• 농업생명과학연구
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• 제50권2호
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• pp.151-160
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• 2016

염증성 사이토카인은 파골세포형성과정에서 중요한 요인이며, 뼈의 흡수는 자주 골다공증과 연결된다. 설포라판은 보로콜리의 화뢰로 부터 분리된 물질로 염증성 사이토카인을 억제한다고 알려져 있다. 본 실험에서는 Receptor activator of nuclear factor kappaB ligand(RANKL)로 자극된 세포에서 설포라판이 파골세포 형성 억제에 대한 효과를 측정하였다. 설포라판은 대식세포인 RAW 264.7 세포에서 파골세포 특이 마커 유전자인 tartrate-resistant acid phosphatase(TRAP), Cathepsin K, matrix metalloproteinase 9(MMP-9), calcitonin receptor을 저해하였으며, TRAP, MMP-9, tumor necrosis factor receptor-associated factor 6(TRAF6)와 전사인자인 nuclease factor of activated T cells(NFATc1)의 단백질 발현과 RANKL로 자극하였을 때 전자인사인 nuclear factor kappaB(NF-kappaB)의 전사활성도 억제 하였다. 이와 같은 결과로 설포라판이 NF-kappaB의 전사활성 억제뿐만 아니라, 파골세포형성인자(TRAP, cathepsin K, MMP-9, calcitonin, NFATc1)와 NFATc1의 발현을 억제시키는 효과가 있음을 확인하였다.

• 대한한방부인과학회지
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• 제33권3호
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• pp.95-118
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• 2020

목 적: 이 생체 내 연구의 목적은 인도메타신과 비교했을 때 쥐의 원발성 월경통에 대한 계지 복령환 열수 추출물의 진통 및 개선 효과를 관찰하는 것이다. 방 법: 랫트 PD 모델을 만들기 위해, Estradiol benzoate를 10일간 투여한 다음 마지막 10회 estradiol benzoate 투여 1시간 후 1 U/kg의 oxytocin을 투여하였다. 계지복령환 열수 추출물을 500, 250 및 125 mg/kg 용량으로 매일 1회씩 10일간 투여하였다. 이후 체중 및 실험 기간 동안의 체중 증가량, 자궁 중량 및 육안부검 소견, 진통 활성을 나타내는 abdominal writhing test, 자궁 조직 내 지질 과산화(Malondialdehyde, MDA 함량) 및 항산화 방어 시스템 - glutathione (GSH) 함량, superoxide dismutase (SOD) 및 catalase (CAT) 활성, Nuclear factor-κB (NF-κB) 및 cyclooxygenase (COX)-2 mRNA의 발현, 자궁의 조직병리학적 변화, tumor necrosis factor (TNF)-α 및 inducible nitric oxide synthase (iNOS)로 나타나는 면역조직 화학적 변화를 관찰하였다. 계지복령환 열수 추출물의 결과는 인도메타신 투여 쥐의 결과와 비교하였다. 결 과: Estradiol benzoate 및 oxytocin 투여 결과, 현저한 체중 및 증체량, 자궁 GSH 함량, SOD와 CAT 활성의 감소와 abdominal writhing 반응, 자궁 지질 과산화(MDA 함량), 자궁 중량, NF-κB 및 COX-2 mRNA 발현의 증가가 TNF-α 및 iNOS 면역반응세포와 염증세포 침윤 증가, 자궁의 충혈 및 확대와 함께 관찰되었다. 이는 전형적인 염증 및 산화 스트레스성 원발성 월경통이 잘 유도되었음을 의미한다. 한편 이러한 소견은 계지복령환 열수 추출물의 투여에 의해 용량의존적으로 현저히 억제되었으며, 인도메타신의 억제 효과보다는 낮았다. 결 론: 이 연구에서 얻은 결과는 계지복령환 열수 추출물이 estradiol benzoate 및 oxytocin으로 유도된 원발성 월경통에 용량-의존적으로 유리한 진통 및 개선 활성을 가짐을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.589-597
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• 2015

