Selonsertib Inhibits Liver Fibrosis via Downregulation of ASK1/MAPK Pathway of Hepatic Stellate Cells

  • Yoon, Young-Chan (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Fang, Zhenghuan (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Lee, Ji Eun (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Park, Jung Hee (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Ryu, Ji-Kan (Department of Urology, College of Medicine, Inha University) ;
  • Jung, Kyung Hee (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Hong, Soon-Sun (Department of Biomedical Sciences, College of Medicine, and Program in Biomedical Science & Engineering, Inha University)
  • Received : 2020.02.05
  • Accepted : 2020.04.29
  • Published : 2020.11.01


Liver fibrosis constitutes a significant health problem worldwide due to its rapidly increasing prevalence and the absence of specific and effective treatments. Growing evidence suggests that apoptosis-signal regulating kinase 1 (ASK1) is activated in oxidative stress, which causes hepatic inflammation and apoptosis, leading to liver fibrogenesis through a mitogen-activated protein kinase (MAPK) downstream signals. In this study, we investigated whether selonsertib, a selective inhibitor of ASK1, shows therapeutic efficacy for liver fibrosis, and elucidated its mechanism of action in vivo and in vitro. As a result, selonsertib strongly suppressed the growth and proliferation of hepatic stellate cells (HSCs) and induced apoptosis by increasing Annexin V and TUNEL-positive cells. We also observed that selonsertib inhibited the ASK1/MAPK pathway, including p38 and c-Jun N-terminal kinase (JNK) in HSCs. Interestingly, dimethylnitrosamine (DMN)-induced liver fibrosis was significantly alleviated by selonsertib treatment in rats. Furthermore, selonsertib reduced collagen deposition and the expression of extracellular components such as α-smooth muscle actin (α-SMA), fibronectin, and collagen type I in vitro and in vivo. Taken together, selonsertib suppressed fibrotic response such as HSC proliferation and extracellular matrix components by blocking the ASK1/MAPK pathway. Therefore, we suggest that selonsertib may be an effective therapeutic drug for ameliorating liver fibrosis.



  1. Amos, L. A., Ma, F. Y., Tesch, G. H., Liles, J. T., Breckenridge, D. G., Nikolic-Paterson, D. J. and Han, Y. (2018) ASK1 inhibitor treatment suppresses p38/JNK signalling with reduced kidney inflammation and fibrosis in rat crescentic glomerulonephritis. J. Cell. Mol. Med. 22, 4522-4533.
  2. Arriazu, E., Ruiz de Galarreta, M., Cubero, F. J., Varela-Rey, M., Perez de Obanos, M. P., Leung, T. M., Lopategi, A., Benedicto, A., Abraham-Enachescu, I. and Nieto, N. (2014) Extracellular matrix and liver disease. Antioxid. Redox Signal. 21, 1078-1097.
  3. Baffy, G. (2013) Hepatocellular carcinoma in non-alcoholic fatty liver disease: epidemiology, pathogenesis, and prevention. J. Clin. Transl. Hepatol. 1, 131-137.
  4. Bataller, R. and Brenner, D. A. (2005) Liver fibrosis. J. Clin. Invest. 115, 209-218.
  5. Budas, G. R., Boehm, M., Kojonazarov, B., Viswanathan, G., Tian, X., Veeroju, S., Novoyatleva, T., Grimminger, F., Hinojosa-Kirschenbaum, F., Ghofrani, H. A., Weissmann, N., Seeger, W., Liles, J. T. and Schermuly, R. T. (2018) ASK1 inhibition halts disease progression in preclinical models of pulmonary arterial hypertension. Am. J. Respir. Crit. Care Med. 197, 373-385.
  6. Carpino, G., Morini, S., Ginanni Corradini, S., Franchitto, A., Merli, M., Siciliano, M., Gentili, F., Onetti Muda, A., Berloco, P., Rossi, M., Attili, A. F. and Gaudio, E. (2005) Alpha-SMA expression in hepatic stellate cells and quantitative analysis of hepatic fibrosis in cirrhosis and in recurrent chronic hepatitis after liver transplantation. Dig. Liver Dis. 37, 349-356.
  7. Cuenda, A. and Rousseau, S. (2007) p38 MAP-kinases pathway regulation, function and role in human diseases. Biochim. Biophys. Acta 1773, 1358-1375.
