• 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.

• Journal of Nutrition and Health
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• 제49권6호
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• pp.420-428
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• 2016

본 연구에서는 Sprague-Dawley 종 흰 쥐 수컷을 정상 대조군 (C), 알코올 군 (E), 알코올 + 100 mg/kgBW 곤드레 에탄올 추출물군 (E+LCS), 알코올 + 500 mg/kgBW 곤드레 에탄올 추출물군 (E+HCS)으로 나누어 Lieber-DeCarli control diet 혹은 Lieber-DeCarli ethanol diet를 8주간 공급하였으며, 이때 곤드레 에탄올 추출물은 액상 사료에 직접 섞어 공급하였다. 알코올과 곤드레 에탄올 추출물의 식이 공급 종료 후 간 조직의 지방구 축적 정도를 살펴본 결과, E+LCS군과 E+HSC군은 E군에 비해 지방간 발생이 유의적으로 억제되었으며, 정상 대조군인 C군과 유의적인 차이가 없었다. 이와 비슷하게, 곤드레 에탄올 추출물의 공급은 알코올에 의해 증가된 간 조직과 혈청의 중성지방 농도를 유의적으로 낮추었으며, 혈청 AST와 혈청 ALT 활성도 정상 대조군 수준으로 회복시키는 것으로 나타났다. 한편, 곤드레 에탄올 추출물의 공급은 p-AMPK과 p-ACC 단백질 수준을 농도 의존적으로 증가시켰으며, 두 단백질 모두 E군에 비해 E+HSC군에서 유의적으로 높게 나타났다. 또한 FAS mRNA와 SCD1 mRNA 수준은 E군에 비해 E+HSC군에서 유의적으로 낮게 나타났다. 곤드레 에탄올 추출물은 간 조직에서 알코올 공급에 의해 증가된 $NF{\kappa}B$의 활성을 유의적으로 낮추었으며, $NF{\kappa}B$의 표적 단백질인 $TNF{\alpha}$ 단백질 수준을 농도의존적으로 낮추었다. 본 연구 결과를 통해 곤드레는 알코올에 의한 지방간 발생 및 관련된 간 손상을 유의적으로 억제할 수 있음을 확인하였으며 이 과정에서 AMPK 활성 증가와 $NF{\kappa}B$ 활성 억제가 관여함을 제시하였다.

• 한국식품영양과학회지
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• 제45권2호
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• pp.194-201
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• 2016

본 연구에서는 lipopolysaccharide(LPS)로 자극한 마우스 대식세포인 RAW 264.7 세포에서 가락진두발 에탄올 추출물(CNYEE)의 항염증 효과를 알아보기 위하여 nitric oxide(NO)와 pro-inflammatory cytokine의 분비량을 확인하였다. 그 결과 CNYEE 모든 농도에서 LPS만을 처리한 대조군과 비교하였을 때 유의성 있게 NO와 pro-inflammatory cytokine의 분비량을 저해하였으며, 특히 $100{\mu}g/mL$ 농도에서는 IL-6의 분비량을 70% 이상 억제하였고, TNF-${\alpha}$ 및 IL-$1{\beta}$의 분비량은 50% 이상의 억제 효과를 나타내었다. CNYEE에 의한 염증매개물질의 분비 감소가 전사인자인 nuclear factor-${\kappa}B$(NF-${\kappa}B$)의 핵 내 전이 pathway를 저해함으로써 나타난 결과인지 확인하기 위하여 iNOS, COX-2 및 NF-${\kappa}B$ p65의 단백질 발현량을 관찰한 결과, 비교적 낮은 농도인 $50{\mu}g/mL$에서 40% 이상의 저해능을 보인 것으로 보아 NO와 cytokine의 분비 억제 결과가 NF-${\kappa}B$ pathway를 저해함으로써 나타난 것임을 유추할 수 있었다. 또한 LPS에 의해 증가한 mitogen-activated protein kinases의 인산화를 확인한 결과, CNYEE 처리에 의해 농도 의존적으로 유의성 있게 저해되었다. 이러한 결과를 종합해볼 때 가락진두발 에탄올 추출물은 염증매개물질의 분비를 효과적으로 저해함으로써 추후 천연물로서 염증 치료제의 개발이 가능할 것으로 생각한다.

