Protein & Cell

, Volume 3, Issue 7, pp 535–544 | Cite as

Curcumin induces differentiation of embryonic stem cells through possible modulation of nitric oxide-cyclic GMP pathway

  • Kalpana MujooEmail author
  • Lubov E. Nikonoff
  • Vladislav G Sharin
  • Nathan S. Bryan
  • Alexander Y. Kots
  • Ferid Murad
Research Article


Curcumin, an active ingredient of dietary spice used in curry, has been shown to exhibit anti-oxidant, anti-inflammatory and anti-proliferative properties. Using EB directed differentiation protocol of H-9 human embryonic stem (ES) cells; we evaluated the effect of curcumin (0–20 μmol/L) in enhancing such differentiation. Our results using real time PCR, western blotting and immunostaining demonstrated that curcumin significantly increased the gene expression and protein levels of cardiac specific transcription factor NKx2.5, cardiac troponin I, myosin heavy chain, and endothelial nitric oxide synthase during ES cell differentiation. Furthermore, an NO donor enhanced the curcumin-mediated induction of NKx2.5 and other cardiac specific proteins. Incubation of cells with curcumin led to a dose dependent increase in intracellular nitrite to the same extent as giving an authentic NO donor. Functional assay for second messenger(s) cyclic AMP (cAMP) and cyclic GMP (cGMP) revealed that continuous presence of curcumin in differentiated cells induced a decrease in the baseline levels of cAMP but it significantly elevated baseline contents of cGMP. Curcumin addition to a cell free assay significantly suppressed cAMP and cGMP degradation in the extracts while long term treatment of intact cells with curcumin increased the rates of cAMP and cGMP degradation suggesting that this might be due to direct suppression of some cyclic nucleotide-degrading enzyme (phosphodiesterase) by curcumin. These studies demonstrate that polyphenol curcumin may be involved in differentiation of ES cells partly due to manipulation of nitric oxide signaling.


