• Tuberculosis and Respiratory Diseases
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• 제49권3호
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• pp.332-342
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• 2000

연구배경 : 염증매개 사이토카인은 염증성 폐질환의 중요한 매개물질이다. 폐 상피세포는 염증세포에서 분비되는 사이토카인에 의해 interleukin, chemokines, colony stimulating factors와 growth factor등을 생산 및 분비함으로써 국소 염증 부위에서의 사이토카인 network에 중요한 역할을 한다. 따라서 폐 상피세포에서 염증매개 사이토카인의 발현 기전에 대한 이해는 염증성 폐질환의 기전규명과 이에 기초한 새로운 치료법의 개발에 생각된다. 대부분의 사이토카인은 NF-${\kappa}B$전사인자에 의해 발현되는데 폐 상피세포에서 염증매개 사이토키인의 발현과 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성에 관한 연구는 부족한 실정이다. 방법 : BEAS-2B, A549, NCI-H157, NCI-H719 세포에서 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 IL-8과 TNF-$\alpha$ mRNA의 발현 양상을 평가하였고 이들의 발현과 관찰하였고 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성을 평가하기 위하여 IL-l$\beta$와 TNF-$\alpha$ 자극에 의한 NF-${\kappa}B$의 활성화 및 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$의 분해 양상을 관찰하였다. 폐 상피세포의 종류에 따른 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 기전을 규명하고자 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 $I{\kappa}B{\alpha}$의 인산화와 기저상태에서 IKK$\alpha$의 발현을 평가하였다. 결과 : BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$가 분해되었고 NF-${\kappa}B$의 활성화가 관찰되었으며 IL-8과 TNF-$\alpha$mRNA의 발현이 유도되었다. NCI-H719 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B$ 분해에 의한 NF-${\kappa}B$의 활성화 및 염증매개 사이토카인의 발현이 관찰되지 않았다. BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 ${\kappa}B$의 인산화가 관찰되었지만 NCI-H719 세포에서는 관찰되지 않았다. 기저상태의 IKK$\alpha$ 발현은 세포간에 차이가 관찰되지 않았다. 결론 : 폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로는 염증매개 사이토카인 발현에 매우 중요한 역할을 하고, 일부 세포에서는 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 차이를 보이는데 이는 IKK보다 상위 단계의 세포내 신호전달체계의 이상에 기인한 것으로 생각된다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.92.1-92.1
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• 2003

In our search for NF-kB inhibitors from natural resources, we have previously identified two structurally related dilignans, manassantin A and B as specific inhibitors of NF-kB activation from Saururus chinensis. However, their molecular mechanism of action remains unclear. We here demonstrate that manassantin A and B are potent inhibitors of NF-kB activation by the suppression of transciptional activity of RelA/p65 subunit of NF-kB. These compounds significantly inhibited the induced expression of NF-kB reporter gene by LPS or TNF-a in a dose-dependent manner. (omitted)

• Tuberculosis and Respiratory Diseases
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• 제65권6호
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• pp.476-486
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• 2008

연구배경: 정상세포는 보호되고 종양세포에 독성을 보인다고 알려진 TNF유전자족으로 새로이 확인된 TRAIL이 폐암세포에서 보이는 아포프토시스 효과를 확인하고, 아포프토시스로부터 세포를 보호하는 전사인자 $NF-{\kappa}B$가 TRAIL에 의하여 활성화 되는 정도를 평가하여 MG132의 $NF-{\kappa}B$활성억제가 TRAIL 유도성 아포프토시스를 감작시키는지를 확인하기 위하여 본 연구를 시행하였다. 방법: A549(wt p53) 및 NCI-H1299(null p53) 폐암세포주를 사용하였다. 세포독성 검사는 MTT assay를 이용하였고 아포프토시스는 Annexin V assay와 FACS 분석을 이용하였다. $NF-{\kappa}B$ 전사활성은 luciferase reporter gene assay를 이용하였고 $I{\kappa}B{\alpha}$ 분해는 western blot을 이용하였으며, TRAIL에 의해 활성화된 $NF-{\kappa}B$와 DNA 결합은 electromobility shift assay와 anti-p65 antibody를 이용한 supershift assay로 확인하였다. 결과: 1) TRAIL 100 ng/ml 농도에서 wild-type p53인 A549 폐암세포는 34.4%, p53 null인 NCI-H1299 폐암세포는 26.4%의 세포사를 관찰하였다. 2) Luciferase reporter gene assay로서 TRAIL에 의한 $NF-{\kappa}B$의 활성이 A549 $IgG{\kappa}B-luc$세포에서 2.45배 증가하고 NCI-H1299 $IgG{\kappa}B-luc$세포에서는 1.47배 증가함을 관찰하여 TRAIL에 의하여 $NF-{\kappa}B$가 활성화됨을 확인하였다. 3) MG132의 전처치로 TRAIL에 의한 $NF-{\kappa}B$의 활성이 A549 세포와 NCI-H1299 세포에서 각각 기저수준의 0.24, 0.21배로 강력히 억제되었다. 4) TRAIL단독으로 30% 전후의 세포독성이 MG132 전처치 후 TRAIL을 투여하면 두 세포주 모두에서 80% 이상의 세포독성이 관찰되어 MG132가 TRAIL유도성 아포프토시스에 감작효과가 있음을 확인하였다. 결론: 이상의 결과로 TRAIL에 상대적인 내성을 보이는 폐암세포주에서 MG132가 $NF-{\kappa}B$ 활성억제로서 TRAIL유도성 아포프토시스를 강화시키는 효과가 있음을 확인할 수 있었다. 따라서 본 연구는 향후 폐암치료에 있어서 TRAIL유도성 아포프토시스가 이용될 수 있는 가능성을 확인한 기초자료가 된다고 생각된다.