  8. De Oliveira da Silva, B., Ramos, L. F. and Moraes, K. C. M. (2017) Molecular interplays in hepatic stellate cells: apoptosis, senescence, and phenotype reversion as cellular connections that modulate liver fibrosis. Cell Biol. Int. 41, 946-959.
  9. Elsharkawy, A. M., Oakley, F. and Mann, D. A. (2005) The role and regulation of hepatic stellate cell apoptosis in reversal of liver fibrosis. Apoptosis 10, 927-939.
  10. Hatai, T., Matsuzawa, A., Inoshita, S., Mochida, Y., Kuroda, T., Sakamaki, K., Kuida, K., Yonehara, S., Ichijo, H. and Takeda, K. (2000) Execution of apoptosis signal-regulating kinase 1 (ASK1)-induced apoptosis by the mitochondria-dependent caspase activation. J. Biol. Chem. 275, 26576-26581.
  11. Hayakawa, R., Hayakawa, T., Takeda, K. and Ichijo, H. (2012) Therapeutic targets in the ASK1-dependent stress signaling pathways. Proc. Jpn. Acad. Ser. B Phys. Biol. Sci. 88, 434-453.
  12. Higashi, T., Friedman, S. L. and Hoshida, Y. (2017) Hepatic stellate cells as key target in liver fibrosis. Adv. Drug Deliv. Rev. 121, 27-42.
  13. Ichijo, H., Nishida, E., Irie, K., ten Dijke, P., Saitoh, M., Moriguchi, T., Takagi, M., Matsumoto, K., Miyazono, K. and Gotoh, Y. (1997) Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 275, 90-94.
  14. Ikejima, K., Takei, Y., Honda, H., Hirose, M., Yoshikawa, M., Zhang, Y. J., Lang, T., Fukuda, T., Yamashina, S., Kitamura, T. and Sato, N. (2002) Leptin receptor-mediated signaling regulates hepatic fibrogenesis and remodeling of extracellular matrix in the rat. Gastroenterology 122, 1399-1410.
  15. Kanda, T., Goto, T., Hirotsu, Y., Moriyama, M. and Omata, M. (2019) Molecular mechanisms driving progression of liver cirrhosis towards hepatocellular carcinoma in chronic hepatitis B and C infections: a review. Int. J. Mol. Sci. 20, 1358.
  16. Kanda, T., Matsuoka, S., Yamazaki, M., Shibata, T., Nirei, K., Takahashi, H., Kaneko, T., Fujisawa, M., Higuchi, T., Nakamura, H., Matsumoto, N., Yamagami, H., Ogawa, M., Imazu, H., Kuroda, K. and Moriyama, M. (2018) Apoptosis and non-alcoholic fatty liver diseases. World J. Gastroenterol. 24, 2661-2672.
  17. Kluwe, J., Pradere, J. P., Gwak, G. Y., Mencin, A., De Minicis, S., Osterreicher, C. H., Colmenero, J., Bataller, R. and Schwabe, R. F. (2010) Modulation of hepatic fibrosis by c-Jun-N-terminal kinase inhibition. Gastroenterology 138, 347-359.
  18. Kovalic, A. J., Satapathy, S. K. and Chalasani, N. (2018) Targeting incretin hormones and the ASK-1 pathway as therapeutic options in the treatment of non-alcoholic steatohepatitis. Hepatol. Int. 12, 97-106.
  19. Lee, J. H., Lee, H., Joung, Y. K., Jung, K. H., Choi, J. H., Lee, D. H., Park, K. D. and Hong, S. S. (2011) The use of low molecular weight heparin-pluronic nanogels to impede liver fibrosis by inhibition the TGF-beta/Smad signaling pathway. Biomaterials 32, 1438-1445.
  20. Liles, J. T., Corkey, B. K., Notte, G. T., Budas, G. R., Lansdon, E. B., Hinojosa-Kirschenbaum, F., Badal, S. S., Lee, M., Schultz, B. E., Wise, S., Pendem, S., Graupe, M., Castonguay, L., Koch, K. A., Wong, M. H., Papalia, G. A., French, D. M., Sullivan, T., Huntzicker, E. G., Ma, F. Y., Nikolic-Paterson, D. J., Altuhaifi, T., Yang, H., Fogo, A. B. and Breckenridge, D. G. (2018) ASK1 contributes to fibrosis and dysfunction in models of kidney disease. J. Clin. Invest. 128, 4485-4500.