• 생명과학회지
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• 제21권5호
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• pp.656-663
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• 2011

Licochalcone은 감초의 주요 생리활성 물질로 항균작용, 항암작용 등의 다양한 효과가 있는 것으로 알려져 있다. 최근 감초로부터 licochalcone E가 분리, 동정 되었을 뿐만 아니라, 효과적인 licochalcone E의 합성을 위해 다양한 합성법이 개발되고 있다. 반면, licochalcone E의 생리활성 연구는 매우 미비한 상태이다. 본 연구는 licochclcone E의 항염증 활성과 그 기전의 일단을 밝히는 것을 목적으로 진행 되었다. 생쥐의 대식세포주인 RAW264.7 세포에 lipopolysaccharide (LPS)를 처리 염증반응을 유도하고, licochalcone E를 처리한 한 결과, licochalconeE는 LPS 처리에 의한 nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) 및 염증성 사이토카인의 분비를 현저히 억제시키는 것을 관찰 할 수 있었으며, NO와 $PGE_2$ 생합성효소인 iNOS와 COX-2 단백질의 발현 또한 억제시킴을 확인할 수 있었다. 이러한 licochalcone E의 항염증 활성의 기전을 밝히기 위해 염증반응에 핵심적인 역할을 하는 전사인자인 nuclear factor-${\kappa}$B (NF-${\kappa}$B)의 활성을 관찰한 결과, licochalcone E의 처리가 NF-${\kappa}$B의 DNA결합을 억제하는 것을 확인 하였다. 이러한 연구결과로 볼 때 licochalcone E가 NF-${\kappa}$B의 활성을 억제하여, 염증반응의 매개물질인 NO, $PGE_2$, 염증성 사이토카인 등의 생성을 억제함으로 항염증활성을 나타내는 것으로 판단된다.

• Biomolecules & Therapeutics
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• 제24권5호
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• pp.543-551
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• 2016

This study was designed to evaluate the pharmacological effects of Vaccinium bracteatum Thunb. methanol extract (VBME) on microglial activation and to identify the underlying mechanisms of action of these effects. The anti-inflammatory properties of VBME were studied using lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. We measured the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin $E_2$ ($PGE_2$), tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1 beta (IL-$1{\beta}$), and interleukin-6 (IL-6) as inflammatory parameters. We also examined the effect of VBME on intracellular reactive oxygen species (ROS) production and the activity of nuclear factor-kappa B p65 (NF-${\kappa}B$ p65). VBME significantly inhibited LPS-induced production of NO and $PGE_2$ and LPS-mediated upregulation of iNOS and COX-2 expression in a dose-dependent manner; importantly, VBME was not cytotoxic. VBME also significantly reduced the generation of the pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. In addition, VBME significantly dampened intracellular ROS production and suppressed NF-${\kappa}B$ p65 translocation by blocking $I{\kappa}B-{\alpha}$ phosphorylation and degradation in LPS-stimulated BV2 cells. Our findings indicate that VBME inhibits the production of inflammatory mediators in BV-2 microglial cells by suppressing NF-${\kappa}B$ signaling. Thus, VBME may be useful in the treatment of neurodegenerative diseases due to its ability to inhibit inflammatory mediator production in activated BV-2 microglial cells.

• Journal of Ginseng Research
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• 제42권4호
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• pp.496-503
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• 2018

Background: Korean ginseng (Panax ginseng) plays an anti-inflammatory role in a variety of inflammatory diseases such as gastritis, hepatitis, and colitis. However, inflammation-regulatory activity of the calyx of the P. ginseng berry has not been thoroughly evaluated. To understand whether the calyx portion of the P. ginseng berry is able to ameliorate inflammatory processes, an ethanolic extract of P. ginseng berry calyx (Pg-C-EE) was prepared, and lipopolysaccharide-activated macrophages and HEK293 cells transfected with inflammation-regulatory proteins were used to test the anti-inflammatory action of Pg-C-EE. Methods: The ginsenoside contents of Pg-C-EE were analyzed by HPLC. Suppressive activity of Pg-C-EE on NO production, inflammatory gene expression, transcriptional activation, and inflammation signaling events were examined using the Griess assay, reverse transcription-polymerization chain reaction, luciferase activity reporter gene assay, and immunoblotting analysis. Results: Pg-C-EE reduced NO production and diminished mRNA expression of inflammatory genes such as cyclooxygenase-2, inducible NO synthase, and tumor necrosis factor-${\alpha}$ in a dose-dependent manner. This extract suppressed luciferase activity induced only by nuclear factor-${\kappa}B$. Interestingly, immunoblotting analysis results demonstrated that Pg-C-EE reduced the activities of protein kinase B (AKT)1 and AKT2. Conclusion: These results suggest that Pg-C-EE may have nuclear-factor-${\kappa}B$-targeted anti-inflammatory properties through suppression of AKT. The calyx of the P. ginseng berry is an underused part of the ginseng plant, and development of calyx-derived extracts may be useful for treatment of inflammatory diseases.

