Skip to main content
Log in

Withaferin A is a potent inhibitor of angiogenesis

  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

The medicinal plant Withania somnifera is widely researched for its anti-inflammatory, cardioactive and central nervous system effects. In Ayurveda, the major Traditional Indian medicine system, extracts from W. somnifera are distinctively employed for the treatment of arthritis and menstrual disorders. Because these conditions involve angiogenic processes we hypothesized that the W. somnifera extracts might contain angiogenesis inhibitors. We employed an endothelial cell-sprouting assay to monitor the purification of substances from W. somnifera root extracts and isolated as the active principle the previously known natural product withaferin A. We show that withaferin A inhibits human umbilical vein endothelial cell (HUVEC) sprouting in three-dimensional collagen-I matrix at doses which are relevant to NF-kappa B-inhibitory activity. Withaferin A inhibits cell proliferation in HUVECs (IC50=12 nM) at doses that are significantly lower than those required for tumor cell lines through a process associated with inhibition of cyclin D1 expression. We propose that the inhibition of NF-kappa B by withaferin A in HUVECs occurs by interference with the ubiquitin-mediated proteasome pathway as suggested by the increased levels of poly-ubiquitinated proteins. Finally, withaferin A is shown to exert potent anti-angiogenic activity in vivo at doses that are 500-fold lower than those previously reported to exert anti-tumor activity in vivo. In conclusion, our findings identify a novel mode of action of withaferin A, which highlights the potential use of this natural product for cancer treatment or prevention.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Folkman J, Merler E, Abernathy C, Williams G. Isolation of a tumor factor responsible or angiogenesis. J Exp Med 1971; 133: 275–88.

    Google Scholar 

  2. Colville-Nash PR, Scott DL. Angiogenesis and rheumatoid arthritis: Pathogenic and therapeutic implications. Ann Rheum Dis 1992; 517: 919–25.

    Google Scholar 

  3. Reynolds LP, Killilea SD, Redmer DA. Angiogenesis in the female reproductive system. FASEB J 1992; 6: 886–92.

    Google Scholar 

  4. Battegay EJ. Angiogenesis: Mechanistic insights, neovascular diseases, and therapeutic prospects. J Mol Med 1995; 73: 333–46.

    Google Scholar 

  5. Oliver SJ, Brahn E. Combination therapy in rheumatoid arthritis: The animal model perspective. J Rheumatol 1996; 44 (Suppl): 56–60.

    Google Scholar 

  6. Misra R. Modern drug development from traditional medicinal plants using radioligand receptor-binding assays. Med Res Rev 1998; 18: 383–402.

    Google Scholar 

  7. Mishra LC, Singh BB, Dagenais S. Scientific basis for the therapeutic use of Withania somnifera (Ashwagandha): Altern Med Rev 2000; 5: 334–46.

    Google Scholar 

  8. Upton R. Ashwagandha Root. Santa Cruz, California: American Herbal Pharmacopoeia 2000.

  9. Kulkarni RR, Patki PS, Jog VP et al. Treatment of osteoarthritis with a herbomineral formulation: a double-blind, placebo-controlled, crossover study. J Ethnopharmacol 1991; 33: 91–5.

    Google Scholar 

  10. Shohat B, Gitter S, Abraham A, Lavie D. Antitumor activity of withaferin A. Cancer Chemoth Rep 1967; 51: 271–6.

    Google Scholar 

  11. Korff T, Augustin HG. Integration of endothelial cells in multicellular spheroids prevents apoptosis and induces differentiation. J Cell Biol 1998; 143: 1341–52.

    Google Scholar 

  12. Burbridge MF, West DC. Rat Aortic Ring 3D Model of Angiogenesis in Vitro. In Clifford Murray J (ed): Methods in Molecular Medicine: Angiogenesis Protocols. Totowa, New Jersey: Humana Press 2001; 185.

    Google Scholar 

  13. Mohan R, Sivak J, Ashton P et al. Curcuminoids inhibit the angiogenic response stimulated by fibroblast growth factor-2, including expression of matrix metalloproteinase gelatinase B. J Biol Chem 2000; 275: 10405–12.

    Google Scholar 

  14. LaVallee TM, Zhan X, Herbstritt CJ et al. 2-Methoxyestradiol inhibits proliferation and induces apoptosis independently of estrogen receptors alpha and beta. Cancer Res 2002; 62: 3691–7.

    Google Scholar 

  15. Kiener PA, Marek F, Rodgers G et al. Induction of tumor necrosis factor, IFN-gamma, and acute lethality in mice by toxic and non-toxic forms of lipid A. J Immunol 1988; 141: 870–4.

    Google Scholar 

  16. Passanti A, Taylor RM, Pili R et al. A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab Invest 1992; 67: 519–28.

    Google Scholar 

  17. Klein S, de Fougerolles AR, Blaikie P et al. Alpha 5 beta 1 integrin activates an NF-kappa B-dependent program of gene expression important for angiogenesis and inflammation. Mol Cell Biol 2002; 16: 5912–22.

    Google Scholar 

  18. Shono T, Ono M, Izumi H et al. Involvement of the transcription factor NF-kappa B in tubular morphogenesis of human microvascular endothelial cells by oxidative stress. Mol Cell Biol 1996; 8: 4231–9.

