Inflammation

, Volume 41, Issue 2, pp 722–731 | Cite as

ZEB2 Attenuates LPS-Induced Inflammation by the NF-κB Pathway in HK-2 Cells

  • Qi Ding
  • Yang Wang
  • Ai-ling Zhang
  • Tao Xu
  • Dan-dan Zhou
  • Xiao-Feng Li
  • Jun-Fa Yang
  • Lei Zhang
  • Xiao Wang
ORIGINAL ARTICLE
  • 164 Downloads

Abstract

As a transcription factor, zinc finger E-box binding homeobox 2 (ZEB2) includes multiple functional domains which interact with kinds of transcriptional co-effectors. It has been reported that ZEB2 was involved in signal transduction and multiple cellular functions. However, the functional role of ZEB2 in inflammation is still obscure. The aim of the current study is to explore the function of ZEB2 in inflammation cytokine secretion and the role of the nuclear factor-κB (NF-κB) signaling pathway in lipopolysaccharide (LPS)-induced human proximal tubule cell line (HK-2) cells. Our result demonstrated that expression of ZEB2 was significantly downregulated and expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was upregulated in response to LPS. Meanwhile, knockdown of ZEB2 by transfecting siRNA increased TNF-α and IL-6 secretion. Overexpression of ZEB2 resulted in a decrease of TNF-α and IL-6 secretion in HK-2 cells. Additionally, Western blot analysis indicated that ZEB2 suppressed the activation of the NF-κB signaling pathway via downregulating the levels of phosphorylated p65 and IκBα compared with LPS stimulation. Collectively, our data demonstrated that ZEB2 attenuated LPS-induced inflammation cytokine secretion possibly through suppressing the NF-κB signaling pathway.

KEY WORDS

ZEB2 TNF-α IL-6 NF-κB signaling pathway 

Abbreviations

AKI

Acute kidney injury

DMEM

Dulbecco’s modified Eagle’s medium

EMT

Epithelial-mesenchymal transition

HK-2 cell

Human kidney epithelial cell

IL-6

Interleukin-6

LPS

Lipopolysaccharide

NF-κB

Nuclear factor-κB

PCR

Polymerase chain reaction

p-p65

Phosphorylated p65

p-IκBα

Phosphorylated IκBα

PDTC

Pyrrolidine dithiocarbamate

siRNA

Small interfering RNA

TGF-β1

Transforming growth factor beta 1

TNF-α

Tumor necrosis factor-α

ZEB2

Zinc finger E-box-binding homeobox 2

Notes

Acknowledgements

This study was supported by the Chinese National Natural Science Foundation Project (81100302) and the Intercollegiate Key Projects of Nature Science of Anhui Province (KJ2017A169).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

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Fig. S1

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High resolution image (TIFF 435 kb)
10753_2017_727_MOESM2_ESM.pdf (122 kb)
ESM 2 (PDF 122 kb)

