Article | Published:

BMP-2 restoration aids in recovery from liver fibrosis by attenuating TGF-β1 signaling

Laboratory Investigationvolume 98pages9991013 (2018) | Download Citation


Transforming growth factor-β (TGF-β) plays a central role in hepatic fibrogenesis. This study investigated the function and mechanism of bone morphogenetic protein-2 (BMP-2) in regulation of hepatic fibrogenesis. BMP-2 expression in fibrotic liver was measured in human tissue microarray and mouse models of liver fibrosis induced by bile duct ligation surgery or carbon tetrachloride administration. Adenovirus-mediated BMP-2 gene delivery was used to test the prophylactic effect on liver fibrosis. Primary hepatic stellate cells (HSC), HSC-T6 and clone-9 cell lines were used to study the interplay between BMP-2 and TGF-β1. Hepatic BMP-2 was localized in parenchymal hepatocytes and activated HSCs and significantly decreased in human and mouse fibrotic livers, showing an opposite pattern of hepatic TGF-β1 contents. BMP-2 gene delivery alleviated the elevations of serum hepatic enzymes, cholangiocyte marker CK19, HSC activation markers, and liver fibrosis in both models. Mechanistically, exogenous TGF-β1 dose dependently reduced BMP-2 expression, whereas BMP-2 significantly suppressed expression of TGF-β and its cognate type I and II receptor peptides, as well as the induced Smad3 phosphorylation levels in primary mouse HSCs. Aside from its suppressive effects on cell proliferation and migration, BMP-2 treatment prominently attenuated the TGF-β1-stimulated α-SMA and fibronectin expression, and reversed the TGF-β1-modulated epithelial-to-mesenchymal transition marker expression in mouse HSCs. The mutual regulation between BMP-2 and TGF-β1 signaling axes may constitute the anti-fibrogenic mechanism of BMP-2 in the pathogenesis of liver fibrosis. BMP-2 may potentially serve as a novel therapeutic target for treatment of liver fibrosis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Additional information

These authors contributed equally: Yueh-Hua Chung, Ying-Hsien Huang.


  1. 1.

    Lee UE, Friedman SL. Mechanisms of hepatic fibrogenesis. Best Pract Res Clin Gastroenterol. 2011;25:195–206.

  2. 2.

    Friedman SL. Evolving challenges in hepatic fibrosis. Nat Rev Gastroenterol Hepatol. 2010;7:425–36.

  3. 3.

    Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005;115:209–18.

  4. 4.

    Modol T, Brice N, Ruiz de Galarreta M, et al. Fibronectin peptides as potential regulators of hepatic fibrosis through apoptosis of hepatic stellate cells. J Cell Physiol. 2015;230:546–53.

  5. 5.

    Liu XY, Liu RX, Hou F, et al. Fibronectin expression is critical for liver fibrogenesis in vivo and in vitro. Mol Med Rep. 2016;14:3669–75.

  6. 6.

    Bissell DM, Roulot D, George J. Transforming growth factor beta and the liver. Hepatology. 2001;34:859–67.

  7. 7.

    Jonsson JR, Clouston AD, Ando Y, et al. Angiotensin-converting enzyme inhibition attenuates the progression of rat hepatic fibrosis. Gastroenterology. 2001;121:148–55.

  8. 8.

    O’Connor JW, Gomez EW. Biomechanics of TGFbeta-induced epithelial-mesenchymal transition: implications for fibrosis and cancer. Clin Transl Med. 2014;3:23.

  9. 9.

    Bottinger EP. TGF-beta in renal injury and disease. Semin Nephrol. 2007;27:309–20.

  10. 10.

    Liu X, Hu H, Yin JQ. Therapeutic strategies against TGF-beta signaling pathway in hepatic fibrosis. Liver Int. 2006;26:8–22.

  11. 11.

    Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat Med. 2003;9:964–8.

  12. 12.

    Park J, Schwarzbauer JE. Mammary epithelial cell interactions with fibronectin stimulate epithelial-mesenchymal transition. Oncogene. 2014;33:1649–57.

