Alpha mangostin Inhibits Hepatic Stellate Cells Activation Through TGF-β/Smad and Akt Signaling Pathways: An in vitro Study in LX2

Abstract

Background Alpha mangostin has been reported to have activity for the treatment of liver fibrosis in the rats. However, the mechanisms of action are poorly understood. This study was aimed to investigate the effect of alpha mangostin on hepatic stellate cells (HSC) activation and proliferation through TGF-β/Smad and Akt signaling pathways.

Methods Immortalized HSC, LX2 cells, were incubated with TGF-β with or without alpha mangostin (5 or 10 μM). Sorafenib 10 µM was used as positive control. LX2 viability was counted using trypan blue exclusion method. The effect of alpha mangostin on TGF-β concentrations, and the expressions of proliferation and fibrogenic markers were evaluated.

Results Alpha mangostin treatment resulted in a reduced proliferation of HSC, decreased Ki-67 and p-Akt expressions. These findings were followed with decreased concentrations of TGF-β in the medium of cells treated with alpha mangostin, decreased expressions of COL1A1, TIMP1, PAI1, α-SMA, and p-Smad3 as fibrogenic markers. These effects were shown to be dose-dependent.

Conclusions Alpha mangostin inhibits hepatic stellate cells proliferation and activation through TGF-β/Smad and Akt signaling pathways in dose dependent manner.

Key words

Alpha mangostin - hepatic stellate cells - TGF-β - Smad - Akt

^* equal contribution

• References

• 1 Iredale JP, Thompson A, Henderson NC. Extracelluler matrix degradation in liver fibrosis: Biochemistry and regulation. Biochim Biophys Acta 2013; 876-883
• 2 Mallat A. Lotersztajn Cellular Mechanisms of Tissue Fibrosis. 5. Novel insights into liver fibrosis. J Physiol Cell Physiol 2013; 305: 789-799
• 3 Bataller R, Brenner DA. Liver fibrosis. J Clin Invest 2005; 115: 209-218 Review. Erratum in: J Clin Invest. 2005;115(4):1100
• 4 Xu F, Liu C, Zhou D. et al. TGF-β/SMAD pathway and its regulation in hepatic fibrosis. J Histochem Cytochem. 2016; 64: 157-167
• 5 Gressner AM, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cell and TGF-β as major players and therapeutic targets. J Cell Mol Med. 2006; 10: 76-99
• 6 Fabregat I, Moreno-Càceres J, Sánchez A. et al. TGF-β Signaling and Liver Disease. FEBS J. 2016; 283: 2219-2232
• 7 Dooley S, Delvoux B, Lahme B. et al. Modulation of transforming growth factor beta response and signaling during transdifferentiation of rat hepatic stellate cells to myofibroblasts. Hepatology 2000; 31: 1094-1106
• 8 Kang Y, Kim B, Park YK. et al. Astaxanthin prevents TGFβ1-induced pro-fibrogenic gene expression by inhibiting Smad3 activation in hepatic stellate cells. Biochim Biophys Acta. 2015; 1850: 178-185
• 9 Mu Y, Gudey SK, Landstorm M. Non-smad signaling pathways. Cell Tissue Res. 2012; 347: 11-20
• 10 Son G, Hines IN, Lindquist J. et al. Inhibition of phosphatidylinositol 3-kinase signaling in hepatic stellate cells blocks the progression of hepatic fibrosis. J Hepatol. 2009; 50: 1512-1523
• 11 Ghosh AK, Vaughan DE. PAI-1 in Tissue Fibrosis. J Cell Physiol. 2012; 227: 493-507
• 12 Nakerakanti S, Trojanowska M. The role of TGF-β receptors in fibrosis. Rheumatol J. 2012; 6: 156-162
• 13 Ibrahim MY, Hashim NM, Mariod AA. et al. α-Mangostin from Garcinia mangostana Linn: An updated review of its pharmacological properties. Arabian J Chem 2016; 9: 317-329
• 14 Zhu D, He X, Duan Y. et al. Expression of microRNA-454 in TGF-β1-stimulated hepatic stellate cells and in mouse livers infected with Schistosoma japonicum. Parasit Vectors 2014; 7: 148
• 15 Kang KH, Qian ZJ, Ryu B. et al. Hepatic fibrosis inhibitory effect of peptides isolated from navicula incerta on TGF-β1 induced activation of lx-2 human hepatic stellate cells. Prev Nutr Food Sci 2013; 18: 124-132
• 16 Zhang ZY, Chu RY, Zhou XT. et al. The expression of Ki 67, TGF-β2, and α-SMA after the femtosecond laser assisted intrastromal scanning in rabit cornea. Zhonghua Yan Ke Za Zhi 2010; 46: 214-220
• 17 Hall MC, Young DA, Waters JG. et al. The comparative role of activator protein 1 and smad factors in the regulation of timp-1 and mmp-1 gene expression by transforming growth factor-β1. J Biol Chem. 2003; 278: 10304-10313
• 18 Tang LX, He RH, Yang G. et al. Asiatic acid inhibits liver fibrosis by blocking TGF-beta/Smad signaling in vivo and in vitro. PLoS One 2012; 7: e31350
• 19 Shi YF, Zhang Q, Cheung PY. et al. Effects of rhDecorin on TGF-beta1 induced human hepatic stellate cells LX-2 activation. Biochim Biophys Acta. 2006; 1760: 1587-1595
• 20 Dumont N, Bakin AV, Arteaga CL. Autocrine Transforming Growth Factor-β signaling mediates Smad independent motility in human cancer cells. J Biol Chem. 2003; 278: 3275-3285
• 21 Koyama Y, Brenner DA. New therapies for hepatic fibrosis. Clin Res Hepatol Gastroenterol 2015; 39 (Suppl. 01) S75-S79
• 22 Fagone P, Mangano K, Pesce A. et al. Emerging therapeutic targets for the treatment of hepatic fibrosis. Drug Discov Today. 2016; 21: 369-375
• 23 Hong F, Chou H, Fiel I. et al. Antifibrotic activity of sorafenib in experimental hepatic fibrosis-refinement of inhibitory targets, dosing and window of efficacy in vivo. Dig Dis Sci. 2013; 58: 257-264
• 24 Zhang CZ, Wang XD, Wang HW. et al. Sorafenib inhibits liver cancer growth by decreasing mTOR, AKT, and PI3K expression. J Buon. 2015; 20: 218-222
• 25 Stefano JT, Pereira IVA, Torres MM. et al. Sorafenib prevents liver fibrosis in a non-alcoholic steatohepatitis (NASH) rodent model. Braz J Med Biol Res. 2015; 48: 408-414
• 26 Toricelli M, Melo FHM, Hunger A. et al. Timp1 promotes cell survival by acting the PDK1 signaling pathway in melanoma. Cancers 2017; 9: 1-15