• 한방재활의학과학회지
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• 제29권2호
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• pp.135-147
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• 2019

Objectives This study was performed to evaluate the effect of Pyrola japonica extract (NJ) and its principal constituent, homoarbutin (HA) on osteoclast differentiation and gene expression and bone resorption. The osteoclastogenesis and gene expression were determined in receptor activator of nuclear factor kappa B ligand (RANKL)-stimulated RAW264.7 cell. Methods In order to evaluate the effect of HA extracted from NJ on bone resorption, osteoclasts were used to be differentiated and formed by stimulating RAW264.7 cells with RANKL. Tartarate-resistant acid phosphatase (TRAP) (+) polynuclear osteoclast formation ability was evaluated, and differentiation control genes including cathepsin K, matrix metalloproteinases-9 (MMP-9), and TRAP in osteoclast differentiation were analyzed by real-time polymerase chain reaction (PCR). Immunoblotting was performed to measure the effect of mitogen-activated protein kinase (MAPK) factors on bone resorption, and the effect of osteoclasts on osteoclast differentiation was measured. Results Both NJ and high concentration of HA blocked RANKL-stimulated differentiation from RAW264.7 cell to TRAP-positive multinucleated cells. NJ reduced RANKL-induced expression of TRAP, cathepsin K. Both NJ and high concentration of HA inhibited RANKL-mediated expression of MMP-9, nuclear factor of activated T-cells, cytoplasmic 1, and cellular Jun-fos. NJ suppressed RANKL-stimulated expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase, tumor necrosis factor-alpha, and levels of interleukins. Both NJ and HA decreased bone resorption in osteoclast-induced bone pit formation model. Conclusions These results suggest that NJ and HA blocked bone resorption by decreasing RANKL-mediated osteoclastogenesis through down-regulation of genes for osteoclast differentiation.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.11-18
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• 1999

Nuclear factor $_{\kappa}B\;(NF-_{\kappa}B)$ activation is modulated by various protein kinases. Activation of $NF-_{\kappa}B$ is known to be important in the regulation of cell viability. The present study investigated the effect of inhibitors of protein tyrosine kinase (PTK), protein kinase C (PKC) and protein kinase A (PKA) on $NF-_{\kappa}B$ activity and the viability of bovine cerebral endothelial cells (BCECs). In serum-deprivation-induced BCEC death, low doses of $TNF{\alpha}$ showed a protective effect. $TNF{\alpha}$ induced $NF-_{\kappa}B$ activation within 4 h in serum-deprivation. PTK inhibitors (herbimycin A and genistein) and PKC inhibitor (calphostin C) prevented $NF-_{\kappa}B$ activation stimulated by $TNF{\alpha}.$ Likewise, these inhibitors prevented the protective effect of $TNF{\alpha}.$ In contrast to $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activity, basal $NF-_{\kappa}B$ activity of BCECs in media containing serum was suppressed only by calphostin C, but not by herbimycin A. As well BCEC death was also induced only by calphostin C in serum-condition. H 89, a PKA inhibitor, did not affect the basal and $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activities and the protective effect of $TNF{\alpha}$ on cell death. These data suggest that modulation of $NF-_{\kappa}B$ activation could be a possible mechanism for regulating cell viability by protein kinases in BCECs.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.637-642
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• 2021

In this study, we investigated whether eriodictyol exerts an effect on the production and gene expression of MUC5AC mucin in human pulmonary epithelial NCI-H292 cells. The cells were pretreated with eriodictyol for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h. The effect of eriodictyol on PMA-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated. Eriodictyol suppressed the MUC5AC mucin production and gene expression induced by PMA via suppression of inhibitory kappa Bα degradation and NF-κB p65 nuclear translocation. These results suggest that eriodictyol inhibits mucin gene expression and production in human airway epithelial cells via regulation of the NF-κB signaling pathway.

• Biomolecules & Therapeutics
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• 제30권5호
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• pp.473-478
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• 2022

In this study, we examined whether engeletin exerts an effect on the gene expression of MUC5AC mucin, in human pulmonary epithelial NCI-H292 cells. The cells were pretreated with engeletin for 30 min and stimulated with phorbol 12-myristate 13-acetate (PMA), for the following 24 h. The effect of engeletin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated. Engeletin suppressed the mRNA expression and production of MUC5AC mucin, induced by PMA through the inhibition of degradation of inhibitory kappa Bα (IkBα) and NF-kB p65 nuclear translocation. These results suggest engeletin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.