Lipoteichoic acid upregulates NF-κB and proinflammatory cytokines by modulating β-catenin in bronchial epithelial cells

JANG, JAEWOONG,KIM, WONYONG,KIM, KIJEONG,CHUNG, SANG-IN,SHIM, YAE JIE,KIM, SEOK-MIN,YOON, YOOSIK Spandidos Publications 2015 MOLECULAR MEDICINE REPORTS Vol.12 No.3

<P>Lipoteichoic acid (LTA) is a major cell wall component and virulence factor of gram-positive bacteria. The present study investigated the LTA?induced inflammatory response of BEAS?2B human bronchial epithelial cells, and detected the expression levels of proinflammatory cytokines interleukin (IL)?6, IL?8, IL?1β, tumour necrosis factor?α and monocyte chemotactic protein?1, the upregulation of NF?κB, and the phosphorylation and degradation of I?κB. During the LTA?induced inflammatory response of the BEAS?2B human bronchial epithelial cells, the activity levels of the β?catenin?dependent promoter, and the protein expression levels of β?catenin were significantly upregulated, whereas β?catenin phosphorylation and the expression levels of AXIN were significantly downregulated. Following knockdown of β?catenin by small interfering (si)RNA transfection, both the LTA-induced protein expression levels of NF?κB and the LTA-induced activity levels of the NF?κB?dependent promoter were significantly reduced. Similarly, a marked reduction in I?κB phosphorylation and degradation was observed following β?catenin knockdown. The expression levels of the LTA?induced proinflammatory cytokines were also significantly reduced following β?catenin siRNA. These results suggest that β?catenin has a significant role in the regulation of NF?κB activity and proinflammatory cytokine expression during the LTA-induced inflammatory response of bronchial epithelial cells.</P>

A calpain inhibitor protects against fractalkine production in lipopolysaccharide-treated endothelial cells

( Jaewoong Jang ),( Yoosik Yoon ),( Dong-jin Oh ) 대한신장학회 2017 Kidney Research and Clinical Practice Vol.36 No.3

Background: Fractalkine (CX3CL1) is a chemokine with a unique CX3C motif and is produced by endothelial cells stimulated with lipopolysaccharide (LPS), tumor necrosis factor (TNF)-α, interleukin (IL)-1, and interferon-γ. There have been several reports that the caspase/calpain system is activated in endotoxemia, which leads to cellular apoptosis and acute inflammatory processes. We aimed to determine the role of the caspase/calpain system in cell viability and regulation of fractalkine production in LPS-treated endothelial cells. Methods: Human umbilical vein endothelial cells (HUVECs) were stimulated with 0.01-100 μg/mL of LPS to determine cell viability. The changes of CX3CL1 expression were compared in control, LPS (1 μg/mL)-, IL-1α (1 μg/ mL)-, and IL-1β (1 μg/mL)-treated HUVECs. Cell viability and CX3CL1 production were compared with 50 μM of inhibitors of caspase-1, caspase-3, caspase-9, and calpain in LPS-treated HUVECs. Results: Cell viability was significantly decreased from 1 to 100 μg/mL of LPS. Cell viability was significantly restored with inhibitors of caspase-1, caspase-3, caspase-9, and calpain in LPS-treated HUVECs. The expression of CX3CL1 was highest in IL-1β-treated HUVECs. CX3CL1 production was highly inhibited with a calpain inhibitor and significantly decreased with the individual inhibitors of caspase-1, caspase-3, and caspase-9. Conclusion: The caspase/calpain system is an important modulator of cell viability and CX3CL1 production in LPStreated endothelial cells.

β-catenin mediates the inflammatory cytokine expression induced by the Der p 1 house dust mite allergen

JANG, JAEWOONG,HA, JONG-HYEOK,KIM, SEOK-MIN,KIM, WONYONG,KIM, KIJEONG,CHUNG, SANG-IN,YOON, YOOSIK SPANDIDOS PUBLICATIONS 2014 MOLECULAR MEDICINE REPORTS Vol.9 No.2

β-catenin regulates NF-κB activity and inflammatory cytokine expression in bronchial epithelial cells treated with lipopolysaccharide.

