Antifibrotic effects of gallic acid on hepatic stellate cells: In vitro and in vivo mechanistic study

10.1016/j.jtcme.2018.01.010

Naglaa M. El-Lakkany；Walaa H. El-Maadawy；Sayed H. Seif el-Din；Samira Saleh；Marwa M. Safar；Shahira M. Ezzat；Salwa H. Mohamed；Sanaa S. Botros；Zeinab Demerdash；Olfat A. Hammam

Gallic acid ； Hepatic stellate cells ； Hepatocytes ； Proliferation/activation ； Apoptosis

Journal of Traditional and Complementary Medicine

9卷1期（2019 / 01 / 01）

45 - 53

英文

Few studies reported the antifibrotic effects of gallic acid (GA) despite its known hepatoprotective and antioxidant activities. Accordingly, this study investigated the antifibrotic effects of GA through clarifying its mechanisms on hepatic stellate cells' (HSCs) activation, proliferation and/or apoptosis. In vitro effects of GA on HSC-T6 activation/proliferation, morphology and safety on hepatocytes were assessed. In vivo, hepatic fibrosis was induced via chronic thioacetamide (TAA)-intoxication. TAA-intoxicated rats were treated with silyamrin or GA. At end of experiment, liver functions, hepatic MDA, GSH, PDGF-BB, TGF-β1, TIMP-1 and hydroxyproline were determined. Histological analysis and Sirius red staining of hepatic sections, expressions of alpha-smooth muscle actin (α-SMA), proliferating cellular nuclear antigen (PCNA) and caspase-3 were examined. In vitro, GA resulted in a concentration and time-dependent inhibition in HSCs activation, proliferation (IC_(50)= 45 and 19 μg/mL at 24 and 48 h respectively); restored the quiescent morphology of some activated HSCs plus its safety on hepatocytes. In vivo, GA reduced ALT, AST, MDA, PDGF-BB levels, collagen deposition and fibrosis score (S1 vs S4); increased caspase-3 expression and restored GSH stores, TGF-β1 level, α-SMA and PCNA expressions. In conclusion, GA counteracted the progression of hepatic fibrosis through reduction of HSCs proliferation/activation mutually with their apoptosis induction.

