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Cucurbitacin E exhibits anti-inflammatory effect in RAW 264.7 cells via suppression of NF-κB nuclear translocation

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Abstract

Objective

Cucurbitacin E (CuE), a triterpenoid compound isolated from Cucurbitaceae plants, possesses a wide range of biological activities including anti-inflammatory properties. The present study aimed to investigate the anti-inflammatory effect of CuE and the underlying mechanism of action.

Methods

The anti-inflammatory effect of CuE was evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Cell proliferation was assessed using a modified MTT assay. Cell cycle distribution was analyzed by propidium iodide staining. The actin cytoskeleton was examined by immunofluorescent staining. The expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β was determined by intracellular cytokine staining. G-actin level and nuclear factor (NF)-κB nuclear translocation were detected by immunoblotting.

Results

CuE inhibited cell proliferation and induced cell cycle arrest at G2/M phase in RAW 264.7 cells. CuE also suppressed LPS-induced cell spreading and pseudopodia formation. These effects were associated with decreased G-actin level and severe actin aggregation. Moreover, CuE significantly inhibited both TNF-α and IL-1β production in LPS-stimulated RAW 264.7 cells. This was likely mediated by suppressing LPS-induced nuclear translocation of NF-κB, a critical transcription factor responsible for pro-inflammatory cytokine expression.

Conclusion

CuE displayed anti-inflammatory effects through suppression of NF-κB nuclear translocation leading to a decreased expression of TNF-α and IL-1β in LPS-stimulated RAW 264.7 cells.

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References

  1. Chan KT, Li K, Liu SL, Chu KH, Toh M, Xie WD. Cucurbitacin B inhibits STAT3 and the Raf/MEK/ERK pathway in leukemia cell line K562. Cancer Lett. 2010;289:46–52.

    Article  PubMed  CAS  Google Scholar 

  2. Dong Y, Lu B, Zhang X, Zhang J, Lai L, Li D, Wu Y, Song Y, Luo J, Pang X, Yi Z, Liu M. Cucurbitacin E, a tetracyclic triterpenes compound from Chinese medicine, inhibits tumor angiogenesis through VEGFR2-mediated Jak2-STAT3 signaling pathway. Carcinogenesis. 2010;31:2097–104.

    Article  PubMed  CAS  Google Scholar 

  3. Peters RR, Farias MR, Ribeiro-do-Valle RM. Anti-inflammatory and analgesic effects of cucurbitacins from Wilbrandia ebracteata. Planta Med. 1997;63:525–8.

    Article  PubMed  CAS  Google Scholar 

  4. Duncan KL, Duncan MD, Alley MC, Sausville EA. Cucurbitacin E-induced disruption of the actin and vimentin cytoskeleton in prostate carcinoma cells. Biochem Pharmacol. 1996;52:1553–60.

    Article  PubMed  CAS  Google Scholar 

  5. Abdelwahab SI, Hassan LE, Sirat HM, Yagi SM, Koko WS, Mohan S, Taha MM, Ahmad S, Chuen CS, Narrima P, Rais MM, Hadi AH. Anti-inflammatory activities of cucurbitacin E isolated from Citrullus lanatus var. citroides: role of reactive nitrogen species and cyclooxygenase enzyme inhibition. Fitoterapia. 2011;82:1190–7.

    Article  PubMed  CAS  Google Scholar 

  6. Salminen A, Huuskonen J, Ojala J, Kauppinen A, Kaarniranta K, Suuronen T. Activation of innate immunity system during aging: NF-kB signaling is the molecular culprit of inflamm-aging. Ageing Res Rev. 2008;7:83–105.

    Article  PubMed  CAS  Google Scholar 

  7. Sen R, Baltimore D. Inducibility of K immunoglobulin enhancer-binding protein NF-κB by a posttranslational mechanism. Cell. 1986;47:921–8.