References

  1. 1.
    Hoste, E.A., S.M. Bagshaw, R. Bellomo, C.M. Cely, R. Colman, D.N. Cruz, et al. 2015. Epidemiology of acute kidney injury in critically ill patients: the multinational AKI-EPI study. Intensive Care Medicine 41: 1411–1423.CrossRefPubMedGoogle Scholar
  2. 2.
    Bellomo, R., J.A. Kellum, C. Ronco, R. Wald, J. Martensson, M. Maiden, et al. 2017. Acute kidney injury in sepsis. Intensive Care Medicine 43: 816–828.CrossRefPubMedGoogle Scholar
  3. 3.
    Shen, B., C. Zhao, C. Chen, Z. Li, Y. Li, Y. Tian, et al. 2017. Picroside II protects rat lung and A549 cell against LPS-induced inflammation by the NF-kappaB pathway. Inflammation 40: 752–761.CrossRefPubMedGoogle Scholar
  4. 4.
    Zarbock, A., H. Gomez, and J.A. Kellum. 2014. Sepsis-induced acute kidney injury revisited: pathophysiology, prevention and future therapies. Current Opinion in Critical Care 20: 588–595.CrossRefPubMedCentralPubMedGoogle Scholar
  5. 5.
    Rasouly, H.M., S. Kumar, S. Chan, A. Pisarek-Horowitz, R. Sharma, Q.J. Xi, et al. 2016. Loss of Zeb2 in mesenchyme-derived nephrons causes primary glomerulocystic disease. Kidney International 90: 1262–1273.CrossRefPubMedCentralPubMedGoogle Scholar
  6. 6.
    Hegarty, S.V., A.M. Sullivan, and G.W. O'Keeffe. 2015. Zeb2: a multifunctional regulator of nervous system development. Progress in Neurobiology 132: 81–95.CrossRefPubMedGoogle Scholar
  7. 7.
    Verschueren, K., J.E. Remacle, C. Collart, H. Kraft, B.S. Baker, P. Tylzanowski, et al. 1999. SIP1, a novel zinc finger/homeodomain repressor, interacts with Smad proteins and binds to 5′-CACCT sequences in candidate target genes. The Journal of Biological Chemistry 274: 20489–20498.CrossRefPubMedGoogle Scholar
  8. 8.
    Miyoshi, A., Y. Kitajima, K. Sumi, K. Sato, A. Hagiwara, Y. Koga, et al. 2004. Snail and SIP1 increase cancer invasion by upregulating MMP family in hepatocellular carcinoma cells. British Journal of Cancer 90: 1265–1273.CrossRefPubMedCentralPubMedGoogle Scholar
  9. 9.
    Elloul, S., M.B. Elstrand, J.M. Nesland, C.G. Trope, G. Kvalheim, I. Goldberg, et al. 2005. Snail, slug, and Smad-interacting protein 1 as novel parameters of disease aggressiveness in metastatic ovarian and breast carcinoma. Cancer 103: 1631–1643.CrossRefPubMedGoogle Scholar
  10. 10.
    Castro Alves, C., E. Rosivatz, C. Schott, R. Hollweck, I. Becker, M. Sarbia, et al. 2007. Slug is overexpressed in gastric carcinomas and may act synergistically with SIP1 and Snail in the down-regulation of E-cadherin. The Journal of Pathology 211: 507–515.CrossRefPubMedGoogle Scholar
  11. 11.
    Katoh, M., and M. Katoh. 2009. Integrative genomic analyses of ZEB2: transcriptional regulation of ZEB2 based on SMADs, ETS1, HIF1alpha, POU/OCT, and NF-kappaB. International Journal of Oncology 34: 1737–1742.CrossRefPubMedGoogle Scholar
  12. 12.
    Barbu, E.A., J. Zhang, E.H. Berenstein, J.R. Groves, L.M. Parks, and R.P. Siraganian. 2012. The transcription factor Zeb2 regulates signaling in mast cells. Journal of Immunology 188: 6278–6286.CrossRefGoogle Scholar
  13. 13.
    Oeckinghaus, A., and S. Ghosh. 2009. The NF-kappaB family of transcription factors and its regulation. Cold Spring Harbor Perspectives in Biology 1: a000034.CrossRefPubMedCentralPubMedGoogle Scholar
  14. 14.
    Wullaert, A., M.C. Bonnet, and M. Pasparakis. 2011. NF-kappaB in the regulation of epithelial homeostasis and inflammation. Cell Research 21: 146–158.CrossRefPubMedGoogle Scholar
  15. 15.