  13. 13.

    Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med. 1994;331:1286–92.

  14. 14.

    Wozney JM, Rosen V, Celeste AJ, et al. Novel regulators of bone formation: molecular clones and activities. Science. 1988;242:1528–34.

  15. 15.

    Massague J. The transforming growth factor-beta family. Annu Rev Cell Biol. 1990;6:597–641.

  16. 16.

    Itoh S, Itoh F, Goumans MJ, et al. Signaling of transforming growth factor-beta family members through Smad proteins. Eur J Biochem. 2000;267:6954–67.

  17. 17.

    Feng XH, Derynck R. Specificity and versatility in TGF-beta signaling through Smads. Annu Rev Cell Dev Biol. 2005;21:659–93.

  18. 18.

    Eickelberg O, Morty RE. Transforming growth factor beta/bone morphogenic protein signaling in pulmonary arterial hypertension: remodeling revisited. Trends Cardiovasc Med. 2007;17:263–9.

  19. 19.

    Song JJ, Celeste AJ, Kong FM, et al. Bone morphogenetic protein-9 binds to liver cells and stimulates proliferation. Endocrinology. 1995;136:4293–7.

  20. 20.

    Duncan SA, Watt AJ. BMPs on the road to hepatogenesis. Genes Dev. 2001;15:1879–84.

  21. 21.

    Tsai MS, Suksaweang S, Jiang TX, et al. Proper BMP signaling levels are essential for 3D assembly of hepatic cords from hepatoblasts and mesenchymal cells. Dig Dis Sci. 2015;60:3669–80.

  22. 22.

    Xu CP, Ji WM, van den Brink GR, et al. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver. World J Gastroenterol. 2006;12:7621–5.

  23. 23.

    Yang YL, Liu YS, Chuang LY, et al. Bone morphogenetic protein-2 antagonizes renal interstitial fibrosis by promoting catabolism of type I transforming growth factor-beta receptors. Endocrinology. 2009;150:727–40.

  24. 24.

    Yang YL, Ju HZ, Liu SF, et al. BMP-2 suppresses renal interstitial fibrosis by regulating epithelial-mesenchymal transition. J Cell Biochem. 2011;112:2558–65.

  25. 25.

    Shlyonsky V, Soussia IB, Naeije R, et al. Opposing effects of bone morphogenetic protein-2 and endothelin-1 on lung fibroblast chloride currents. Am J Respir Cell Mol Biol. 2011;45:1154–60.

  26. 26.

    Gao X, Cao Y, Staloch DA, et al. Bone morphogenetic protein signaling protects against cerulein-induced pancreatic fibrosis. PLoS ONE. 2014;9:e89114.

  27. 27.

    Kao YH, Chen CL, Jawan B, et al. Upregulation of hepatoma-derived growth factor is involved in murine hepatic fibrogenesis. J Hepatol. 2010;52:96–105.

  28. 28.

    Massoner P, Kugler KG, Unterberger K, et al. Characterization of transcriptional changes in ERG rearrangement-positive prostate cancer identifies the regulation of metabolic sensors such as neuropeptide Y. PLoS ONE. 2013;8:e55207.

  29. 29.

    Tai MH, Cheng H, Wu JP, et al. Gene transfer of glial cell line-derived neurotrophic factor promotes functional recovery following spinal cord contusion. Exp Neurol. 2003;183:508–15.

  30. 30.

    Huang YH, Tiao MM, Huang LT, et al. Activation of miR-29a in activated hepatic stellate cells modulates its profibrogenic phenotype through inhibition of histone deacetylases 4. PLoS ONE. 2015;10:e0136453.

  31. 31.

    Kao YH, Chen PH, Wu TY, et al. Lipopolysaccharides induce Smad2 phosphorylation through PI3K/Akt and MAPK cascades in HSC-T6 hepatic stellate cells. Life Sci. 2017;184:37–46.

  32. 32.

    Yang YL, Wang FS, Li SC, et al. MicroRNA-29a alleviates bile duct ligation exacerbation of hepatic fibrosis in mice through epigenetic control of methyltransferases. Int J Mol Sci. 2017;18:192.