• 생약학회지
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• 제35권3호통권138호
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• pp.244-249
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• 2004

The effects of hesperetin and naringenin on $NF-{\kappa}B$ activation were investigated in normal rat kidney cells transformed by temperature sensitive Rous Sarcoma Virus (tsNRK). The flavonoids, naringenin and hesperetin, significantly reduced v-Src-induced $NF-{\kappa}B$ activation as well as phosphorylation of Akt and GSK-3 in tsNRK cells, whereas these compounds did not effect on platelet-derived growth factor (PDGF)-induced $NF-{\kappa}B$ activation in $NIH3T3{\gamma}l$ cells. In addition, the DNA binding activity of SP-I was also reduced but that of AP-1 was not affected by the compounds. Our study suggests that Src-induced $NF-{\kappa}B$ activation could occur via Akt-GSK-3 pathway without $IkB{\alpha}$ degradation and that naringenin and hesperetin could be used in the treatment of cancer through the inhibition of $NF-{\kappa}B$ 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.

• 한방안이비인후피부과학회지
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• 제29권2호
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• pp.56-64
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• 2016

Objectives : The purpose of this study is to observe the effects of Gardeniae Fuctus(GF) on the metabolic process of antioxidant and anti-inflammation. Methods : 4-HNE was injected into PC-12 cell to cause oxidative stress-induced inflammatory response, and then a western blot was conducted to observe the expression of Nuclear Factor-kB (NF-kB) and c-Jun N-terminal kinase (JNK) protein that are important factors involved with inflammation. Results : 1. The Gardeniae Fuctus water extract 50 ㎍ and 100 ㎍ significantly suppressed the increase in JNK protein expression in PC-12 cell. 2. The Gardeniae Fuctus water extract 50 ㎍, 100 ㎍ and 200 ㎍ significantly suppressed the increase in NF-kB protein expression in PC-12 cell. Conclusion : These results suggest that the Gardeniae Fuctus water extract has antioxidative and anti-inflammatory activity through suppressing the activity of JNK and NF-kB.

• Journal of Korean Neurosurgical Society
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• 제56권5호
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• pp.375-382
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• 2014

Objective : To assess the effect of bone marrow mononuclear cells (BMMNCs) transplantation in the expression of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in spinal cord injury (SCI) in rats. Methods : BMMNCs were isolated from tibia and femur by a density gradient centrifugation. After establishment of acute transection SCI, rats were divided into experiment (BMMNCs), experiment control (0.1 M PBS infused) and sham surgery groups (laminectomy without any SCI). Locomotor function was assessed weekly for 5 weeks post-injury using BBB locomotor score and urinary bladder function daily for 4 weeks post-injury. Activity of NF-${\kappa}B$ in spinal cord was assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction. Results : At each time point post-injury, sham surgery group had significantly higher Basso, Beattie, Bresnahan locomotor and urinary bladder function scores than experiment and experiment control group (p<0.05). At subsequent time interval there were gradual improvement in both experiment and experiment control group, but experiment group had higher score in comparison to experiment control group (p<0.05). Comparisons were also made for expression of activated NF-${\kappa}B$ positive cells and level of NF-${\kappa}B$ messenger RNA in spinal cord at various time points between the groups. Activated NF-${\kappa}B$ immunoreactivity and level of NF-${\kappa}B$ mRNA expression were significantly higher in control group in comparison to experiment and sham surgery group (p<0.05). Conclusion : BMMNCs transplantation attenuates the expression of NF-${\kappa}B$ in injured spinal cord tissue and thus helps in recovery of neurological function in rat models with SCI.