    Article  PubMed  CAS  Google Scholar 

  8. Karin M, Staudt L. NF-κB a network hub controlling immunity, inflammation, and cancer: a subject collection from Cold Spring Harbor perspectives in biology. Cold Spring Harbor, Cold Spring Harbor Laboratory Press, NY; 2010. viii, p 271.

  9. Karin M, Greten FR. NF kappa B: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol. 2005;5:749–59.

    Article  PubMed  CAS  Google Scholar 

  10. Liou H-C. NF-κB/Rel transcription factor family. Georgetown, Tex. New York: Landes Bioscience/Eurekah.com, Springer Science+Business Media, 2006. p 185.

  11. Jin HR, Jin X, Dat NT, Lee JJ. Cucurbitacin B suppresses the transactivation activity of RelA/p65. J Cell Biochem. 2011;112:1643–50.

    Article  PubMed  CAS  Google Scholar 

  12. Zhang Y, Ouyang D, Xu L, Ji Y, Zha Q, Cai J, He X. Cucurbitacin B induces rapid depletion of the G-actin pool through reactive oxygen species-dependent actin aggregation in melanoma cells. Acta Biochim Biophys Sin. 2011;43:556–67.

    Article  PubMed  CAS  Google Scholar 

  13. Zhang YT, Ouyang DY, Xu LH, Ji YH, Zha QB, Cai JY, He XH. Cucurbitacin B induces rapid depletion of the G-actin pool through reactive oxygen species-dependent actin aggregation in melanoma cells. Acta Biochim Biophys Sin. 2011;43:556–67.

    Article  PubMed  CAS  Google Scholar 

  14. Ouyang D, Zhang Y, Xu L, Li J, Zha Q, He X. Histone deacetylase inhibitor valproic acid sensitizes B16F10 melanoma cells to cucurbitacin B treatment. Acta Biochim Biophys Sin. 2011;43:487–95.

    Article  PubMed  CAS  Google Scholar 

  15. Duncan MD, Duncan KL. Cucurbitacin E targets proliferating endothelia. J Surg Res. 1997;69:55–60.

    Article  PubMed  CAS  Google Scholar 

  16. Jayaprakasam B, Seeram NP, Nair MG. Anticancer and antiinflammatory activities of cucurbitacins from Cucurbita andreana. Cancer Lett. 2003;189:11–6.

    Article  PubMed  CAS  Google Scholar 

  17. Ziegler-Heitbrock H, Sternsdorf T, Liese J, Belohradsky B, Weber C, Wedel A, Schreck R, Bauerle P, Strobel M. Pyrrolidine dithiocarbamate inhibits NF-κB mobilization and TNF production in human monocytes. J Immunol. 1993;151:6986–93.

    PubMed  CAS  Google Scholar 

  18. Varfolomeev EE, Ashkenazi A. Tumor necrosis factor: an apoptosis JuNKie? Cell. 2004;116:491–7.

    Article  PubMed  CAS  Google Scholar 

  19. Nakashima S, Matsuda H, Kurume A, Oda Y, Nakamura S, Yamashita M, Yoshikawa M. Cucurbitacin E as a new inhibitor of cofilin phosphorylation in human leukemia U937 cells. Bioorg Med Chem Lett. 2010;20:2994–7.

    Article  PubMed  CAS  Google Scholar 

  20. Sorensen PM, Iacob RE, Fritzsche M, Engen JR, Brieher WM, Charras G. Eggert US. ACS Chem Biol: The natural product cucurbitacin E inhibits depolymerization of actin filaments; 2012.

    Google Scholar 

  21. Momma K, Masuzawa Y, Nakai N, Chujo M, Murakami A, Kioka N, Kiyama Y, Akita T, Nagao M. Direct interaction of Cucurbitacin E isolated from Alsomitra macrocarpa to actin filament. Cytotechnology. 2008;56:33–9.

    Article  PubMed  Google Scholar 

  22. Huang WW, Yang JS, Lin MW, Chen PY, Chiou SM, Chueh FS, Lan YH, Pai SJ, Tsuzuki M, Ho WJ, Chung JG. Cucurbitacin E induces G(2)/M phase arrest through STAT3/p53/p21 signaling and provokes apoptosis via Fas/CD95 and mitochondria-dependent pathways in human bladder cancer T24 cells. Evid Based Complement Alternat Med. 2012;2012:952762.

