Targeted blockade of JAK/STAT3 signaling inhibits proliferation, migration and collagen production as well as inducing the apoptosis of hepatic stellate cells

Gu,Yuan-Jing; Sun,Wu-Yi; Zhang,Sen; Li,Xin-Ran; Wei,Wei

doi:10.3892/ijmm.2016.2692

Targeted blockade of JAK/STAT3 signaling inhibits proliferation, migration and collagen production as well as inducing the apoptosis of hepatic stellate cells

Authors:
- Yuan-Jing Gu
- Wu-Yi Sun
- Sen Zhang
- Xin-Ran Li
- Wei Wei
View Affiliations

Affiliations: Institute of Clinical Pharmacology, Anhui Medical University, Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine, Hefei, Anhui 230032, P.R. China
Published online on: July 26, 2016 https://doi.org/10.3892/ijmm.2016.2692
Pages: 903-911

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Protein tyrosine kinases belonging to the Janus kinase (JAK) family are associated with many cytokine receptors, which, on ligand binding, regulate important cellular functions such as proliferation, apoptosis and differentiation. The protective effects of JAK inhibitors on fibrotic diseases such as myelofibrosis and bone marrow fibrosis have been demonstrated in previous studies. The JAK inhibitor SHR0302 is a synthetic molecule that potently inhibits all members of the JAK family, particularly JAK1. However, its effect on hepatic fibrosis has not been investigated to date, to the best of our knowledge. In the present study, the effects of SHR0302 on the activation, proliferation, migration and apoptosis of hepatic stellate cells (HSCs) as well as HSC collagen production were investigated. Our data demonstrated that treatment with SHR0302 (10-9-10-5 mol/l) exerted an inhibitory effect on the activation, proliferation and migration of HSCs. In addition, the expression of collagen I and collagen III were significantly decreased following treatment with SHR0302. Furthermore, SHR0302 induced the apoptosis of HSCs, which was demonstrated by Annexin V/PI staining. SHR0302 significantly increased the activation of caspase-3 and Bax in HSCs whereas it decreased the expression of Bcl-2. SHR0302 also inhibited the activation of Akt signaling pathway. The pharmacological inhibition of the JAK1/signal transducer and activator of transcription (STAT)3 pathway led to the disruption of functions essential for HSC growth. Taken together, these findings provide evidence that SHR0302 may have the potential to alleviate hepatic fibrosis by targeting HSC functions.

