the present study, we showed that kalopanaxsaponin A, a triterpenoid saponin isolated from Kalopanax pictus, inhibited inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor (TNF)-${\alpha}$ expression in lipopolysaccharide (LPS)-stimulated microglia, while kalopanaxsaponin A increased anti-inflammatory cytokine interleukin (IL)-10 expression. Subsequent mechanistic studies revealed that kalopanaxsaponin A inhibited LPS-induced DNA binding activities of NF-${\kappa}B$ and AP-1, and the phosphorylation of JNK without affecting other MAP kinases. Furthermore, kalopanaxsaponin A inhibited the intracellular ROS production with upregulation of anti-inflammatory hemeoxygenase-1 (HO-1) expression. Based on the previous reports that JNK pathway is largely involved in iNOS and proinflammatory cytokine gene expression via modulating NF-${\kappa}B$/AP-1 and ROS, our data collectively suggest that inhibition of JNK pathway plays a key role in anti-inflammatory effects of kalopanaxsaponin A in LPS-stimulated microglia.

• Natural Product Sciences
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• 제23권3호
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• pp.183-191
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• 2017

Luteolin 5-O-glucoside is the major flavonoid from Korean thistle, Cirsium maackii. We previously reported the anti-inflammatory activities of luteolin 5-O-glucoside in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In this study, we determined the anti-inflammatory mechanisms of luteolin 5-O-glucoside through the inhibition of nitric oxide (NO) production in vitro and in vivo. Results revealed that luteolin 5-O-glucoside dose-dependently inhibited NO production and expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells. Luteolin 5-O-glucoside also significantly inhibited the translocation of $NF-{\kappa}B$, the activation of MAPKs, and ROS generation in LPS-induced RAW 264.7 cells. In addition, protein expressions of Nrf-2 and HO-1 were also upregulated by luteolin 5-O-glucoside treatment. Moreover, luteolin 5-O-glucoside inhibited ${\lambda}-carrageenan-induced$ mouse paw edema by 65.34% and 48.31% at doses of 50 and 100 mg/kg body weight, respectively. These findings indicate potential anti-inflammatory effect of luteolin 5-O-glucoside particularly by downregulating $NF-{\kappa}B$ and upregulating HO-1/Nrf-2 pathway.

• Mycobiology
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• 제43권4호
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• pp.450-457
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• 2015

Medicinal mushrooms have been used worldwide to treat cancer and modulate the immune system. Over the last several years, there has been increasing interest in isolating bioactive compounds from medicinal mushrooms and evaluating their health beneficial effects. Fomes fomentarius is used in traditional oriental medicine and is known to possess antioxidant, antiinflammatory, antidiabetic, and antitumor effects. In the present study, we isolated fomentariol from Fomes fomentarius and investigated its anti-inflammatory effect in murine macrophages (RAW264.7 cells) stimulated with lipopolysaccharides. Fomentariol inhibited the production of nitric oxide and intracellular reactive oxygen species triggered by lipopolysaccharides. Interestingly, fomentariol differentially regulated cytokine production triggered by lipopolysaccharides. Fomentariol effectively suppressed the production of interleukin-$1{\beta}$ and interleukin-6 but not tumor necrosis factor-${\alpha}$. The inhibitory effect of fomentariol against nitric oxide, interleukin-$1{\beta}$, and interleukin-6 production was possibly mediated by downregulation of the extracellular signal-regulated kinase signaling pathway. Taken together, our results suggest that fomentariol differentially modulated inflammatory responses triggered by lipopolysaccharides in macrophages and is one of the bioactive compounds that mediate the physiological effects of Fomes fomentarius.

