Amygdalin inhibits HSC-T6 cell proliferation and fibrosis through the regulation of TGF-β/CTGF

Abstract

A mygdalin is one of the nitrilosides that was widely used to treat cancer, inhibit liver fibrosis. In the present study, the aim was to determine the antifibrotic potential of amygdalin and examine its mechanisms of action in vitro. Firstly, we found amygdalin significantly inhibited HSC-T6 cells proliferation. Both mRNA and protein of transforming growth factor-β (TGF-β) were decreased in HSC-T6 cells during amygdalin treatment. Secondly, to investigate functional role of TGF-β, both TGF-β over-expression vector and siRNA against TGF-β were transfected into HSC-T6 cells respectively. The results showed that over-expression of TGF-β promoted proliferation of HSC-T6 cells, whereas TGF-β knockdown inhibited cell viability. Moreover, our data even indicated that TGF-β could promote cell proliferation independent of amygdalin treatment. Finally, we found amygdalin could inhibit expression of the classical fibrotic factor αSMA, which suggested an antifibrotic effect of amygdalin. While the TGF-β antagonized anti-fibrotic effect of amygdalin. To assess the mechanisms, we examined expression of CTGF in cultured HSC-T6 cells. Our results showed that CTGF was down-regulated in HSCT6 cell treated by amygdalin, but was up-regulated when exogenous TGF-β introduced. As CTGF was one of the downstream factors in the TGF-β pathway. These might suggest that amygdalin inhibited HSC-T6 cells proliferation and fibrosis via TGF-β/CTGF pathway.

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References

  1. 1.

    Cho, A. et al. Detection of abnormally high amygdalin content in food by an enzyme immunoassay. Mol Cells 21:308–313 (2006).

    CAS  PubMed  Google Scholar 

  2. 2.

    Hwang, H. J. et al. Antinociceptive effect of amygdalin isolated from Prunus armeniaca on formalin-induced pain in rats. Biol Pharm Bull 31:1559–1564 (2008).

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Chang, H. K. et al. Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol Pharm Bull 29:1597–1602 (2006).

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Fukuda, T. et al. Anti-tumor promoting effect of glycosides from Prunus persica seeds. Biol Pharm Bull 26:271–300 (2003).

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Mirmiranpour, H. et al. Amygdalin inhibits angiogenesis in the cultured endothelial cells of diabetic rats. Indian J Pathol Microbiol 55:211–214 (2012).

    Article  PubMed  Google Scholar 

  6. 6.

    Lee, U. E. & Friedman, S. L. Mechanisms of hepatic fibrogenesis. Best Pract Res Clin Gastroenterol 25:195–206 (2011).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Inagaki, Y. & Okazaki, I. Emerging insights into Transforming growth factor beta Smad signal in hepatic fibrogenesis. Gut 56:284–292 (2007).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Limaye, P. B. et al. Mechanisms of hepatocyte growth factor-mediated and epidermal growth factor-mediated signaling in transdifferentiation of rat hepatocytes to biliary epithelium. Hepatology 47:1702–1713 (2008).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Date, M. et al. Modulation of transforming growth factor beta function in hepatocytes and hepatic stellate cells in rat liver injury. Gut 46:719–724 (2000).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. 10.

    Yoshida, K. & Matsuzaki, K. Differential Regulation of TGF-beta/Smad Signaling in Hepatic Stellate Cells between Acute and Chronic Liver Injuries. Front Physiol 3:53 (2012).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Yoshida, K. et al. Transforming growth factor-beta and platelet-derived growth factor signal via c-Jun N-terminal kinase-dependent Smad2/3 phosphorylation in rat hepatic stellate cells after acute liver injury. Am J Pathol 166:1029–1039 (2005).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Williams, E. J., Gaca, M. D., Brigstock, D. R., Arthur, M. J. & Benyon, R. C. Increased expression of connective tissue growth factor in fibrotic human liver and in activated hepatic stellate cells. J Hepatol 32:754–761 (2000).