curcumin nitric oxide cyclic GMP embryonic stem cells 


  1. Aggarwal, B. B., and Sung, S. (2008). Pharmacological basis for the role of curcumin in chronic diseases: an age old spice with modern targets. Trend Pharmacol Sci 30, 85–94.CrossRefGoogle Scholar
  2. Aggarwal, B.B., Sundaram, C., Malani, N., and Ichikawa, H. (2007) Curcumin: the Indian solid gold. Adv Exp Med Biol 595, 1–75.CrossRefGoogle Scholar
  3. Bryan, N.S. and Grisham, M.B. (2007). Methods to detect nitric oxide and its metabolites in biological samples. Free Rad Biol Med 43, 645–657.CrossRefGoogle Scholar
  4. Fang, X-D., Yang, F. L., Zhu, L., Shen, Y-L, Chen, Y-Y. (2009). Curcumin ameliorates high glucose-induced acute vascular endothelial dysfunction in rat thoracic aorta. Clin Expl Pharmacol Physiol 36, 1177–1182.CrossRefGoogle Scholar
  5. Goldstein, B., Rogelj, S., Siegel, S., Farmer, S.R., and Niles, R.M. (1990). Cyclic adenosine monophosphate-mediated induction of F9 teratocarcinoma differentiation in absence of retinoic acid. J Cell Physiol 143, 205–212.CrossRefGoogle Scholar
  6. Kang S-K., Cha, S-H., and Jeon, H-G. (2006). Curcumin-induced histone hypoacetylation enhances caspase-3-dependent glioma cell death and neurogenesis of neural progenitor cells. Stem Cells Develop 15, 165–174.CrossRefGoogle Scholar
  7. Kim, S. J., Son, T.G., Park, H.R., Park, M., Kim, M-S., Kim, H-S., Chung, H.Y., Mattson, M.P., Lee, J. (2008). Curcumin stimulates proliferation of embryonic neural progenitor cells and neurogenesis in the adult hippocampus. J Biol Chem 283, 14497–14505.CrossRefGoogle Scholar
  8. Kots, A. Y., Choi, B-K., Estrella-Jimenez, M.E., Warren, C.A., Gilbertson, S.R., Guerrant, R.L., and Murad, F. (2008). Pyridopyrimidine derivatives as inhibitors of cyclic nucleotide synthesis: application for treatment of diarrhea. Procs Nat Acad Sci U S A 105, 8440–8445.CrossRefGoogle Scholar
  9. Krumenacker, J.S., Katsuki, S., Kots, A.Y., and Murad, F. (2006). Differential expression of genes involved in cGMP-dependent nitric oxide signaling in murine embryonic stem (ES) cells and cell-derived cardiomyocytes. Nitric Oxide 14, 1–11.CrossRefGoogle Scholar
  10. Lin, T., Chao, C., Saito, S., Mazur, S.J., Murphy, M.E., Appella, E. and Xu, Y. (2005). p53 induces differentiation of mouse embryonic stem cells by suppressing nanog expression. Nature Cell Biol 7, 165–171.CrossRefGoogle Scholar
  11. Livak, K.J. and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2 -ΔΔC T method. Methods 25, 25402–25408CrossRefGoogle Scholar
  12. Lodha, R. and Bagga, A. (2000). Traditional Indian systems of Medicine. Ann Acad Med Singapore 29, 37–41.Google Scholar
  13. Morimoto, T., Sunagawa, Y., Kawamura, T., Takaya, T., Wada, H., Nagasawa, A., Komeda, M., Fujita, M. Shimatsu, A., Kita, T., Hasegawa, K. (2008). The dietary compound curcumin inhibits p300 histone acetyl transferase activity and prevents heart failure in rats. J Clin Invest 118, 868–878.Google Scholar
  14. Mujoo, K., Krumenacker, J.S., Wada, Y. and Murad, F. (2006). Differential expression of nitric oxide signaling components in undifferentiated and differentiated human embryonic stem cells. Stem Cells Develop 15, 779–787.CrossRefGoogle Scholar
  15. Mujoo, K., Sharin, V.G., Bryan, N.S., Krumenacker, J.S., Sloan, C., Parveen, S., Kots, A.Y. and Murad, F. (2008). Role of nitric oxide signaling components in differentiation of embryonic stem cells into myocardial cells. Proc Natl Acad Sci U S A 105, 18924–18929.CrossRefGoogle Scholar
  16. Mujoo K, Krumenacker JS, Murad F. (2011). Nitric oxide-cyclic GMP signaling in stem cell differentiation. Free Rad Bio Med 51, 2150–2157CrossRefGoogle Scholar
  17. Murad, F. (2006). Shattuck Lecture. Nitric oxide and cyclic GMP in cell signaling and drug development. New Eng J Med 355, 2003–2011.CrossRefGoogle Scholar
  18. Pan, W., Quarles, L.D, Song, L.H., Yu, Y.H., Jiao, C., Tang, H.B., Jiang, C.H., Deng, H.W., Li, Y.J., Zhou, H.H., Xiao, Z.S. (2005). Genistein stimulates the osteoblastic differentiation via NO/cGMP in bone marrow culture. J Cell Biochem 94, 307–316.CrossRefGoogle Scholar
  19. Sandur, S.K., Ichikawa, H., Pandey, M.K., Kunnumakkara, A.B., Sung, S., Sethi, G. and Aggarwal, B.B. (2007). Role of pro-oxidants and antioxidants in the anti-inflammatory and apoptotic effects of curcumin (diferuloymethane). Free Rad Biol Med 43, 568–580.CrossRefGoogle Scholar
  20. Singh, S. (2007). From exotic spice to modern medicine. Cell 130, 765–768.CrossRefGoogle Scholar
  21. Singh, S., and Aggarwal, B. B. (1995). Activation of transcription factor NF-κB is suppressed by curcumin (diferuloylmethane). J Biol Chem. 2270, 24995–25000.CrossRefGoogle Scholar
  22. Sokoloski, J.A., Hodnick, W.F., Mayne, S.T., Cinquina, C., Kim, C.S., and Sartorelli, A.C. (1997). Induction of differentiation of HL-60 promyelocytic leukemia cells by vitamin E and other antioxidants in combination with low levels of low levels of vitamin D3: Possible relationship to NF-kappa B. Leukemia 11, 1546–1553.CrossRefGoogle Scholar
  23. Thaloor, D., Singh, A.K., Sidhu, G.S., Prasad, P.V., Kleinman, H.K. and Maheshwari, R.K. (1998). Inhibition of antigenic differentiation of human umbilical vein endothelial cells by curcumin. Cell Growth Differ 9, 305–312.Google Scholar
  24. Thaloor, D., Miller, K.J., Gephart, J., Mitchell, P.O., and Pavlath, G.K. (1999). Systemic administration of the NF-kappa B inhibitor curcumin stimulates muscle regeneration after traumatic injury. Amer J Physiol 277, C320–C329.Google Scholar
  25. Xu, P-H., Long, Y., Dai, F. and Liu, Z-L. (2007). The relaxant effect of curcumin on porcine coronary arterial ring segments. Vascular Biol 47, 25–30.Google Scholar
  26. Yang, X. (2005). A new role of p53 in maintaining genetic stability in embryonic stem cells. Cell Cycle 4, 363–364.CrossRefGoogle Scholar
  27. Zhu, D.Y., and Lou, Y.J. (2006). Icariin-mediated expression of cardiac genes and modulation of nitric oxide signaling pathway during differentiation of mouse embryonic stem cells into cardiomyocytes in vitro. Acta Pharmacol Sin 27, 311–320.CrossRefGoogle Scholar

Copyright information

© Higher Education Press and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Kalpana Mujoo
    • 1
    Email author
  • Lubov E. Nikonoff
    • 1
  • Vladislav G Sharin
    • 2
  • Nathan S. Bryan
    • 1
  • Alexander Y. Kots
    • 3
  • Ferid Murad
    • 3
  1. 1.Texas Therapeutics Institute, Brown Foundation Institute of Molecular MedicineUniversity of Texas Health Science Center at HoustonHoustonUSA
  2. 2.Departments of Immunology and PathologyBaylor College of MedicineHoustonUSA
  3. 3.Departments of Biochemistry and Molecular BiologyGeorge Washington UniversityWashingtonUSA

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