View Figures

View References

1	Leon DA and McCambridge J: Liver cirrhosis mortality rates in Britain from 1950 to 2002: an analysis of routine data. Lancet. 367:52–56. 2006. View Article : Google Scholar : PubMed/NCBI
2	Priya S and Sudhakaran PR: Cell survival, activation and apoptosis of hepatic stellate cells: modulation by extracellular matrix proteins. Hepatol Res. 38:1221–1232. 2008.PubMed/NCBI
3	Wang P, Liu T, Cong M, Wu X, Bai Y, Yin C, An W, Wang B, Jia J and You H: Expression of extracellular matrix genes in cultured hepatic oval cells: an origin of hepatic stellate cells through transforming growth factor beta? Liver Int. 29:575–584. 2009. View Article : Google Scholar : PubMed/NCBI
4	Moles A, Tarrats N, Morales A, Domínguez M, Bataller R, Caballería J, García-Ruiz C, Fernández-Checa JC and Marí M: Acidic sphingomyelinase controls hepatic stellate cell activation and in vivo liver fibrogenesis. Am J Pathol. 177:1214–1224. 2010. View Article : Google Scholar : PubMed/NCBI
5	Fallowfield JA: Therapeutic targets in liver fibrosis. Am J Physiol Gastrointest Liver Physiol. 300:G709–G715. 2011. View Article : Google Scholar : PubMed/NCBI
6	Povero D, Busletta C, Novo E, di Bonzo LV, Cannito S, Paternostro C and Parola M: Liver fibrosis: a dynamic and potentially reversible process. Histol Histopathol. 25:1075–1091. 2010.PubMed/NCBI
7	Seavey MM and Dobrzanski P: The many faces of Janus kinase. Biochem Pharmacol. 83:1136–1145. 2012. View Article : Google Scholar : PubMed/NCBI
8	Tanaka Y: Recent progress and perspective in JAK inhibitors for rheumatoid arthritis: from bench to bedside. J Biochem. 158:173–179. 2015. View Article : Google Scholar : PubMed/NCBI
9	Kisseleva T, Bhattacharya S, Braunstein J and Schindler CW: Signaling through the JAK/STAT pathway, recent advances and future challenges. Gene. 285:1–24. 2002. View Article : Google Scholar : PubMed/NCBI
10	Pardanani A, Vannucchi AM, Passamonti F, Cervantes F, Barbui T and Tefferi A: JAK inhibitor therapy for myelofibrosis: critical assessment of value and limitations. Leukemia. 25:218–225. 2011. View Article : Google Scholar
11	Johnson SJ, Hines JE and Burt AD: Phenotypic modulation of perisinusoidal cells following acute liver injury: a quantitative analysis. Int J Exp Pathol. 73:765–772. 1992.PubMed/NCBI
12	Cheng K, Yang N and Mahato RI: TGF-beta1 gene silencing for treating liver fibrosis. Mol Pharm. 6:772–779. 2009. View Article : Google Scholar : PubMed/NCBI
13	Kanzler S, Lohse AW, Keil A, Henninger J, Dienes HP, Schirmacher P, Rose-John S, zum Büschenfelde KH and Blessing M: TGF-beta1 in liver fibrosis: an inducible transgenic mouse model to study liver fibrogenesis. Am J Physiol. 276:G1059–G1068. 1999.PubMed/NCBI
14	Mair M, Blaas L, Österreicher CH, Casanova E and Eferl R: JAK-STAT signaling in hepatic fibrosis. Front Biosci (Landmark Ed). 16:2794–2811. 2011. View Article : Google Scholar
15	Paik YH, Kim JK, Lee JI, Kang SH, Kim DY, An SH, Lee SJ, Lee DK, Han KH, Chon CY, et al: Celecoxib induces hepatic stellate cell apoptosis through inhibition of Akt activation and suppresses hepatic fibrosis in rats. Gut. 58:1517–1527. 2009. View Article : Google Scholar : PubMed/NCBI
16	Son G, Hines IN, Lindquist J, Schrum LW and Rippe RA: Inhibition of phosphatidylinositol 3-kinase signaling in hepatic stellate cells blocks the progression of hepatic fibrosis. Hepatology. 50:1512–1523. 2009. View Article : Google Scholar : PubMed/NCBI
17	Schuppan D and Kim YO: Evolving therapies for liver fibrosis. J Clin Invest. 123:1887–1901. 2013. View Article : Google Scholar : PubMed/NCBI
18	Mallat A and Lotersztajn S: Cellular mechanisms of tissue fibrosis. 5. Novel insights into liver fibrosis. Am J Physiol Cell Physiol. 305:C789–C799. 2013. View Article : Google Scholar : PubMed/NCBI
19	Senoo H, Yoshikawa K, Morii M, Miura M, Imai K and Mezaki Y: Hepatic stellate cell (vitamin A-storing cell) and its relative-past, present and future. Cell Biol Int. 34:1247–1272. 2010. View Article : Google Scholar : PubMed/NCBI
20	Hernandez-Gea V and Friedman SL: Pathogenesis of liver fibrosis. Annu Rev Pathol. 6:425–456. 2011. View Article : Google Scholar
21	Choi JH, Hwang YP, Choi CY, Chung YC and Jeong HG: Anti-fibrotic effects of the anthocyanins isolated from the purple-fleshed sweet potato on hepatic fibrosis induced by dimethylnitrosamine administration in rats. Food Chem Toxicol. 48:3137–3143. 2010. View Article : Google Scholar : PubMed/NCBI