  33. 33.

    Nakatsuka R, Taniguchi M, Hirata M, et al. Transient expression of bone morphogenic protein-2 in acute liver injury by carbon tetrachloride. J Biochem. 2007;141:113–9.

  34. 34.

    Cai J, Zhao Y, Liu Y, et al. Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology. 2007;45:1229–39.

  35. 35.

    Li X, Yuan J, Li W, et al. Direct differentiation of homogeneous human adipose stem cells into functional hepatocytes by mimicking liver embryogenesis. J Cell Physiol. 2014;229:801–12.

  36. 36.

    Yu Q, Stamenkovic I. Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumor invasion and angiogenesis. Genes Dev. 2000;14:163–76.

  37. 37.

    Kao YH, Jawan B, Goto S, et al. Serum factors potentiate hypoxia-induced hepatotoxicity in vitro through increasing transforming growth factor-beta1 activation and release. Cytokine. 2009;47:11–22.

  38. 38.

    Han YP, Yan C, Zhou L, et al. A matrix metalloproteinase-9 activation cascade by hepatic stellate cells in trans-differentiation in the three-dimensional extracellular matrix. J Biol Chem. 2007;282:12928–39.

  39. 39.

    Choi YA, Kang SS, Jin EJ. BMP-2 treatment of C3H10T1/2 mesenchymal cells blocks MMP-9 activity during chondrocyte commitment. Cell Biol Int. 2009;33:887–92.

  40. 40.

    Zhang QD, Xu MY, Cai XB, et al. Myofibroblastic transformation of rat hepatic stellate cells: the role of Notch signaling and epithelial-mesenchymal transition regulation. Eur Rev Med Pharmacol Sci. 2015;19:4130–8.

  41. 41.

    Gao X, Cao Y, Yang W, et al. BMP-2 inhibits TGF-beta-induced pancreatic stellate cell activation and extracellular matrix formation. Am J Physiol Gastrointest Liver Physiol. 2013;304:G804–813.

Download references


This study was supported by the grants in part from Kaohsiung Chang Gung Memorial Hospital (Nos. CMRPG8F1561, CMRPG8F1562) and Kaohsiung Armed Forces General Hospital (No. 103-6), Taiwan.

Author information

Author notes


    1. Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Yueh-Hua Chung
      • , Shih-Chung Huang
      •  & Ming-Hong Tai
    2. Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital and Chiayi Chang Gung Memorial Hospital, Puzi City, Taiwan

      • Ying-Hsien Huang
    3. Center for Neuroscience, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Tien-Huei Chu
      •  & Ming-Hong Tai
    4. Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan

      • Chun-Lin Chen
    5. Division of Hepato-Gastroenterology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

      • Pey-Ru Lin
      •  & Tsung-Hui Hu
    6. Department of Internal Medicine, Kaohsiung Armed Forces General Hospital, Kaohsiung, Taiwan

      • Shih-Chung Huang
    7. Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung, Taiwan

      • Deng-Chyang Wu
    8. Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

      • Deng-Chyang Wu
    9. Biobank and Tissue Bank and Department of Pathology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

      • Chao-Cheng Huang
    10. Department of Medical Research, E-Da Hospital, Kaohsiung, Taiwan

      • Ying-Hsien Kao


    1. Search for Yueh-Hua Chung in:

    2. Search for Ying-Hsien Huang in:

    3. Search for Tien-Huei Chu in:

    4. Search for Chun-Lin Chen in:

    5. Search for Pey-Ru Lin in:

    6. Search for Shih-Chung Huang in:

    7. Search for Deng-Chyang Wu in:

    8. Search for Chao-Cheng Huang in:

    9. Search for Tsung-Hui Hu in:

    10. Search for Ying-Hsien Kao in:

    11. Search for Ming-Hong Tai in:

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Corresponding authors

    Correspondence to Ying-Hsien Kao or Ming-Hong Tai.

    Electronic supplementary material

    About this article

    Publication history