It has been reported that overexpression of MUC5AC induced by excessive inflammation leads to airway obstruction in respiratory diseases such as chronic obstructive pulmonary disease and asthma. 15-Hydroxyeicosatetraenoic acid (15-HETE) has been reported to have anti-inflammatory effects, but the role of 15-HETE in respiratory inflammation has not been determined. Therefore, the aim of this study was to investigate the effects of 15-HETE on MUC5AC expression and related pathways. In this study, phorbol-12-myristate-13-acetate (PMA) was used to stimulate NCI-H292 bronchial epithelial cells in order to examine the effects of 15-HETE. 15-HETE inhibited PMA-induced expression of MUC5AC mRNA and secretion of MUC5AC protein. Moreover, 15-HETE regulated matrix metallopeptidase 9 (MMP-9), mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK). In addition, 15-HETE decreased the nuclear translocation of specificity protein-1 (Sp-1) transcription factor and nuclear factor κB (NF-κB). Furthermore, 15-HETE enhanced the transcriptional activity of peroxisome proliferator-activated receptor gamma (PPARγ) as a PPARγ agonist. This activity reduced the phosphorylation of protein kinase B (PΚB/Akt) by increasing the expression of phosphatase and tensin homolog (PTEN). In conclusion, 15-HETE regulated MUC5AC expression via modulating MMP-9, MEK/ERK/Sp-1, and PPARγ/PTEN/Akt signaling pathways in PMA-treated respiratory epithelial cells.

• 한국축산식품학회지
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• 제44권3호
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• pp.699-709
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• 2024

Oxya chinensis sinuosa (OC) is a well-known edible insect. Several researches on the health benefits of OC consumption have been performed to date; however, their effect on eye health remains largely unknown. This study aimed to assess the protective effects of OC extracts on the oxidative stress on the retinal pigment epithelium (RPE) cells. Oxidative damage has been identified as one of the key regulatory factors in agerelated macular degeneration. H₂O₂-induced reactive oxygen species (ROS) production, a well-known oxidative stress factor, can cause cell death in retinal pigment epithelia cells. In this study, we found that three OC extracts effectively prevented H₂O₂-induced ROS production and subsequent death of ARPE-19 cells in a dose-dependent manner. In addition, the OC extracts inhibited the phosphorylation of mitogen-activated protein kinases including p38, JNK, and ERK. The OC extracts restored IκBα degradation induced by H₂O₂, indicating that OC extracts suppressed the activation of nuclear factorκB. Furthermore, the three OC extracts were shown to have antioxidant effects by upregulating the intracellular expression of key antioxidant proteins such as SOD, NQO, and HO-1. Here we demonstrated the antioxidant and anti-apoptotic effects of the OC extracts on ARPE-19, indicating their potential role in improving eye health. These results suggest that three OC extracts plays a critical role in oxidative stress-induced cell death protects in ARPE-19 cells.

• Biomolecules & Therapeutics
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• 제30권6호
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• pp.540-545
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• 2022

Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

• Biomolecules & Therapeutics
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• 제17권3호
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• pp.219-240
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• 2009

Since NF-${\kappa}B$ has been identified as a transcription factor associated with immune cell activation, groups of researchers have dedicated to reveal detailed mechanisms of nuclear factor of ${\kappa}B$ (NF-${\kappa}B$) in inflammatory signaling for decades. The various molecular components of NF-${\kappa}B$ transcription factor pathway have been being evaluated as important therapeutic targets due to their roles in diverse human diseases including inflammation, cystic fibrosis, sepsis, rheumatoid arthritis, cancer, atherosclerosis, ischemic injury, myocardial infarction, osteoporosis, transplantation rejection, and neurodegeneration. With regards to new drugs directly or indirectly modulating the NF-${\kappa}B$ pathway, FDA recently approved a proteasome inhibitor bortezomib for the treatment of multiple myeloma. Many pharmaceutical companies have been trying to develop new drugs to inhibit various kinases in the NF-${\kappa}B$ signaling pathway for many therapeutic applications. However, a gene knock-out study for $IKK{\beta}$ in the NF-${\kappa}B$ pathway has given rise to controversies associated with efficacy as therapeutics. Mice lacking hepatocyte $IKK{\beta}$ accelerated cancer instead of preventing progress of cancer. However, it is clear that pharmacological inhibition of $IKK{\beta}$ appears to be beneficial to reduce HCC. This article will update issues of the NF-${\kappa}B$ pathway and inhibitors regulating this pathway.