22	Clément S, Pascarella S, Conzelmann S, Gonelle-Gispert C, Guilloux K and Negro F: The hepatitis C virus core protein indirectly induces alpha-smooth muscle actin expression in hepatic stellate cells via interleukin-8. J Hepatol. 52:635–643. 2010. View Article : Google Scholar : PubMed/NCBI
23	Wu J and Zern MA: Hepatic stellate cells: a target for the treatment of liver fibrosis. J Gastroenterol. 35:665–672. 2000. View Article : Google Scholar : PubMed/NCBI
24	Pang M, Ma L, Gong R, Tolbert E, Mao H, Ponnusamy M, Chin YE, Yan H, Dworkin LD and Zhuang S: A novel STAT3 inhibitor, S3I-201, attenuates renal interstitial fibroblast activation and interstitial fibrosis in obstructive nephropathy. Kidney Int. 78:257–268. 2010. View Article : Google Scholar : PubMed/NCBI
25	Yu Y, Wang Y, Niu Y, Fu L, Chin YE and Yu C: Leukemia inhibitory factor attenuates renal fibrosis through Stat3-miR-29c. Am J Physiol Renal Physiol. 309:F595–F603. 2015. View Article : Google Scholar : PubMed/NCBI
26	Zhang W, Niu M, Yan K, Zhai X, Zhou Q, Zhang L and Zhou Y: Stat3 pathway correlates with the roles of leptin in mouse liver fibrosis and sterol regulatory element binding protein-1c expression of rat hepatic stellate cells. Int J Biochem Cell Biol. 45:736–744. 2013. View Article : Google Scholar : PubMed/NCBI
27	Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT and Horwitz AR: Cell migration: integrating signals from front to back. Science. 302:1704–1709. 2003. View Article : Google Scholar : PubMed/NCBI
28	Michaud-Levesque J, Bousquet-Gagnon N and Béliveau R: Quercetin abrogates IL-6/STAT3 signaling and inhibits glioblastoma cell line growth and migration. Exp Cell Res. 318:925–935. 2012. View Article : Google Scholar : PubMed/NCBI
29	Lim CP, Phan TT, Lim IJ and Cao X: Stat3 contributes to keloid pathogenesis via promoting collagen production, cell proliferation and migration. Oncogene. 25:5416–5425. 2006. View Article : Google Scholar : PubMed/NCBI
30	Schuppan D: Structure of the extracellular matrix in normal and fibrotic liver: collagens and glycoproteins. Semin Liver Dis. 10:1–10. 1990. View Article : Google Scholar : PubMed/NCBI
31	Rojkind M and Martinez-Palomo A: Increase in type I and type III collagens in human alcoholic liver cirrhosis. Proc Natl Acad Sci USA. 73:539–543. 1976. View Article : Google Scholar : PubMed/NCBI
32	Grimaud JA, Druguet M, Peyrol S, Chevalier O, Herbage D and El Badrawy N: Collagen immunotyping in human liver: light and electron microscope study. J Histochem Cytochem. 28:1145–1156. 1980. View Article : Google Scholar : PubMed/NCBI
33	Lakner AM, Moore CC, Gulledge AA and Schrum LW: Daily genetic profiling indicates JAK/STAT signaling promotes early hepatic stellate cell transdifferentiation. World J Gastroenterol. 16:5047–5056. 2010. View Article : Google Scholar : PubMed/NCBI
34	Hellerbrand C, Stefanovic B, Giordano F, Burchardt ER and Brenner DA: The role of TGFbeta1 in initiating hepatic stellate cell activation in vivo. J Hepatol. 30:77–87. 1999. View Article : Google Scholar : PubMed/NCBI
35	Thompson HJ, Strange R and Schedin PJ: Apoptosis in the genesis and prevention of cancer. Cancer Epidemiol Biomarkers Prev. 1:597–602. 1992.PubMed/NCBI
36	Bataller R and Brenner DA: Hepatic stellate cells as a target for the treatment of liver fibrosis. Semin Liver Dis. 21:437–451. 2001. View Article : Google Scholar : PubMed/NCBI
37	Zhao W, Su W, Kuang P, Zhang L, Liu J, Yin Z and Wang X: The role of hepatic stellate cells in the regulation of T-cell function and the promotion of hepatocellular carcinoma. Int J Oncol. 41:457–464. 2012.PubMed/NCBI
38	Kawada N: Human hepatic stellate cells are resistant to apoptosis: implications for human fibrogenic liver disease. Gut. 55:1073–1074. 2006. View Article : Google Scholar : PubMed/NCBI
39	Novo E, Marra F, Zamara E, Valfrè di Bonzo L, Monitillo L, Cannito S, Petrai I, Mazzocca A, Bonacchi A, De Franco RS, et al: Overexpression of Bcl-2 by activated human hepatic stellate cells: resistance to apoptosis as a mechanism of progressive hepatic fibrogenesis in humans. Gut. 55:1174–1182. 2006. View Article : Google Scholar : PubMed/NCBI
40	Nielsen M, Kaestel CG, Eriksen KW, Woetmann A, Stokkedal T, Kaltoft K, Geisler C, Röpke C and Odum N: Inhibition of constitutively activated Stat3 correlates with altered Bcl-2/Bax expression and induction of apoptosis in mycosis fungoides tumor cells. Leukemia. 13:735–738. 1999. View Article : Google Scholar : PubMed/NCBI