Jang, Jaewoong,Ha, Jong-Hyeok,Chung, Sang-In,Yoon, Yoosik D.A. Spandidos 2014 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.34 No.2

<P>In the present study, we demonstrate that lipopolysaccharide (LPS) induces the expression of inflammatory cytokines, including interleukin (IL)-6, IL-8, IL-1β, tumor necrosis factor (TNF)-α and monocyte chemoattractant protein (MCP)-1 in BEAS-2B human bronchial epithelial cells in a dose- and time-dependent manner. This increase was accompanied by an increased activity of nuclear factor (NF)?κB. When the expression of β-catenin was analyzed following treatment with LPS, the mRNA level was unaltered; however, the β-catenin protein levels increased with a decrease in phosphorylation at the serine 33/37 residues. Nuclear β-catenin protein levels also increased along with the reporter activity of a β-catenin-responsive TOPFlash vector. To elucidate the regulatory role of β-catenin in the LPS-induced inflammatory response of bronchial epithelial cells, β-catenin production was knocked down using siRNA. Our results revealed that β-catenin protein levels and TOPFlash vector reporter activity were reduced to basal levels by siRNA transfection. In this experimental condition, NF-κB activity, measured by enzyme-linked immunosorbent assay (ELISA), electrophoretic mobility shift assay (EMSA) and an NF-κB responsive reporter assay, was reduced to basal levels. Similarly, LPS-induced inflammatory cytokine expression was reduced almost to basal levels following transfection with β-catenin siRNA. These results demonstrate that β-catenin positively regulates NF-κB activity, as well as the expression of inflammatory cytokines in the inflammatory response of LPS-treated bronchial epithelial cells.</P>

Wnt-C59 inhibits proinflammatory cytokine expression by reducing the interaction between -catenin and NF-κB in LPS-stimulated epithelial and macrophage cells

Jaewoong Jang,Jaewon Song,Inae Sim,Yoosik Yoon 대한생리학회-대한약리학회 2021 The Korean Journal of Physiology & Pharmacology Vol.25 No.4

Dysregulation of the Wnt pathway causes various diseases including cancer, Parkinson’s disease, Alzheimer’s disease, schizophrenia, osteoporosis, obesity and chronic kidney diseases. The modulation of dysregulated Wnt pathway is abso-lutely necessary. In the present study, we evaluated the anti-inflammatory effect and the mechanism of action of Wnt-C59, a Wnt signaling inhibitor, in lipopolysaccharide (LPS)-stimulated epithelial cells and macrophage cells. Wnt-C59 showed a dose-de-pendent anti-inflammatory effect by suppressing the expression of proinflammatory cytokines including IL6, CCL2, IL1A, IL1B, and TNF in LPS-stimulated cells. The dysreg-ulation of the Wnt/β-catenin pathway in LPS stimulated cells was suppressed by Wnt-C59 treatment. The level of β-catenin, the executor protein of Wnt/β-catenin path-way, was elevated by LPS and suppressed by Wnt-C59. Overexpression of β-catenin rescued the suppressive effect of Wnt-C59 on proinflammatory cytokine expression and nuclear factor-kappa B (NF-κB) activity. We found that the interaction between β-catenin and NF-κB, measured by co-immunoprecipitation assay, was elevated by LPS and suppressed by Wnt-C59 treatment. Both NF-κB activity for its target DNA binding and the reporter activity of NF-κB-responsive promoter showed identical patterns with the interaction between β-catenin and NF-κB. Altogether, our findings suggest that the anti-inflammatory effect of Wnt-C59 is mediated by the reduction of the cellular level of β-catenin and the interaction between β-catenin and NF-κB, which results in the suppressions of the NF-κB activity and proinflammatory cytokine expression.

LGK974 suppresses lipopolysaccharide-induced endotoxemia in mice by modulating the crosstalk between the Wnt/β-catenin and NF-κB pathways

Jang Jaewoong,Song Jaewon,Lee Hyunji,Sim Inae,Kwon Young V.,Jho Eek-hoon,Yoon Yoosik 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Endotoxemia, a type of sepsis caused by gram-negative bacterial endotoxin [i.e., lipopolysaccharide (LPS)], is associated with manifestations such as cytokine storm; failure of multiple organs, including the liver; and a high mortality rate. We investigated the effect and mechanism of action of LGK974, a Wnt signaling inhibitor, in mice with LPS-induced endotoxemia, an animal model of sepsis. LGK974 significantly and dose-dependently increased the survival rate and reduced plasma cytokine levels in mice with LPS-induced endotoxemia. Transcriptome analysis of liver tissues revealed significant changes in the expression of genes associated with the Wnt pathway as well as cytokine and NF-κB signaling during endotoxemia. LGK974 treatment suppressed the activation of NF-κB signaling and cytokine expression as well as the Wnt/β-catenin pathway in the livers of endotoxemic mice. Coimmunoprecipitation of phospho-IκB and β-transducin repeat-containing protein (β-TrCP) was increased in the livers of endotoxemic mice but was reduced by LGK974 treatment. Moreover, LGK974 treatment decreased the coimmunoprecipitation and colocalization of β-catenin and NF-κB, which were elevated in the livers of endotoxemic mice. Our results reveal crosstalk between the Wnt/β-catenin and NF-κB pathways via interactions between β-TrCP and phospho-IκB and between β-catenin and NF-κB during endotoxemia. The results of this study strongly suggest that the crosstalk between the Wnt/β-catenin and NF-κB pathways contributes to the mutual activation of these two pathways during endotoxemia, which results in amplified cytokine production, liver damage and death, and that LGK974 suppresses this vicious amplification cycle by reducing the crosstalk between these two pathways.