  21. Loomba, R., Lawitz, E., Mantry, P. S., Jayakumar, S., Caldwell, S. H., Arnold, H., Diehl, A. M., Djedjos, C. S., Han, L., Myers, R. P., Subramanian, G. M., McHutchison, J. G., Goodman, Z. D., Afdhal, N. H. and Charlton M.R.; GS-US-384-1497 Investigators (2018) The ASK1 inhibitor selonsertib in patients with nonalcoholic steatohepatitis: a randomized, phase 2 trial. Hepatology 67, 549-559.
  22. Nakagawa, H., Maeda, S., Hikiba, Y., Ohmae, T., Shibata, W., Yanai, A., Sakamoto, K., Ogura, K., Noguchi, T., Karin, M., Ichijo, H. and Omata, M. (2008) Deletion of apoptosis signal-regulating kinase 1 attenuates acetaminophen-induced liver injury by inhibiting c-Jun N-terminal kinase activation. Gastroenterology 135, 1311-1321.
  23. Noureddin, M., Anstee, Q. M. and Loomba, R. (2016) Review article: emerging anti-fibrotic therapies in the treatment of non-alcoholic steatohepatitis. Aliment. Pharmacol. Ther. 43, 1109-1123.
  24. Pellicoro, A., Ramachandran, P., Iredale, J. P. and Fallowfield, J. A. (2014) Liver fibrosis and repair: immune regulation of wound healing in a solid organ. Nat. Rev. Immunol. 14, 181-194.
  25. Priya, S. and Sudhakaran, P. R. (2008) Cell survival, activation and apoptosis of hepatic stellate cells: modulation by extracellular matrix proteins. Hepatol. Res. 38, 1221-1232.
  26. Son, G., Hines, I. N., Lindquist, J., Schrum, L. W. and Rippe, R. A. (2009) Inhibition of phosphatidylinositol 3-kinase signaling in hepatic stellate cells blocks the progression of hepatic fibrosis. Hepatology 50, 1512-1523.
  27. Song, J., Cho, K. J., Cheon, S. Y., Kim, S. H., Park, K. A., Lee, W. T. and Lee, J. E. (2013) Apoptosis signal-regulating kinase 1 (ASK1) is linked to neural stem cell differentiation after ischemic brain injury. Exp. Mol. Med. 45, e69.
  28. Tobiume, K., Matsuzawa, A., Takahashi, T., Nishitoh, H., Morita, K., Takeda, K., Minowa, O., Miyazono, K., Noda, T. and Ichijo, H. (2001) ASK1 is required for sustained activations of JNK/p38 MAP kinases and apoptosis. EMBO Rep. 2, 222-228.
  29. Tsukada, S., Westwick, J. K., Ikejima, K., Sato, N. and Rippe, R. A. (2005) SMAD and p38 MAPK signaling pathways independently regulate alpha1(I) collagen gene expression in unstimulated and transforming growth factor-beta-stimulated hepatic stellate cells. J. Biol. Chem. 280, 10055-10064.
  30. Wang, Y., Gao, J., Zhang, D., Zhang, J., Ma, J. and Jiang, H. (2010) New insights into the antifibrotic effects of sorafenib on hepatic stellate cells and liver fibrosis. J. Hepatol. 53, 132-144.
  31. Yu, F. X., Teng, Y. Y., Zhu, Q. D., Zhang, Q. Y. and Tang, Y. H. (2014) Inhibitory effects of capsaicin on hepatic stellate cells and liver fibrosis. Biochem. Cell Biol. 92, 406-412.
  32. Zhang, C. Y., Yuan, W. G., He, P., Lei, J. H. and Wang, C. X. (2016) Liver fibrosis and hepatic stellate cells: etiology, pathological hallmarks and therapeutic targets. World J. Gastroenterol. 22, 10512-10522.
  33. Zhang, X. L., Chen, Z. N., Huang, Q. F., Bai, F. C., Nie, J. L., Lu, S. J., Wei, J. B. and Lin, X. (2018) Methyl helicterate inhibits hepatic stellate cell activation through modulation of apoptosis and autophagy. Cell. Physiol. Biochem. 51, 897-908.

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