41	Lee SY, Kwok SK, Son HJ, Ryu JG, Kim EK, Oh HJ, Cho ML, Ju JH, Park SH and Kim HY: IL-17-mediated Bcl-2 expression regulates survival of fibroblast-like synoviocytes in rheumatoid arthritis through STAT3 activation. Arthritis Res Ther. 15:R312013. View Article : Google Scholar : PubMed/NCBI
42	Moodley YP, Misso NL, Scaffidi AK, Fogel-Petrovic M, McAnulty RJ, Laurent GJ, Thompson PJ and Knight DA: Inverse effects of interleukin-6 on apoptosis of fibroblasts from pulmonary fibrosis and normal lungs. Am J Respir Cell Mol Biol. 29:490–498. 2003. View Article : Google Scholar : PubMed/NCBI
43	Galluzzi L, Kepp O and Kroemer G: Caspase-3 and prostaglandins signal for tumor regrowth in cancer therapy. Oncogene. 31:2805–2808. 2012. View Article : Google Scholar
44	Yang B, El Nahas AM, Thomas GL, Haylor JL, Watson PF, Wagner B and Johnson TS: Caspase-3 and apoptosis in experimental chronic renal scarring. Kidney Int. 60:1765–1776. 2001. View Article : Google Scholar : PubMed/NCBI
45	Zhou TB, Qin YH, Zhou C, Lei FY, Zhao YJ, Chen J, Su LN and Huang WF: Less expression of prohibitin is associated with increased caspase-3 expression and cell apoptosis in renal interstitial fibrosis rats. Nephrology (Carlton). 17:189–196. 2012. View Article : Google Scholar
46	Jiang JX, Mikami K, Venugopal S, Li Y and Török NJ: Apoptotic body engulfment by hepatic stellate cells promotes their survival by the JAK/STAT and Akt/NF-kappaB-dependent pathways. J Hepatol. 51:139–148. 2009. View Article : Google Scholar : PubMed/NCBI
47	O'Sullivan LA, Liongue C, Lewis RS, Stephenson SE and Ward AC: Cytokine receptor signaling through the Jak-Stat-Socs pathway in disease. Mol Immunol. 44:2497–2506. 2007. View Article : Google Scholar : PubMed/NCBI
48	Xu Q, Briggs J, Park S, Niu G, Kortylewski M, Zhang S, Gritsko T, Turkson J, Kay H, Semenza GL, et al: Targeting Stat3 blocks both HIF-1 and VEGF expression induced by multiple oncogenic growth signaling pathways. Oncogene. 24:5552–5560. 2005. View Article : Google Scholar : PubMed/NCBI
49	Saxena NK, Sharma D, Ding X, Lin S, Marra F, Merlin D and Anania FA: Concomitant activation of the JAK/STAT, PI3K/AKT, and ERK signaling is involved in leptin-mediated promotion of invasion and migration of hepatocellular carcinoma cells. Cancer Res. 67:2497–2507. 2007. View Article : Google Scholar : PubMed/NCBI
50	Niu L, Wang X, Li J, Huang Y, Yang Z, Chen F, Ni H, Jin Y, Lu X and Cao Q: Leptin stimulates alpha1(I) collagen expression in human hepatic stellate cells via the phosphatidylinositol 3-kinase/Akt signalling pathway. Liver Int. 27:1265–1272. 2007. View Article : Google Scholar : PubMed/NCBI
51	Monaghan KA, Khong T, Burns CJ and Spencer A: The novel JAK inhibitor CYT387 suppresses multiple signalling pathways, prevents proliferation and induces apoptosis in phenotypically diverse myeloma cells. Leukemia. 25:1891–1899. 2011. View Article : Google Scholar : PubMed/NCBI
52	Gross ER, Hsu AK and Gross GJ: The JAK/STAT pathway is essential for opioid-induced cardioprotection: JAK2 as a mediator of STAT3, Akt, and GSK-3 beta. Am J Physiol Heart Circ Physiol. 291:H827–H834. 2006. View Article : Google Scholar : PubMed/NCBI
53	Tanaka Y and Yamaoka K: JAK inhibitor tofacitinib for treating rheumatoid arthritis: from basic to clinical. Mod Rheumatol. 23:415–424. 2013. View Article : Google Scholar
54	An HJ, Choi EK, Kim JS, Hong SW, Moon JH, Shin JS, Ha SH, Kim KP, Hong YS, Lee JL, et al: INCB018424 induces apoptotic cell death through the suppression of pJAK1 in human colon cancer cells. Neoplasma. 61:56–62. 2014. View Article : Google Scholar
55	Suryani S, Bracken LS, Harvey RC, Sia KC, Carol H, Chen IM, Evans K, Dietrich PA, Roberts KG, Kurmasheva RT, et al: Evaluation of the in vitro and in vivo efficacy of the JAK inhibitor AZD1480 against JAK-mutated acute lymphoblastic leukemia. Mol Cancer Ther. 14:364–374. 2015. View Article : Google Scholar :
56	Swaim SJ: Ruxolitinib for the treatment of primary myelofibrosis. Am J Health Syst Pharm. 71:453–462. 2014. View Article : Google Scholar : PubMed/NCBI
57	Ostojic A, Vrhovac R and Verstovsek S: Ruxolitinib for the treatment of myelofibrosis: its clinical potential. Ther Clin Risk Manag. 8:95–103. 2012.PubMed/NCBI
58	Cao Q, Mak KM and Lieber CS: Leptin enhances alpha1(I) collagen gene expression in LX-2 human hepatic stellate cells through JAK-mediated H₂O₂-dependent MAPK pathways. J Cell Biochem. 97:188–197. 2006. View Article : Google Scholar