1. Atta, HM,Al-Hendy, A,Salama, SA,Shaker, OG,Hammam, OA(2009).Low-dose simultaneous delivery of adenovirus encoding hepatocyte growth factor and vascular endothelial growth factor in dogs enhances liver proliferation without systemic growth factor elevation.Liver Int,29,1022-1030.
2. Basheer, L,Kerem, Z(2015).Interactions between CYP3A4 and dietary polyphenols.Oxid Med Cell Longev,2015,854015.
3. Bassiouny, AR,Zaky, AZ,Abdulmalek, SA,Kandeel, KM,Ismail, A,Moftah, M(2011).Modulation of AP-endonuclease1 levels associated with hepatic cirrhosis in rat model treated with human umbilical cord blood mononuclear stem cells.Int J Clin Exp Pathol,4,692-707.
4. Borkham-Kamphorst, E,Kovalenko, E,van, Roeyen, CR(2008).Platelet-derived growth factor isoform expression in carbon tetrachloride-induced chronic liver injury.Lab Invest,88,1090-1100.
5. Chang, YJ,Hsu, SL,Liu, YT(2015).Gallic acid induces necroptosis via TNF-a signaling pathway in activated hepatic stellate cells.PLoS One,10,e0120713.
6. Chen, SR,Chen, XP,Lu, JJ,Wang, Y,Wang, YT(2015).Potent natural products and herbal medicines for treating liver fibrosis.Chin Med,10,7.
7. Chuang, CY,Liu, HC,Wu, LC,Chen, CY,Chang, JT,Hsu, SL(2010).Gallic acid induces apoptosis of lung fibroblasts via a reactive oxygen species-dependent ataxia telangiectasia mutated-p53 activation pathway.J Agric Food Chem,58,2943-2951.
8. de la Lastra, CA,Villegas, I(2007).Resveratrol as an antioxidant and pro-oxidant agent: mechanisms and clinical implications..Biochem Soc Trans,35,1156-1160.
9. Di Lullo, GA,Sweeney, SM,Korkko, J,Ala-Kokko, L,San Antonio, JD(2002).Mapping the ligand-binding sites and disease-associated mutations on the most abundant protein in the human, type I collagen.J Biol Chem,277,4223-4231.
10. Elango, S,Balwas, R,Padma, VV(2011).Gallic acid isolated from pomegranate peel extract induces reactive oxygen species mediated apoptosis in A549 cell line.J Canc Ther,2,638-645.
11. Ellman, GL(1959).Tissue sulfhydryl groups.Arch Biochem Biophys,82,70-77.
12. Faried, A,Kurnia, D,Faried, LS(2007).Anticancer effects of gallic acid isolated from Indonesian herbal medicine, Phaleria macrocarpa (Scheff.) Boerl, on human cancer cell lines.Int J Oncol.,30,605-613.
13. Friedman, SL(2008).Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver.Physiol Rev,88,125-172.
14. Friedman, SL(2008).Mechanisms of hepatic fibrogenesis.Gastroenterology,134,1655-1669.
15. Gómez-Caravaca, AM,Verardo, V,Toselli, M,Segura-Carretero, A,Fernández-Gutierrez, A,Caboni, MF(2013).Determination of the major phenolic compounds in pomegranate juices by HPLCDADESI-MS.J Agric Food Chem,61,5328-5337.
16. Hagens, WI,Beljaars, L,Mann, DA(2008).Cellular targeting of the apoptosis-inducing compound gliotoxin to fibrotic rat livers.J Pharmacol Exp Therapeut,324,902-910.
17. Hauff, P,Gottwald, U,Ocker, M(2015).Early to Phase II drugs currently under investigation for the treatment of liver fibrosis.Expet Opin Invest Drugs,24,309-327.
18. Hong, M,Li, S,Tan, HY,Wang, N,Tsao, SW,Feng, Y(2015).Current status of herbal medicines in chronic liver disease therapy: the biological effects, molecular targets and future prospects.Int J Mol Sci.,16,28705-28745.
19. Hsieh, SC,Wu, CH,Wu, CC(2014).Gallic acid selectively induces the necrosis of activated hepatic stellate cells via a calcium-dependent calpain I activation pathway.Life Sci,102,55-64.
20. Inoue, M,Sakaguchi, N,Isuzugawa, K,Tani, H,Ogihara, Y(2000).Role of reactive oxygen species in gallic acid-induced apoptosis.Biol Pharm Bull,23,1153-1157.
21. Iredale, JP(2007).Models of liver fibrosis: exploring the dynamic nature of inflammation and repair in a solid organ.J Clin Invest,117,539-548.
22. Ishikawa, Y,Kitamura, M(1998).Unexpected suppression of alpha-smooth muscle actin, the activation marker of mesangial cells, by pp60v-src tyrosine kinase.Biochem Biophys Res Commun,244,806-811.
23. Jadon, A,Bhadauria, M,Shukla, S(2007).Protective effect of Terminalia belerica Roxb. and gallic acid against carbon tetrachloride induced damage in albino rats.J Ethnopharmacol,109,214-218.
24. Jancová, P,Anzenbacherová, E,Papousková, B(2007).Silybin is metabolized by cytochrome P450 2C8 in vitro.Drug Metab Dispos,35,2035-2039.
25. Kadir, FA,Kassim, NM,Abdulla, MA,Yehye, WA(2013).Hepatoprotective role of ethanolic extract of Vitex negundo in thioacetamide-induced liver fibrosis in male rats.Evid Based Complement Alternat Med,2013,739850.
26. Kang, N,Gores, GJ,Shah, VH(2011).Hepatic stellate cells: partners in crime for liver metastases?.Hepatology,54,707-713.
27. Kim, YJ(2007).Antimelanogenic and antioxidant properties of gallic acid.Biol Pharm Bull,30,1052-1055.
28. Kisseleva, T,Brenner, DA(2006).Hepatic stellate cells and the reversal of fibrosis.J Gastroenterol Hepatol,21(Suppl 1),S84-S87.
29. Lee, UE,Friedman, SL(2011).Mechanisms of hepatic fibrogenesis.Best Pract Res Clin Gastroenterol,25,195-206.
30. Li, JT,Liao, ZX,Ping, J,Xu, D,Wang, H(2008).Molecular mechanism of hepatic stellate cell activation and antifibrotic therapeutic strategies.J Gastroenterol,43,419-428.
31. Li, L,Hu, Z,Li, W(2012).Establishment of a standardized liver fibrosis model with different pathological stages in rats.Gastroenterol Res Pract,2012,560345.