• 한국버섯학회지
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• 제19권3호
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• pp.225-233
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• 2021

염증은 외부 자극으로부터 보호하는 중요한 면역반응이다(Jeong et al., 2012). 그러나 NO 및 inflammatory cytokine과 같은 염증 매개 인자의 과도한 생성은 비정상적인 염증 반응을 초래할 수 있다(Paradise et al., 2010). 염증 매개 인자의 생성과 염증 신호 전달을 조절하는 중요한 역할을 하는 대식세포는 LPS 자극에 의해 NF-κB가 활성화 되어 inflammatory cytokine 등의 염증 매개 인자들이 분비가 증가되며 염증 반응을 심화시킨다(Lee et al., 2012). 본 연구에서는 영지버섯균 발효 꾸지뽕나무 가지 톱밥 추출물을 이용하여 LPS로 염증을 유도한 Raw264.7세포에서 염증 관련 인자들의 생성 및 발현에 미치는 영향을 분석하여 항염증 효과를 확인하였다. NO는 염증에서 중요한 역할을 하며 대식세포의 염증 반응 조절 평가시 대표적인 지표로 사용되며 iNOS에 의해 생성이 유도되며 PGE₂는 염증 매개 인자로 inflammatory cytokine 생성에 관여하며 COX-2에 의해 생성이 유도된다(Paradise et al., 2010; Posadas et al., 2000). 발효추출물이 RAW264.7세포에서 세포독성 없이 NO의 생성과 PGE₂의 생성을 감소시켰으며 이 결과는 발효 추출물 처리에 의해 iNOS와 COX-2의 발현이 감소한 것과 일치하였다. 또한 염증 반응을 조절하는 대표적인 inflammatory cytokine으로 알려진 IL-1β, TNF-α의 생성이 감소되었다. 따라서 발효추출물은 NO, PGE₂, inflammatory cytokines 생성을 감소시켜 항염증 효과를 나타낸다고 볼 수 있다. NF-κB는 iNOS, COX-2 및 inflammatory cytokine의 발현을 조절하는 전사인자로 IκB와의 결합에 의해 NF-κB의 핵 내 이동이 억제되며 불활성화상태로 존재한다. LPS 자극에 의해 IκB가 인산화 및 분해되면 NF-κB가 활성화되어 핵 내로 이동하여 염증성 매개인자들의 발현을 유도하고, 염증성 질환 뿐만 아니라 다양한 질환을 유발시키는 것으로 알려져 있다 (Kawai and Akira, 2006; Ghosh and Ksarin, 2002). 본 연구에서는 LPS에 의해 자극된 RAW264.7 세포에서 IκB의 분해 및 NF-κB의 전이가 발효추출물에 의해 감소됨을 확인하였으며 발효추출물에 의한 iNOS, COX-2 및 inflammatory cytokine 감소는 NFκB 활성화 감소에 의해 조절되는 것으로 사료된다. 이러한 결과들을 통해서 발효추출물이 NF-kB 활성화 억제를 통해 염증 매개 인자들의 생성 및 발현을 감소시킴으로서 염증 반응 억제 효과를 나타냄을 확인하여, 염증 관련 질환에 대해 예방 및 개선을 위한 항염증 기능성 소재로 사용될 수 있음을 시사한다.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.385-393
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• 2008

