ISSN - 0975-7058 Vol 11, Special Issue 1, 2019 ISSN - 0975-7058 THE EFFECT OF ALPHA-MANGOSTIN ON TRANSFORMING GROWTH FACTOR BETA 1 (TGF-β1) AND MATRIX METALLOPROTEINASE-3 EXPRESSION IN TGF-β-INDUCED HEPATIC STELLATE CELLS SYARINTA ADENINA 1 , RAHMANIAH RAHMANIAH 1 , YUYUNTIA YUYUNTIA 1 , VIVIAN SOETIKNO 2 , MELVA LOUISA 2 * 1 Master Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia. 2 Department of Pharmacology and Therapeutics, Faculty of Medicine, Universitas Indonesia. Email: melva.louisa@gmail.com Received: 20 August 2018, Revised and Accepted: 07 February 2019 ABSTRACT Objective: Alpha-mangostin (α-MG) has been shown to possess antifibrotic effects. However, the specific mechanism of action of this compound remains poorly understood. Therefore, the aim of this study was to investigate the effect of α-MG on the expression levels of transforming growth factor (TGF)-β1 and matrix metalloproteinase-3 (MMP3) in hepatic stellate cells (HSCs) induced by TGF-β. Methods: Immortalized HSCs and LX-2 cells were incubated with TGF-β with or without α-MG (5 and 10 μM). The viability of LX-2 cells was assessed using the Trypan Blue Exclusion Method. The effect of α-MG on cell morphology and the mRNA expression levels of TGF-β1, TGF-β receptor, and MMP3 was then evaluated. Results: TGF-β enhanced the proliferation of HSCs and caused significant increases in the expression levels of TGF-β1, TGF-β receptor, and MMP3. α-MG treatment reduced the proliferation of HSCs and decreased the expression levels of TGF-β1, TGF-β1 receptor, and MMP3. Conclusion: α-MG is a potential antifibrotic agent due to its antiproliferative and antifibrogenic effects, mainly by suppressing the expression of TGF-β and MMP3 on the surfaces of activated HSCs. Keywords: Alpha-mangostin, Transforming growth factor-β, Matrix metalloproteinase-3, Hepatic stellate cells. INTRODUCTION Hepatic stellate cells (HSCs) are liver-specific mesenchymal cells that play a role in the pathogenesis of liver fibrosis [1,2]. HSCs are located in the perisinusoidal space and contain lipid droplets rich in Vitamin A. In the normal liver, stellate cells remain quiescent and play a role in maintaining the basal balance of the matrix membrane [2,3]. In response to liver injury, these cells receive signals from a wide variety of growth factors, cytokines, lipid mediators, and adipokines to promote survival at the sites of liver injury containing damaged hepatocytes and immune cells. HSCs then transdifferentiate to activated myofibroblast- like cells [2]. Transforming growth factor-β (TGF-β) is a key profibrogenic mediator that plays a major role in the activation of HSCs. Activated HSCs then secrete TGF-β in autocrine- and paracrine-dependent manners to mediate the production, degradation, and accumulation of molecules in the extracellular matrix (ECM) [1,4–6]. In humans, TGF-β exists in three homologous isoforms: TGF-β1, TGF-β2, and TGF-β3. On binding to TGF-β receptor type II, TGF-β consecutively recruits TGF-β receptor type I to the complex and initiates TGF-β signaling, which causes the activation of SMAD-dependent and SMAD-independent pathways [7,8]. Activated HSCs express many ECM-associated proteins, including collagen type I, α-smooth muscle actin, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs) [4,9]. Accumulation of these proteins by the ECM without sufficient degradation will form deposits that destroy and distort the normal hepatic structure, which results in cirrhosis and liver failure [1,2,6,8,10]. ECM degradation is mediated by MMPs, a family of zinc-dependent enzymes grouped into collagenases, gelatinases, stromelysins, and membrane-type MMPs. TIMPs regulate the MMP activity by binding to MMPs, thereby inhibiting proteolytic activities [11]. Although the TGF-β pathway is important in the pathogenesis of liver fibrosis, few targeted drugs have been developed. Pirfenidone, a pyridine derivate, is the only drug currently available to specifically block TGF-β1 production for the long-term improvement of liver inflammation and fibrosis [9]. In addition, sorafenib is the only drug approved for the treatment of advanced-stage hepatocellular carcinoma. Hence, the development of new drugs and therapeutic regimens is an immediate medical necessity [12]. Various herbal products have been shown anti-liver fibrogenic activities by blocking TGF-β signaling and HSC activation [13]. For example, alpha-mangostin (α-MG), a xanthone derivate, is a major bioactive compound found in the tropical fruit Garcinia mangostana Linn. that has antifibrotic and antiproliferative activities, and has been shown to decrease TGF-β levels. However, the mechanism of action of this compound remains poorly understood. Therefore, the aim of the present study was to evaluate the effect of α-MG on the expression of TGF-β1 and its receptors, and MMP-3 in HSCs induced by TGF-β [3,14]. METHODS Cell culture Immortalized human HSCs and LX-2 cells were cultured as described elsewhere [3]. Cell treatments Cells were treated with one of four regimens in 10-cm culture dishes: (1) Medium only; (2) TGF-β at 2 ng/mL for 24 h, then new medium with TGF-β at 2 ng/mL for 24 h; (3) TGF-β at 2 ng/mL for 24 h, then new medium with TGF-β at 2 ng/mL and α-MG at 5 µM for 24 h; or (4) TGF-β at 2 ng/mL for 24 h, then new medium with TGF-β at 2 ng/mL and α-MG at 10 µM for 24 h. Afterward, the cells were harvested using Research Article © 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ijap.2019.v11s1.16144 The 3 rd International Conference on Global Health (ICGH), Universitas Indonesia, Bali, Indonesia