Alleviation of hepatic fibrosis and autophagy via inhibition of transforming growth factor-β1/Smads pathway through shikonin
Tong Liu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
These authors contributed equally to this work and shared first authorship.Search for more papers by this authorLing Xu
Department of Gastroenterology, Shanghai Tongren Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
These authors contributed equally to this work and shared first authorship.Search for more papers by this authorChengfen Wang
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorKan Chen
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorYujing Xia
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorJingjing Li
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorSainan Li
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorLiwei Wu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorJiao Feng
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorShizan Xu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Shanghai Tenth Hospital, School of Clinical Medicine of Nanjing Medical University, Shanghai, China
Search for more papers by this authorWenwen Wang
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorXiya Lu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorXiaoming Fan
Department of Gastroenterology, Jinshan Hospital of Fudan University, Shanghai, China
Search for more papers by this authorWenhui Mo
Department of Gastroenterology, Minhang Hospital, Fudan University, Shanghai, China
Search for more papers by this authorYingqun Zhou
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorCorresponding Author
Yan Zhao
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Yan Zhao and Chuanyong Guo, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China.
Email: [email protected]; [email protected]
Search for more papers by this authorCorresponding Author
Chuanyong Guo
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Yan Zhao and Chuanyong Guo, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China.
Email: [email protected]; [email protected]
Search for more papers by this authorTong Liu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
These authors contributed equally to this work and shared first authorship.Search for more papers by this authorLing Xu
Department of Gastroenterology, Shanghai Tongren Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
These authors contributed equally to this work and shared first authorship.Search for more papers by this authorChengfen Wang
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorKan Chen
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorYujing Xia
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorJingjing Li
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorSainan Li
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorLiwei Wu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorJiao Feng
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorShizan Xu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Shanghai Tenth Hospital, School of Clinical Medicine of Nanjing Medical University, Shanghai, China
Search for more papers by this authorWenwen Wang
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorXiya Lu
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorXiaoming Fan
Department of Gastroenterology, Jinshan Hospital of Fudan University, Shanghai, China
Search for more papers by this authorWenhui Mo
Department of Gastroenterology, Minhang Hospital, Fudan University, Shanghai, China
Search for more papers by this authorYingqun Zhou
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorCorresponding Author
Yan Zhao
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Yan Zhao and Chuanyong Guo, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China.
Email: [email protected]; [email protected]
Search for more papers by this authorCorresponding Author
Chuanyong Guo
Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Yan Zhao and Chuanyong Guo, Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, China.
Email: [email protected]; [email protected]
Search for more papers by this authorAbstract
Background and Aim
Liver fibrosis is a worldwide clinical challenge during the progression of chronic liver disease to liver cirrhosis. Shikonin is extracted from the root of Lithospermum erythrorhizon with antioxidant, anti-inflammatory, anticancer, and wound-healing properties. The study aims to investigate the protective effect of shikonin on liver fibrosis and its underlying mechanism.
Methods
Two liver fibrosis models were established in male C57 mice by intraperitoneal injection of CCl4 or bile duct ligation. Shikonin was administered orally three times weekly at a dose of 2.5 or 5 mg/kg. Protein and mRNA expressions were assayed by quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemical staining.
Results
Shikonin significantly inhibited activation of hepatic stellate cells and extracellular matrix formation by downregulating the transforming growth factor-β1 expression and maintaining the normal balance between metalloproteinase-2 and tissue inhibitor of metalloproteinase-1. Shikonin also decreased hepatic stellate cell energy production by inhibiting autophagy.
Conclusions
The results confirmed that shikonin attenuated liver fibrosis by downregulating the transforming growth factor-β1/Smads pathway and inhibiting autophagy.
References
- 1Dranoff JA, Wells RG. Portal fibroblasts: underappreciated mediators of biliary fibrosis. Hepatology 2010; 51: 1438–1444.
- 2Friedman SL. Hepatic fibrosis—overview. Toxicology 2008; 254: 120–129.
- 3Lim YS, Kim WR. The global impact of hepatic fibrosis and end-stage liver disease. Clin. Liver Dis. 2008; 12: 733–746.