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Huang, G. & Brigstock, D. R. Regulation of hepatic stellate cells by connective tissue growth factor. Front Biosci (Landmark Ed) 17:2495–2507 (2012).

    Article  Google Scholar 

  14. 14.

    Abou-Shady, M. et al. Connective tissue growth factor in human liver cirrhosis. Liver 20:296–304 (2000).

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Torok, N. J. Recent advances in the pathogenesis and diagnosis of liver fibrosis. J Gastroenterol 43:315–321 (2008).

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Puche, J. E., Saiman, Y. & Friedman, S. L. Hepatic stellate cells and liver fibrosis. Compr Physiol 3:1473–1492 (2013).

    Article  PubMed  Google Scholar 

  17. 17.

    Zhang, L. et al. Danshensu inhibits acetaldehyde-induced proliferation and activation of hepatic stellate cell-T6. Zhong Xi Yi Jie He Xue Ba 10:1155–1161 (2012).

    CAS  Article  Google Scholar 

  18. 18.

    Guo, J., Wu, W., Sheng, M., Yang, S. & Tan, J. Amygdalin inhibits renal fibrosis in chronic kidney disease. Mol Med Rep 7:1453–1457 (2013).

    CAS  PubMed  Google Scholar 

  19. 19.

    Xu, H. G. et al. Expression of ectonucleotide pyrophosphatase-1 in end-plate chondrocytes with transforming growth factor beta 1 siRNA interference by cyclic mechanical tension. Chin Med J 20:3886–3890 (2013).

    Google Scholar 

  20. 20.

    Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408 (2001).

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Wang, W. et al. Signaling mechanism of TGF-beta1 in prevention of renal inflammation:role of Smad7. J Am Soc Nephrol 16:1371–1383 (2005).

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Ha, M. H. et al. Effect of interferon-gamma on hepatic stellate cells stimulated by acetaldehyde. Hepatogastroenterology 84:1059–65 (2008).

    Google Scholar 

  23. 23.

    Hernandez-Gea, V. & Friedman, S. L. Pathogenesis of liver fibrosis. Annu Rev Pathol 6:425–456 (2011).

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Wells, R. G. The role of matrix stiffness in hepatic stellate cell activation and liver fibrosis. J Clin Gastroenterol 39:S158–161 (2005).

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Zardi, E. M. et al. New therapeutic approaches to liver fibrosis:a practicable route? Curr Med Chem 15:1628–1644 (2008).

    CAS  PubMed  Google Scholar 

  26. 26.

    Gressner, A. M. Cytokines and cellular crosstalk involved in the activation of fat-storing cells. J Hepatol 22:28–36 (1995).

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Lipson, K. E., Wong, C., Teng, Y. & Spong, S. CTGF is a central mediator of tissue remodeling and fibrosis and its inhibition can reverse the process of fibrosis. Fibrogenesis Tissue Repair 5:S24 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Gressner, O. A. & Gressner, A. M. Connective tissue growth factor:a fibrogenic master switch in fibrotic liver diseases. Liver Int 28:1065–1079 (2008).

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Friedman, S. L. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem 275:2247–2250 (2000).

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Potter, J. J. & Mezey, E. Acetaldehyde increases endogenous adiponectin and fibrogenesis in hepatic stellate cells but exogenous adiponectin inhibits fibrogenesis. Alcohol Clin Exp Res 31:2092–2100 (2007).

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Liu, Y. et al. Transforming growth factor-beta (TGFbeta)-mediated connective tissue growth factor (CTGF) expression in hepatic stellate cells requires Stat3 signaling activation. J Biol Chem 288:30708–30719 (2013).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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Correspondence to Xinhua Wang.

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Luo, H., Li, L., Tang, J. et al. Amygdalin inhibits HSC-T6 cell proliferation and fibrosis through the regulation of TGF-β/CTGF. Mol. Cell. Toxicol. 12, 265–271 (2016). https://doi.org/10.1007/s13273-016-0031-0

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Keywords

  • Amygdalin
  • HSC-T6 cells
  • Fibrosis
  • TGF-β/CTGF