Berberine activates AMPK to suppress proteolytic processing, nuclear translocation and target DNA binding of SREBP-1c in 3T3-L1 adipocytes

Jang, Jaewoong,Jung, Yoonju,Seo, Seong Jun,Kim, Seok-Min,Shim, Yae Jie,Cho, Soo Hyun,Chung, Sang-In,Yoon, Yoosik SPANDIDOS PUBLICATIONS 2017 MOLECULAR MEDICINE REPORTS Vol.15 No.6

<P>AMP-activated protein kinase (AMPK) and sterol regulatory element binding protein (SREBP)-1c are major therapeutic targets in the treatment of metabolic diseases. In the present study, the fat-reducing mechanisms of berberine (BBR), a natural isoquinoline, was investigated by examining the AMPK-mediated modulation of SREBP-1c in 3T3-L1 adipocytes. BBR activated AMPK in a dose- and time-dependent manner, and increased the phosphorylation of the 125-kDa precursor form of SREBP-1c, which suppressed its proteolytic processing into the mature 68-kDa form and its subsequent nuclear translocation. The binding of nuclear SREBP-1c to its E-box motif-containing target DNA sequence was decreased following treatment with BBR, which led to a decrease in the expression of lipogenic genes and subsequently reduced intracellular fat accumulation. Transfection with AMPKα1 siRNA, and not control siRNA, inhibited BBR-induced phosphorylation of the 125-kDa SREBP-1c, which confirmed that AMPK was responsible for phosphorylating SREBP-1c. AMPKα1 siRNA transfection rescued the proteolytic processing, nuclear translocation and target DNA binding of SREBP-1c that had been suppressed by BBR. In addition, BBR-induced suppression of lipogenic gene expression and intracellular fat accumulation were rescued by AMPKα1 siRNA transfection. In conclusion, the results of the present study demonstrate that BBR activates AMPK to induce phosphorylation of SREBP-1c, thereby suppressing proteolytic processing, nuclear translocation and target DNA binding of SREBP-1c, which leads to a reduction in lipogenic gene expression and intracellular fat accumulation. The results of the present study indicate that BBR may be a potential candidate for the development of drugs to treat obesity.</P>

동적 수밀부 표면처리에 대한 실험적 연구

장재웅(Jaewoong Jang),김재기(Jaeki Kim),신명섭(Myungsub Shin) 대한기계학회 2017 대한기계학회 춘추학술대회 Vol.2017 No.11

There are many treatments on surface to improve hardness. One of them. tungsten-carbide coating by H.V.O.F has good hardness and wear resistance. But after rotating shaft(Al) coated tungsten-carbide to seal rotating surface operates in sea water, corrosion occurs on sealing surface coated WC H.V.O.F. Considering corrosion, hardness, roughness of surface and porductivity, hard anodizing apply on sealing surface by verification test and research.

A sketch-based interface to modify and reproduce motion sequences

Jang, Hyunho,Jeon, Jaewoong,Sohn, Eisung,Lim, Soon-Bum,Choy, Yoon-Chul Springer-Verlag 2014 MULTIMEDIA TOOLS AND APPLICATIONS Vol.72 No.1

Essential role of flotillin-1 palmitoylation in the intracellular localization and signaling function of IGF-1 receptor.

Jang, Donghwan,Kwon, Hayeong,Jeong, Kyuho,Lee, Jaewoong,Pak, Yunbae The Company of Biologists Ltd. 2015 Journal of cell science Vol.128 No.11

<P>Here, we explored flotillin-1-mediated regulation of insulin-like growth factor-1 (IGF-1) signaling. Flotillin-1-deficient cells exhibited a reduction in the activation of IGF-1 receptor (IGF-1R), ERK1/2 and Akt pathways, and the transcriptional activation of Elk-1 and the proliferation in response to IGF-1 were reduced in these cells. We found that IGF-1-independent flotillin-1 palmitoylation at Cys34 in the endoplasmic reticulum (ER) was required for the ER exit and the plasma membrane localization of flotillin-1 and IGF-1R. IGF-1-dependent depalmitoylation and repalmitoylation of flotillin-1 sustained tyrosine kinase activation of the plasma-membrane-targeted IGF-1R. Dysfunction and blocking the turnover of flotillin-1 palmitoylation abrogated cancer cell proliferation after IGF-1R signaling activation. Our data show that flotillin-1 palmitoylation is a new mechanism by which the intracellular localization and activation of IGF-1R are controlled.</P>