- 4Sherman M, Klein A. AASLD single-topic research conference on hepatocellular carcinoma: conference proceedings. Hepatology 2004; 40: 1465–1473.
- 5Kisseleva T, Brenner DA. Role of hepatic stellate cells in fibrogenesis and the reversal of fibrosis. J. Gastroenterol. Hepatol. 2007; 22: S73–S78.
- 6De Minicis S, Seki E, Uchinami H et al. Gene expression profiles during hepatic stellate cell activation in culture and in vivo. Gastroenterology 2007; 132: 1937–1946.
- 7Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology 2008; 134: 1655–1669.
- 8Reeves HL, Friedman SL. Activation of hepatic stellate cells—a key issue in liver fibrosis. Front. Biosci. 2002; 7: d808–d826.
- 9Duval F, Moreno-Cuevas JE, Gonzalez-Garza MT, Rodriguez-Montalvo C, Cruz-Vega DE. Protective mechanisms of medicinal plants targeting hepatic stellate cell activation and extracellular matrix deposition in liver fibrosis. Chin. Med. 2014; 9: 27.
- 10Baghy K, Iozzo RV, Kovalszky I. Decorin–TGFβ axis in hepatic fibrosis and cirrhosis. J. Histochem. Cytochem. 2012; 60: 262–268.
- 11Meng XM, Nikolic-Paterson DJ, Lan HY. TGF-β: the master regulator of fibrosis. Nat. Rev. Nephrol. 2016; 12: 325–338.
- 12Shen M, Chen K, Lu J et al. Protective effect of astaxanthin on liver fibrosis through modulation of TGF-β1 expression and autophagy. Mediators Inflamm. 2014; 2014: 954502.
- 13Li J, Chen K, Li S et al. Protective effect of fucoidan from Fucus vesiculosus on liver fibrosis via the TGF-β1/Smad pathway-mediated inhibition of extracellular matrix and autophagy. Drug Des. Devel. Ther. 2016; 10: 619–630.
- 14Senties-Gomez MD, Galvez-Gastelum FJ, Meza-Garcia E, Armendariz-Borunda J. Hepatic fibrosis: role of matrix metalloproteases and TGFβ. Gac. Med. Mex. 2005; 141: 315–322.
- 15Ashford TP, Porter KR. Cytoplasmic components in hepatic cell lysosomes. J. Cell Biol. 1962; 12: 198–202.
- 16Zhou XJ, Zhang H. Autophagy in immunity: implications in etiology of autoimmune/autoinflammatory diseases. Autophagy 2012; 8: 1286–1299.
- 17Dong Y, Undyala VV, Gottlieb RA, Mentzer RJ, Przyklenk K. Autophagy: definition, molecular machinery, and potential role in myocardial ischemia–reperfusion injury. J. Cardiovasc. Pharmacol. Ther. 2010; 15: 220–230.
- 18Rautou PE, Mansouri A, Lebrec D, Durand F, Valla D, Moreau R. Autophagy in liver diseases. J. Hepatol. 2010; 53: 1123–1134.
- 19Kanayama M, Shinohara ML. Roles of autophagy and autophagy-related proteins in antifungal immunity. Front. Immunol. 2016; 7: 47.
- 20He Y, Zhu J, Huang Y, Gao H, Zhao Y. Advanced glycation end product (AGE)-induced hepatic stellate cell activation via autophagy contributes to hepatitis C-related fibrosis. Acta Diabetol. 2015; 52: 959–969.
- 21Thoen LF, Guimaraes EL, Grunsven LA. Autophagy: a new player in hepatic stellate cell activation. Autophagy 2012; 8: 126–128.
- 22Hernandez-Gea V, Ghiassi-Nejad Z, Rozenfeld R et al. Autophagy releases lipid that promotes fibrogenesis by activated hepatic stellate cells in mice and in human tissues. Gastroenterology 2012; 142: 938–946.
- 23Liu T, Xia Y, Li J et al. Shikonin attenuates concanavalin A-induced acute liver injury in mice via inhibition of the JNK pathway. Mediators Inflamm. 2016; 2016: 2748367.
- 24Liu T, Zhang Q, Mo W et al. The protective effects of shikonin on hepatic ischemia/reperfusion injury are mediated by the activation of the PI3K/Akt pathway. Sci. Rep. 2017; 7: 44785.