• The Korean Journal of Pain
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• 제34권1호
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• pp.47-57
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• 2021

Background: Lumbar disc herniation (LDH) is a common cause of radicular pain, but the mechanism is not clear. In this study, we investigated the engagement of toll-like receptor 4 (TLR4) and the nuclear factor-kappa B (NF-κB) in radicular pain and its possible mechanisms. Methods: An LDH model was induced by autologous nucleus pulposus (NP) implantation, which was obtained from coccygeal vertebra, then relocated in the lumbar 4/5 spinal nerve roots of rats. Mechanical and thermal pain behaviors were assessed by using von Frey filaments and hotplate test respectively. The protein level of TLR4 and phosphorylated-p65 (p-p65) was evaluated by western blotting analysis and immunofluorescence staining. Spinal microglia activation was evaluated by immunofluorescence staining of specific relevant markers. The expression of proand anti-inflammatory cytokines in the spinal dorsal horn was measured by enzyme linked immunosorbent assay. Results: Spinal expression of TLR4 and p-NF-κB (p-p65) was significantly increased after NP implantation, lasting up to 14 days. TLR4 was mainly expressed in spinal microglia, but not astrocytes or neurons. TLR4 antagonist TAK242 decreased spinal expression of p-p65. TAK242 or NF-κB inhibitor pyrrolidinedithiocarbamic acid alleviated mechanical and thermal pain behaviors, inhibited spinal microglia activation, moderated spinal inflammatory response manifested by decreasing interleukin (IL)-1β, IL-6, tumor necrosis factor-α expression and increasing IL-10 expression in the spinal dorsal horn. Conclusions: The study revealed that TLR4/NF-κB pathway participated in radicular pain by encouraging spinal microglia activation and inflammatory response.

• 대한본초학회지
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• 제24권4호
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• pp.181-188
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• 2009

Objectives : In this study, we investigated the effect of methyl gallate of Paeonia suffruticosa(Moutan Cortex Radicis) on inflammatory response in activated macrophages. Methods : RAW264.7 cells were incubated with different concentrations of methyl gallate of Paeonia suffruticosa for 30 min and then stimulated with or without LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and inflammatory cytokines (TNF-$\alpha$, IL-6) were measured in culture medium by Griess assay, enzyme-immuno assay, and ELISA, respectively. The expressions of iNOS, COX-2 and cytokine mRNA and protein were determined by RT-PCR and Western blot, respectively. The $I{\kappa}-B{\alpha}$ degradation in cytosol and NF-${\kappa}B$ p65 translocation into nuclear of the cells were determined by Western blot. Results : Methyl gallate was significantly inhibited LPS-induced production of NO and PGE2 in RAW264.7 cells. Methyl gallate was also suppressed LPS-induced expression of iNOS and COX-2 mRNA and protein in the cells. Methyl gallate was inhibited LPS-induced production of TNF-$\alpha$ and IL-6 via suppression of their mRNA expressions. Methyl gallate blocked the NF-${\kappa}B$ pathway in LPS-stimulated RAW264.7 cells. Conclusions : This study suggests that methyl gallate of Paeonia suffruticosa may have an antiinflammatory property through suppressing inflammatory mediator production in activated macrophages.

• Biomolecules & Therapeutics
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• 제18권3호
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• pp.308-315
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• 2010

Essential oil-excluded Artemisia princeps Pamp var Ssajuarissuk (AP) was fermented with Lactobacillus brevis K-1, which was isolated from cabbage Kimchi, and the anti-inflammatory effects of AP and fermented AP (FAP) on lipopolysaccharide (LPS)-induced inflammatory response in peritoneal macrophages were investigated. AP and FAP inhibited LPS-induced TNF-$\alpha$, IL-$1{\beta}$, COX-2, iNOS and COX-2 expression, as well as NF-${\kappa}B$ activation. AP and FAP also reduced ear thickness, inflammatory cytokine (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced dermatitis in mice. Furthermore, AP and FAP also reduced exudate volume, cell number, protein amount, inflammatory cytokines (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation in carrageenan-induced air pouch inflammation in mice. The inhibitory effects of FAP were more potent than those of non-fermented AP. Based on these findings, we propose that FAP can improve inflammatory diseases, such as dermatitis, by inhibiting the NF-${\kappa}B$ pathway.