32. Li, YF,Guo, CJ,Yang, JJ,Wei, JY,Xu, J,Cheng, S(2006).Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract.Food Chem,96,254-260.
33. Liu, KY,Hu, S,Chan, BC(2013).Anti-inflammatory and anti-allergic activities of Pentaherb formula, Moutan Cortex (Danpi) and gallic acid.Molecules,18,2483-2500.
34. Liu, XW,Bernardo, MM,Fridman, R,Kim, HR(2003).Tissue inhibitor of metalloproteinase-1 protects human breast epithelial cells against intrinsic apoptotic cell death via the focal adhesion kinase/phosphatidylinositol 3-kinase and MAPK signaling pathway.J Biol Chem,278,40364-40372.
35. Lu, Z,Nie, G,Belton, PS,Tang, H,Zhao, B(2006).Structure-activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives.Neurochem Int,48,263-274.
36. Lukivskaya, O,Patsenker, E,Lis, R,Buko, VU(2008).Inhibition of inducible nitric oxide synthase activity prevents liver recovery in rat thioacetamide-induced fibrosis reversal.Eur J Clin Invest.,38,317-325.
37. Madrigal-Santillán, E,Madrigal-Bujaidar, E,Álvarez-González, I(2014).Review of natural products with hepatoprotective effects.World J Gastroenterol,20,14787-14804.
38. Maurya, DK,Devasagayam, TP(2010).Antioxidant and prooxidant nature of hydroxycinnamic acid derivatives ferulic and caffeic acids.Food Chem Toxicol,48,3369-3373.
39. Mayer, KE,Myers, RP,Lee, SS(2005).Silymarin treatment of viral hepatitis: a systematic review.J Viral Hepat,12,559-567.
40. Mosmann, T(1983).Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.J Immunol Meth,65,55-63.
41. Muriel, P,Rivera-Espinoza, Y.(2008).Beneficial drugs for liver diseases.J Appl Toxicol,28,93-103.
42. Ohkawa, H,Ohishi, N,Yagi, K(1979).Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction.Anal Biochem,95,351-358.
43. Pradhan, SC,Girish, C(2006).Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine.Indian J Med Res.,124,491-504.
44. Preaux, AM,D'ortho, MP,Bralet, MP,Laperche, Y,Mavier, P(2002).Apoptosis of human hepatic myofibroblasts promotes activation of matrix metalloproteinase-2.Hepatology,36,615-622.
45. Ramachandran, P,Iredale, JP(2012).Liver fibrosis: a bidirectional model of fibrogenesis and resolution.QJM,105,813-817.
46. Rao, HY,Wei, L,Li, J(2009).Liver fibrosis and hepatic stellate cells improvement of chronic hepatitis C patients by interferon-beta-1a with or without sustained viral response.Hepatogastroenterology,56,328-334.
47. Reitman, S,Frankel, S(1957).A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases.Am J Clin Pathol,28,56-63.
48. Seglen, PO(1976).Preparation of isolated rat liver cells.Meth Cell Biol,13,29-83.
49. Shen, HW,Yi, L,Wang, XM(2004).Expression of Caspase-3 and Bcl-2 in bladder transitional carcinoma and their significance.Ai Zheng,23,181-184.
50. Singh, RP,Chidambara Murthy, KN,Jayaprakasha, GK(2002).Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models.J Agric Food Chem,50,81-86.
51. Skehan, P,Storeng, R,Scudiero, D(1990).New colorimetric cytotoxicity assay for anticancer-drug screening.J Natl Cancer Inst,82,1107-1112.
52. Srivastava, A,Shivanandappa, T(2010).Stereospecificity in the cytotoxic action of hexachlorocyclohexane isomers.Chem Biol Interact,183,34-39.
53. Stupans, L,Tan, HW,Kirlich, A(2002).Inhibition of CYP3A-mediated oxidation in human hepatic microsomes by the dietary derived complex phenol, gallic acid.J Pharm Pharmacol,54,269-275.
54. Traister, A,Breitman, I,Bar-Lev, E(2005).Nicotinamide induces apoptosis and reduces collagen I and pro-inflammatory cytokines expression in rat hepatic stellate cells.Scand J Gastroenterol,40,1226-1234.
55. Troeger, JS,Mederacke, I,Gwak, GY(2012).Deactivation of hepatic stellate cells during liver fibrosis resolution in mice.Gastroenterology,143,1073-1083.
56. Tsukada, S,Parsons, CJ,Rippe, RA(2006).Mechanisms of liver fibrosis.Clin Chim Acta,364,33-60.
57. Upadhyaya, K,Hamidullah Singh, K,Arun, A(2016).Identification of gallic acid based glycoconjugates as a novel tubulin polymerization inhibitors.Org Biomol Chem,14,1338-1358.
58. Vijaya Padma, V,Sowmya, P,Arun Felix, T,Baskaran, R,Poornima, P(2011).Protective effect of gallic acid against lindane induced toxicity in experimental rats.Food Chem Toxicol,49,991-998.
59. Vogel, S,Piantedosi, R,Frank, J(2000).An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro.J Lipid Res,41,882-893.
60. Wang, J,Tang, L,White, J,Fang, J(2014).Inhibitory effect of gallic acid on CCl4-mediated liver fibrosis in mice.Cell Biochem Biophys,69,21-26.
61. Woessner, JF, Jr(1961).The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid.Arch Biochem Biophys,93,440-447.
62. Wynn, TA(2007).Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases.J Clin Invest,117,524-529.
63. Xie, H,Hou, W,Yang, Y,Yu, Y,Wang, F,Mao, J(2015).Effects of Shenqi Neijin powder on activation and apoptosis of hepatic stellate cells in rats with hepatic fibrosis.Int J Clin Exp Med,8,2226-2232.

1. Wisuit Pradidarcheep,Vichununt Kerdput,Suriya Pongsawat,Nattpawit Kaewnoonual,Cheng Nilbu-nga,Arunporn Itharat(2021).Therapeutic efficacy of a Dioscorea membranacea extract in a rat model of hepatocellular carcinoma: Histopathological aspects.Journal of Traditional and Complementary Medicine,11(5),400-408.