- 25Liang W, Cai A, Chen G et al. Shikonin induces mitochondria-mediated apoptosis and enhances chemotherapeutic sensitivity of gastric cancer through reactive oxygen species. Sci. Rep. 2016; 6: 38267.
- 26Fan C, Dong Y, Xie Y et al. Shikonin reduces TGF-β1-induced collagen production and contraction in hypertrophic scar-derived human skin fibroblasts. Int. J. Mol. Med. 2015; 36: 985–991.
- 27Han T, Zhang G, Yan D, Yang H, Ma T, Ye Z. Modulation of plasminogen activator inhibitor-1 (PAI-1) by the naphthoquinone shikonin. Fitoterapia 2016; 113: 117–122.
- 28Tsukada S, Parsons CJ, Rippe RA. Mechanisms of liver fibrosis. Clin. Chim. Acta 2006; 364: 33–60.
- 29Andujar I, Rios JL, Giner RM, Recio MC. Pharmacological properties of shikonin—a review of literature since 2002. Planta Med. 2013; 79: 1685–1697.
- 30Andujar I, Recio MC, Giner RM, Rios JL. Traditional Chinese medicine remedy to jury: the pharmacological basis for the use of shikonin as an anticancer therapy. Curr. Med. Chem. 2013; 20: 2892–2898.
- 31Wang R, Yin R, Zhou W, Xu D, Li S. Shikonin and its derivatives: a patent review. Expert Opin. Ther. Pat. 2012; 22: 977–997.
- 32Toosi AE. Liver fibrosis: causes and methods of assessment, a review. Rom. J. Intern. Med. 2015; 53: 304–314.
- 33Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story. Gut 2015; 64: 830–841.
- 34Zhou WC, Zhang QB, Qiao L. Pathogenesis of liver cirrhosis. World J. Gastroenterol. 2014; 20: 7312–7324.
- 35Thoen LF, Guimaraes EL, Dolle L et al. A role for autophagy during hepatic stellate cell activation. J. Hepatol. 2011; 55: 1353–1360.
- 36Li J, Wang F, Xia Y et al. Astaxanthin pretreatment attenuates hepatic ischemia reperfusion-induced apoptosis and autophagy via the ROS/MAPK pathway in mice. Mar. Drugs 2015; 13: 3368–3387.
- 37Shintani T, Klionsky DJ. Autophagy in health and disease: a double-edged sword. Science 2004; 306: 990–995.
- 38He W, Wang B, Yang J et al. Chloroquine improved carbon tetrachloride-induced liver fibrosis through its inhibition of the activation of hepatic stellate cells: role of autophagy. Biol. Pharm. Bull. 2014; 37: 1505–1509.
- 39Inagaki Y, Okazaki I. Emerging insights into transforming growth factor β Smad signal in hepatic fibrogenesis. Gut 2007; 56: 284–292.
- 40Meindl-Beinker NM, Dooley S. Transforming growth factor-β and hepatocyte transdifferentiation in liver fibrogenesis. J. Gastroenterol. Hepatol. 2008; 23: S122–S127.
- 41Ueberham E, Low R, Ueberham U, Schonig K, Bujard H, Gebhardt R. Conditional tetracycline-regulated expression of TGF-β1 in liver of transgenic mice leads to reversible intermediary fibrosis. Hepatology 2003; 37: 1067–1078.
- 42Williams AO, Knapton AD. Hepatic silicosis, cirrhosis, and liver tumors in mice and hamsters: studies of transforming growth factor β expression. Hepatology 1996; 23: 1268–1275.
- 43Gressner AM, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-β as major players and therapeutic targets. J. Cell. Mol. Med. 2006; 10: 76–99.
- 44Massague J, Seoane J, Wotton D. Smad transcription factors. Genes Dev. 2005; 19: 2783–2810.
- 45Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-β family signalling. Nature 2003; 425: 577–584.
- 46Pan CC, Kumar S, Shah N et al. Endoglin regulation of Smad2 function mediates Beclin1 expression and endothelial autophagy. J. Biol. Chem. 2015; 290: 14884–14892.
Citing Literature
