Inhibitory effect of tranilast on the myofibroblast differentiation of rat mesenchymal stem cells induced by transforming growth factor‑β1 in vitro

Tang,Wenxian; Zhang,Yuejuan; Tang,Lin; Zhang,Jun; Xiong,Lei; Wang,Baohe

doi:10.3892/mmr.2018.9588

Inhibitory effect of tranilast on the myofibroblast differentiation of rat mesenchymal stem cells induced by transforming growth factor‑β1 in vitro

Authors:
- Wenxian Tang
- Yuejuan Zhang
- Lin Tang
- Jun Zhang
- Lei Xiong
- Baohe Wang
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Affiliations: College of Life Science, Hunan Normal University, Changsha, Hunan 410081, P.R. China, Department of Biochemistry and Molecular Biology, College of Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, P.R. China
Published online on: October 24, 2018 https://doi.org/10.3892/mmr.2018.9588
Pages: 5693-5700

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Abstract

Mesenchymal stem cell (MSC) transplantation is able to attenuate organ fibrosis; however, increasing evidence has indicated that MSCs may be an important cell source of myofibroblasts, which are vital pathogenic cells in fibrotic diseases. The results of the present study revealed that co‑culturing with exogenous transforming growth factor (TGF)‑β1 can induce the transdifferentiation of cultured rat MSCs into myofibroblasts in vitro. Treatment of the MSCs with tranilast [N‑(3',4'‑dimethoxycinnamoyl)‑anthranilic acid] attenuated this fibrotic process. Immunocytochemical staining, western blot analysis, reverse transcription‑quantitative polymerase chain reaction analysis and cell viability assays were performed in order to evaluate the molecular mechanisms underlying the effects of tranilast on TGF‑β1‑mediated MSC‑to‑myofibroblast activation. The results demonstrated that TGF‑β1 upregulated the expression of α‑smooth muscle actin (α‑SMA) and collagen type I, and increased the phosphorylation of mothers against decapentaplegic homolog 3 (Smad3) and extracellular signal‑regulated kinase 1/2 (ERK1/2) in the rat MSCs; by contrast, tranilast pretreatment downregulated their expression. Furthermore, the proliferation of MSCs induced by TGF‑β1 was decreased by pretreatment with tranilast. In conclusion, the results of the present study demonstrated that tranilast treatment markedly suppressed the TGF‑β1‑induced differentiation of cultured rat MSCs into myofibroblasts, potentially by inhibiting the Smad3 and ERK1/2 signaling pathways. Therefore, this may be a potential antifibrotic therapeutic strategy, serving as an adjuvant treatment following transplantation of MSCs.