    Google Scholar 

  19. Kupchan SM, Anderson WK, Bollinger P et al. Tumor inhibitors. XXXIX. Active principles of Acnistus arborescens. Isolation and structural and spectral studies of withaferin A and withacnistin. J Org Chem 1969; 34: 3858–66.

    Google Scholar 

  20. Pelletier SW, Mody NV, Nowacki J, Bhattacharyya J. Carbon-13 Nuclear magnetic resonance spectral analysis of naturally occurring withanolides and their derivatives. J Nat Prod 1979; 42: 512–21.

    Google Scholar 

  21. Glotter E. Withanolides and related ergostane-type steroids. Nat Prod Rep 1991; 8: 415–40.

    Google Scholar 

  22. Pollet I, Opina CJ, Zimmerman C et al. Bacterial lipopolysaccharide directly induces angiogenesis through TRAF6-mediated activation of NF-kappaB and c-Jun N-terminal kinase. Blood 2003; 102: 1740–2.

    Google Scholar 

  23. Palombella VJ, Rando OJ, Goldberg AL, Maniatis T. The ubiquitinproteasome pathway is required for processing the NF-kappa B1 precursor protein and the activation of NF-kappa B. Cell 1994; 78: 773–85.

    Google Scholar 

  24. Fuegner A. Inhibition of immunologically induced inflammation by the plant steroid withaferin A. Arzneimittel Forsch 1973; 23: 932–5.

    Google Scholar 

  25. Shohat B, Kirson I, Lavie D. Immunosuppressive activity of two plant steroidal lactones, withaferin A and withanolide E. Biomedicine 1978; 28: 18–24.

    Google Scholar 

  26. Furmanowa M, Gajdzis-Kuls D, Ruszkowska J et al. In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity. Planta Med 2001; 67: 146–9.

    Google Scholar 

  27. Sharada AC, Solomon FE, Devi PU et al. Antitumor and radiosensitizing effects of withaferin A on mouse Ehrlich ascites carcinoma in vivo. Acta Oncol 1996; 35: 95–100.

    Google Scholar 

  28. Brach MA, Henschler R, Mertelsmann RH, Herrmann F. Ionizing radiation induces expression and binding activity of the nuclear factor kappa B. J Clin Invest 1991; 88: 691–5.

    Google Scholar 

  29. Devi PU, Akagi K, Ostapenko V et al. Withaferin A: A new radiosensitizer from the Indian medicinal plant Withania somnifera. Int J Radiat Biol 1996; 69: 193–7.

    Google Scholar 

  30. Palombella VJ, Conner EM, Fuseler JW et al. Role of the proteasome and NF-kappa B in streptococcal cell wall-induced polyarthritis. Proc Natl Acad Sci USA 1998; 95: 15671–6.

    Google Scholar 

  31. Kwok BH, Koh B, Ndubuisi MI et al. The anti-inflammatory natural product parthenolide from the medicinal herb Feverfew directly binds to and inhibits IkappaB kinase. Chem Biol 2001; 8: 759–66.

    Google Scholar 

  32. Oikawa T, Sasaki T, Nakamura M et al. The proteasome is involved in angiogenesis. Biochem Biophys Res Commun 1998; 246: 243–8.

    Google Scholar 

  33. Mezquita J, Mezquita B, Pau M, Mezquita C. Down-regulation of Flt-1 gene expression by the proteasome inhibitor MG262. J Cell Biochem 2003; 89: 1138–47.

    Google Scholar 

  34. Kumeda SI, Deguchi A, Toi M et al. Induction of G1 arrest and selective growth inhibition by lactacystin in human umbilical vein endothelial cells. Anticancer Res 1999; 19: 3961–8.

    Google Scholar 

  35. Willems AR, Lanker S, Patton EE et al. Cdc53 targets phosphorylated G1 cyclins for degradation by the ubiquitin proteolytic pathway. Cell 1996; 86: 453–63.

    Google Scholar 

  36. Myung J, Kim KB, Crews CM. The ubiquitin-proteasome pathway and proteasome inhibitors. Med Res Rev 2001; 21: 245–73.

    Google Scholar 

  37. Sabichi AL, Demierre M-F, Hawk ET et al. Frontiers in Cancer Prevention Research. Cancer Res 2003; 63: 5649–55.

    Google Scholar 

  38. Tosetti F, Ferrari N, De Flora S, Albini A. 'Angioprevention': Angiogenesis is a common and key target for cancer chemopreventive agents. FASEB J 2002; 16: 2–14.

    Google Scholar 

  39. Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 2003; 10: 768–80.

    Google Scholar 

  40. Jayaprakasam B, Nair MG. Cyclooxygenase-2 enzyme inhibitory withanolides from Withania somnifera leaves. Tetrahedron 2003; 59: 841–9.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mohan, R., Hammers, H., Bargagna-mohan, P. et al. Withaferin A is a potent inhibitor of angiogenesis. Angiogenesis 7, 115–122 (2004). https://doi.org/10.1007/s10456-004-1026-3

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10456-004-1026-3

Navigation