    PubMed  Google Scholar 

  23. Li Y, Wang R, Ma E, Deng Y, Wang X, Xiao J, Jing Y. The induction of G2/M cell-cycle arrest and apoptosis by cucurbitacin E is associated with increased phosphorylation of eIF2α in leukemia cells. Anticancer Drugs. 2010;21:389–400.

    Article  PubMed  CAS  Google Scholar 

  24. Beutler B, Cerami A. The biology of cachectin/TNF–a primary mediator of the host response. Ann Rev Immunol. 1989;7:625–55.

    Article  CAS  Google Scholar 

  25. Verstrepen L, Bekaert T, Chau TL, Tavernier J, Chariot A, Beyaert R. TLR-4, IL-1R and TNF-R signaling to NF-κB: variations on a common theme. Cell Mol Life Sci. 2008;65:2964–78.

    Article  PubMed  CAS  Google Scholar 

  26. Yang SK, Wang YC, Chao CC, Chuang YJ, Lan CY, Chen BS. Dynamic cross-talk analysis among TNF-R, TLR-4 and IL-1R signalings in TNF-α-induced inflammatory responses. BMC Med Genomics. 2010;3:19.

    Article  PubMed  Google Scholar 

  27. Lee KC, Chang HH, Chung YH, Lee TY. Andrographolide acts as an anti-inflammatory agent in LPS-stimulated RAW264.7 macrophages by inhibiting STAT3-mediated suppression of the NF-kappaB pathway. J Ethnopharmacol. 2011;135:678–84.

    Article  PubMed  CAS  Google Scholar 

  28. O’Neill LAJ, Kaltschmidt C. NF-kB: a crucial transcription factor for glial and neuronal cell function. Trends Neurosci. 1997;20:252–8.

    Article  PubMed  Google Scholar 

  29. Shrum CK, Defrancisco D, Meffert MK. Stimulated nuclear translocation of NF-κB and shuttling differentially depend on dynein and the dynactin complex. Proc Natl Acad Sci USA. 2009;106:2647–52.

    Article  PubMed  CAS  Google Scholar 

  30. Rosette C, Karin M. Cytoskeletal control of gene expression: depolymerization of microtubules activates NF-kappa B. J Cell Biol. 1995;128:1111–9.

    Article  PubMed  CAS  Google Scholar 

  31. Banan A, Keshavarzian A, Zhang L, Shaikh M, Forsyth C, Tang Y, Fields J. NF-κB activation as a key mechanism in ethanol-induced disruption of the F-actin cytoskeleton and monolayer barrier integrity in intestinal epithelium. Alcohol. 2007;41:447–60.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work is supported by grants from the National Natural Science Foundation of China (No.81173604), the Specialized Research Program of “Twelfth Five-Year Plan” of China (No.2011ZX09307-303-03), and the Fundamental Research Funds for the Central Universities (No.21612411).

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Authors and Affiliations

  1. Department of Immunobiology, Institute of Tissue Transplantation and Immunology, Jinan University, 601 Huangpu Dadao West, Guangzhou, 510630, China

    Jing Qiao, Jian He, Dong-yun Ouyang & Xian-hui He

  2. Department of Cell Biology, Jinan University, Guangzhou, 510630, China

    Li-hui Xu

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  1. Jing Qiao
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  2. Li-hui Xu
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  3. Jian He
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Correspondence to Xian-hui He.

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Responsible Editor: Liwu Li.

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Qiao, J., Xu, Lh., He, J. et al. Cucurbitacin E exhibits anti-inflammatory effect in RAW 264.7 cells via suppression of NF-κB nuclear translocation. Inflamm. Res. 62, 461–469 (2013). https://doi.org/10.1007/s00011-013-0598-z

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  • DOI: https://doi.org/10.1007/s00011-013-0598-z

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