As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we evaluated the effects of the methanol extract of Polytrichum commune Hedw (PCM) (Polytrichaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines release in murine macrophage cell line RAW 264.7. PCM potently inhibits the production of NO, $PGE_2$, tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Consistent with these results, PCM also concentration-dependently inhibited LPS-induced inducible NO synthase (iNOS) and cyclooxygase (COX)-2 at the protein levels, and iNOS, COX-2, TNF-$\alpha$ and IL-6 at the mRNA levels without an appreciable cytotoxic effect on RAW 264.7 macrophag cells. Furthermore, PCM inhibited LPS-induced nuclear factor-kappa B (NF-$\kappa$B) activation as determined by NF-$\kappa$B reporter gene assay, and this inhibition was associated with a decrease in the nuclear translocation of p65 and p50 NF-$\kappa$B. Taken together, these results suggest that PCM may play an anti-inflammatory role in LPS-stimulated RAW 264.7 macrophages through the inhibitory regulation of iNOS, COX-2, TNF-$\alpha$ and IL-6 via NF-$\kappa$B inactivation.

• Journal of Microbiology and Biotechnology
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• 제30권6호
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• pp.822-829
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• 2020

Nutraceutical treatments can reduce inflammation and prevent the development of inflammatory diseases. In this study, the anti-inflammatory effects of Smilax guianensis Vitman extract (SGE) were examined. SGE suppressed lipopolysaccharide (LPS)-mediated nitrite production in RAW 264.7 cells. SGE also prevented the LPS-induced expression of inducible nitric oxide synthase (iNOS) but not cyclooxygenase (COX)-2. Western blot analysis showed that SGE attenuated LPS-induced phosphorylation of IκB kinase (IKK), inhibitor of kappa B (IκB), and p65. Additionally, SGE inhibited LPS-induced IκB degradation in RAW 264.7 cells. Western blot analysis of the cytosolic and nuclear fractions, as well as immunofluorescence assay results, revealed that SGE suppressed LPS-induced p65 nuclear translocation in RAW 264.7 cells. Moreover, SGE reduced LPS-induced interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) mRNA expression and IL-1β and IL-6 protein expression in RAW 264.7 cells. Collectively, these results indicate that SGE suppresses the NF-κB signaling pathway and thereby inhibits the production of NO, IL-1β, and IL-6.

• Biomolecules & Therapeutics
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• 제25권4호
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• pp.383-389
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• 2017

Dexmedetomidine is an ${\alpha}2$-adrenergic receptor agonist that exhibits a protective effect on ischemia-reperfusion injury of the heart, kidney, and other organs. In the present study, we examined the neuroprotective action and potential mechanisms of dexmedetomidine against ischemia-reperfusion induced cerebral injury. Transient focal cerebral ischemia-reperfusion injury was induced in Sprague-Dawley rats by middle cerebral artery occlusion. After the ischemic insult, animals then received intravenous dexmedetomidine of $1{\mu}g/kg$ load dose, followed by $0.05{\mu}g/kg/min$ infusion for 2 h. After 24 h of reperfusion, neurological function, brain edema, and the morphology of the hippocampal CA1 region were evaluated. The levels and mRNA expressions of interleukin-$1{\beta}$, interleukin-6 and tumor nevrosis factor-${\alpha}$ as well as the protein expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-${\kappa}Bp65$, inhibitor of ${\kappa}B{\alpha}$ and phosphorylated of ${\kappa}B{\alpha}$ in hippocampus were assessed. We found that dexmedetomidine reduced focal cerebral ischemia-reperfusion injury in rats by inhibiting the expression and release of inflammatory cytokines and mediators. Inhibition of the nuclear factor-${\kappa}B$ pathway may be a mechanism underlying the neuroprotective action of dexmedetomidine against focal cerebral I/R injury.