• Asian Pacific Journal of Cancer Prevention
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• 제14권7호
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• pp.4441-4446
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• 2013

Cholangiocarcinoma (CCA) is a tumor of biliary ducts, which has a high mortality rate and dismal prognosis. Constitutively activation of the transcription factor nuclear factor kappa-B (NF-${\kappa}B$) has been previously demonstrated in CCA. It is therefore a potential target for CCA treatment. Effects of diethyldithiocarbamate (DDTC) on NF-${\kappa}B$-dependent apoptosis induction in cancer have been reported; however, anti-metastasis has never been addressed. Therefore, here the focus was on DDTC effects on CCA migration and adhesiond. Anti-proliferation, anti-migration and anti-adhesion activities were determined in CCA cell lines, along with p65 protein levels and function. NF-${\kappa}B$ target gene expression was determined by quantitative RT-PCR. DDTC inhibited CCA cell proliferation. Suppression of migration and adhesion were observed prior to anti-CCA proliferation. These effects were related to decreased p65, reduction in NF-${\kappa}B$ DNA binding, and impaired activity. Moreover, suppression of ICAM-1 expression supported NF-${\kappa}B$-dependent anti-metastatic effects of DDTC. Taken together, DDTC suppression of CCA migration and adhesion through inhibition of NF-${\kappa}B$ signaling pathway is suggested from the current study. This might be a promising treatment choice against CCA metastasis.

• Tuberculosis and Respiratory Diseases
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• 제48권5호
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• pp.682-698
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• 2000

Glucocorticoid receptor (GR) functions as a suppressor of inflammation by inhibiting the expression of many cytokine genes activated by NF-${\kappa}B$. The goal of this study is to investigate the mechanism by which GR repress NF-${\kappa}B$ activation in lung epithelial cells. We used A549 and BEAS-2B lung epithelia! cell lines. Using Ig$G{\kappa}$-NF-${\kappa}B$ luciferase reporter gene construct, we found that dexamethasone significantly suppressed TNF-$\alpha$-induced NF-${\kappa}B$ activation and the overexpression of GR showed dose-dependent reduction of TNF-$\alpha$-induced NF-${\kappa}B$ activity in both cell lines. However, DNA binding of NF-${\kappa}B$ induced by TNF-$\alpha$ in electromobility shift assay was not inhibited by dexamethasone. Super shift assay with anti-p65 antibody demonstrated the existence of p65 in NF-${\kappa}B$ complex induced by $\alpha$ Western blot showed that $I{\kappa}B{\alpha}$ degradation induced by TNF-$\alpha$ was not affected by dexamethasone and $I{\kappa}B{\kappa}$ was not induced by dexamethasone, neither. To evaluate p65 specific transactivation, we adopted co-transfection study of Gal4-p65TA1 or TA2 fusion protein expression system together with 5xGal4-luciferase vector. Co-transfection of GR with Gal4-p65TA1 or TA2 repressed luciferase activity profoundly to the level of 10-20% of p65TA1- or TA2-induced transcriptional activity. And this transrepressional effect was abolished by co-transfection of CBP of SRC-1 expression vectors. These results suggest that GR-mediated transrepression of NF-${\kappa}B$ in lung epithelial cells is through competing for binding to limiting amounts of transcriptional coactivators, CBP or SRC-1.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권2호
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• pp.94-100
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• 2006

Purpose: This study was done to confirm the role of NF-$\kappa$B in cisplatin-induced apoptosis of oral squamous cell carcinoma. Materials and Methods: Five cell lines originated from different oral cancer patients were tested for the apoptosis by the treatment of cisplatin. These cells showed different degree of cisplatin-resistance and the order is OSCC-2>OSCC-3>OSCC-5> OSCC-1>OSCC-4. OSCC-2 and OSCC-4 cells were assayed for the apoptosis by measuring DNA fragmentation and TUNEL staining after cisplatin treatment. While OSCC-4 cells showed apoptosis, OSCC-2 cells showed no or very slight apoptosis by cisplatin treatment. Next, It was determined whether NF-$\kappa$B activation is required in mediating cisplatin-induced apoptosis of OSCC-4. Result: The result was that elevated NF-$\kappa$B activity mediated cisplatin-induced apoptosis. Conclusion: In conclusion, these findings suggest that NF-$\kappa$B activation is essential to cisplatin-induced apoptosis and it may be involved in cisplatin resistance in OSCC cells.