    Sun, S.C., J.H. Chang, and J. Jin. 2013. Regulation of nuclear factor-kappaB in autoimmunity. Trends in Immunology 34: 282–289.CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Wang Y. 2013. Attenuation of berberine on lipopolysaccharide-induced inflammatory and apoptosis responses in beta-cells via TLR4-independent JNK/NF-kappaB pathway. Pharmaceutical biology.Google Scholar
  17. 17.
    Wang, N., L. Mao, L. Yang, J. Zou, K. Liu, M. Liu, et al. 2017. Resveratrol protects against early polymicrobial sepsis-induced acute kidney injury through inhibiting endoplasmic reticulum stress-activated NF-kappaB pathway. Oncotarget 8: 36449–36461.PubMedCentralPubMedGoogle Scholar
  18. 18.
    Langenberg, C., S.M. Bagshaw, C.N. May, and R. Bellomo. 2008. The histopathology of septic acute kidney injury: a systematic review. Critical Care 12: R38.CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Gomez, H., and J.A. Kellum. 2016. Sepsis-induced acute kidney injury. Current Opinion in Critical Care 22: 546–553.CrossRefPubMedCentralPubMedGoogle Scholar
  20. 20.
    Hotchkiss, R.S., and I.E. Karl. 2003. The pathophysiology and treatment of sepsis. The New England Journal of Medicine 348: 138–150.CrossRefPubMedGoogle Scholar
  21. 21.
    Dai, Y., P. Jia, Y. Fang, H. Liu, X. Jiao, J.C. He, et al. 2016. miR-146a is essential for lipopolysaccharide (LPS)-induced cross-tolerance against kidney ischemia/reperfusion injury in mice. Scientific Reports 6: 27091.CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Zarjou, A., and A. Agarwal. 2011. Sepsis and acute kidney injury. Journal of the American Society of Nephrology : JASN 22: 999–1006.CrossRefPubMedGoogle Scholar
  23. 23.
    Souza, A.C., R.A. Volpini, M.H. Shimizu, T.R. Sanches, N.O. Camara, P. Semedo, et al. 2012. Erythropoietin prevents sepsis-related acute kidney injury in rats by inhibiting NF-kappaB and upregulating endothelial nitric oxide synthase. American Journal of Physiology Renal Physiology 302: F1045–F1054.CrossRefPubMedGoogle Scholar
  24. 24.
    Xiao, H., K. Tang, P. Liu, K. Chen, J. Hu, J. Zeng, et al. 2015. LncRNA MALAT1 functions as a competing endogenous RNA to regulate ZEB2 expression by sponging miR-200s in clear cell kidney carcinoma. Oncotarget 6: 38005–38015.PubMedCentralPubMedGoogle Scholar
  25. 25.
    Yu, C., P. Li, D. Qi, L. Wang, H.L. Qu, Y.J. Zhang, et al. 2017. Osthole protects sepsis-induced acute kidney injury via down-regulating NF-kappaB signal pathway. Oncotarget 8: 4796–4813.PubMedGoogle Scholar
  26. 26.
    Zhang, W., X. Lu, W. Wang, Z. Ding, Y. Fu, X. Zhou, et al. 2017. Inhibitory effects of emodin, thymol, and astragalin on Leptospira interrogans-induced inflammatory response in the uterine and endometrium epithelial cells of mice. Inflammation 40: 666–675.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Qi Ding
    • 1
    • 2
    • 3
  • Yang Wang
    • 1
    • 2
    • 3
  • Ai-ling Zhang
    • 1
    • 2
    • 3
  • Tao Xu
    • 1
    • 2
    • 3
  • Dan-dan Zhou
    • 1
    • 2
    • 3
  • Xiao-Feng Li
    • 1
    • 2
    • 3
  • Jun-Fa Yang
    • 1
    • 2
    • 3
  • Lei Zhang
    • 1
    • 2
    • 3
  • Xiao Wang
    • 4
  1. 1.School of PharmacyAnhui Medical UniversityHefeiChina
  2. 2.Key Laboratory of Major Autoimmune Disease, Anhui Province; School of PharmacyAnhui Medical UniversityHefeiChina
  3. 3.The Key Laboratory of Anti-inflammatory and Immune Medicines, Ministry of EducationAnhui Medical UniversityHefeiChina
  4. 4.Department of RadiologyThe First Affiliated Hospital of Anhui Medical UniversityHefeiChina

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