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1	Wynn TA: Cellular and molecular mechanisms of fibrosis. J Pathol. 214:199–210. 2008. View Article : Google Scholar : PubMed/NCBI
2	El Agha E, Kramann R, Schneider RK, Li X, Seeger W, Humphreys BD and Bellusci S: Mesenchymal stem cells in fibrotic disease. Cell Stem Cell. 21:166–177. 2017. View Article : Google Scholar : PubMed/NCBI
3	Hinz B: Formation and function of the myofibroblast during tissue repair. J Invest Dermatol. 127:526–537. 2007. View Article : Google Scholar : PubMed/NCBI
4	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003. View Article : Google Scholar : PubMed/NCBI
5	LeBleu VS, Taduri G, O'Connell J, Teng Y, Cooke VG, Woda C, Sugimoto H and Kalluri R: Origin and function of myofibroblasts in kidney fibrosis. Nat Med. 19:1047–1053. 2013. View Article : Google Scholar : PubMed/NCBI
6	Kramann R, Schneider RK, DiRocco DP, Machado F, Fleig S, Bondzie PA, Henderson JM, Ebert BL and Humphreys BD: Perivascular Gli1+ progenitors are key contributors to injury-induced organ fibrosis. Cell Stem Cell. 16:51–66. 2015. View Article : Google Scholar : PubMed/NCBI
7	Bonfield TL and Caplan AI: Adult mesenchymal stem cells: An innovative therapeutic for lung diseases. Discov Med. 9:337–345. 2010.PubMed/NCBI
8	Baertschiger RM, Serre-Beinier V, Morel P, Bosco D, Peyrou M, Clément S, Sgroi A, Kaelin A, Buhler LH and Gonelle-Gispert C: Correction: Fibrogenic potential of human multipotent mesenchymal stromal cells in injured liver. PLoS One. 4:e66572009. View Article : Google Scholar : PubMed/NCBI
9	Russo FP, Alison MR, Bigger BW, Amofah E, Florou A, Amin F, Bou-Gharios G, Jeffery R, Iredale JP and Forbes SJ: The bone marrow functionally contributes to liver fibrosis. Gastroenterology. 130:1807–1821. 2006. View Article : Google Scholar : PubMed/NCBI
10	Tang N, Zhao Y, Feng R, Liu Y, Wang S, Wei W, Ding Q, An MS, Wen J and Li L: Lysophosphatidic acid accelerates lung fibrosis by inducing differentiation of mesenchymal stem cells into myofibroblasts. J Cell Mol Med. 18:156–169. 2014. View Article : Google Scholar : PubMed/NCBI
11	Carlson S, Trial J, Soeller C and Entman ML: Cardiac mesenchymal stem cells contribute to scar formation after myocardial infarction. Cardiovasc Res. 91:99–107. 2011. View Article : Google Scholar : PubMed/NCBI
12	Yang S, Cui H, Xie N, Icyuz M, Banerjee S, Antony VB, Abraham E, Thannickal VJ and Liu G: miR-145 regulates myofibroblast differentiation and lung fibrosis. FASEB J. 27:2382–2391. 2013. View Article : Google Scholar : PubMed/NCBI
13	Bianco P, Cao X, Frenette PS, Mao JJ, Robey PG, Simmons PJ and Wang CY: The meaning, the sense and the significance: Translating the science of mesenchymal stem cells into medicine. Nat Med. 19:35–42. 2013. View Article : Google Scholar : PubMed/NCBI
14	Xu J, Lamouille S and Derynck R: TGF-beta-induced epithelial to mesenchymal transition. Cell Res. 19:156–172. 2009. View Article : Google Scholar : PubMed/NCBI
15	Wang W, Koka V and Lan HY: Transforming growth factor-beta and Smad signalling in kidney diseases. Nephrology (Carlton). 10:48–56. 2005. View Article : Google Scholar : PubMed/NCBI
16	Peng H, Carretero OA, Peterson EL and Rhaleb NE: Ac-SDKP inhibits transforming growth factor-beta1-induced differentiation of human cardiac fibroblasts into myofibroblasts. Am J Physiol Heart Circ Physiol. 298:H1357–H1364. 2010. View Article : Google Scholar : PubMed/NCBI
17	Sun Q, Wu Y, Zhao F and Wang J: Maresin 1 inhibits transforming growth factor-β1-induced proliferation, migration and differentiation in human lung fibroblasts. Mol Med Rep. 16:1523–1529. 2017. View Article : Google Scholar : PubMed/NCBI
18	Darakhshan S and Pour AB: Tranilast: A review of its therapeutic applications. Pharmacol Res. 91:15–28. 2015. View Article : Google Scholar : PubMed/NCBI
19	Yamada H, Tajima S, Nishikawa T, Murad S and Pinnell SR: Tranilast, a selective inhibitor of collagen synthesis in human skin fibroblasts. J Biochem. 116:892–897. 1994. View Article : Google Scholar : PubMed/NCBI
20	Isaji M, Nakajoh M and Naito J: Selective inhibition of collagen accumulation by N-(3,4-dimethoxycinnamoyl)anthranilic acid (N-5′) in granulation tissue. Biochem Pharmacol. 36:469–474. 1987. View Article : Google Scholar : PubMed/NCBI
21	Kim TI, Lee H, Hong HK, Kim KS, Choi SI, Maeng YS and Kim EK: Inhibitory effect of tranilast on transforming growth factor-beta-induced protein in granular corneal dystrophy type 2 corneal fibroblasts. Cornea. 34:950–958. 2015. View Article : Google Scholar : PubMed/NCBI
22	Nakatani Y, Nishida K, Sakabe M, Kataoka N, Sakamoto T, Yamaguchi Y, Iwamoto J, Mizumaki K, Fujiki A and Inoue H: Tranilast prevents atrial remodeling and development of atrial fibrillation in a canine model of atrial tachycardia and left ventricular dysfunction. J Am Coll Cardiol. 61:582–588. 2013. View Article : Google Scholar : PubMed/NCBI
23	See F, Watanabe M, Kompa AR, Bing HW, Boyle AJ, Kelly DJ, Gilbert RE and Krum H: Early and delayed tranilast treatment reduces pathological fibrosis following myocardial infarction. Heart Lung Circ. 22:122–132. 2013. View Article : Google Scholar : PubMed/NCBI
24	Popova AP, Bozyk PD, Goldsmith AM, Linn MJ, Lei J, Bentley JK and Hershenson MB: Autocrine production of TGF-beta1 promotes myofibroblastic differentiation of neonatal lung mesenchymal stem cells. Am J Physiol Lung Cell Mol Physiol. 298:L735–L743. 2010. View Article : Google Scholar : PubMed/NCBI
25	Kakudo N, Kushida S, Suzuki K, Ogura T, Notodihardjo PV, Hara T and Kusumoto K: Effects of transforming growth factor-beta1 on cell motility, collagen gel contraction, myofibroblastic differentiation, and extracellular matrix expression of human adipose-derived stem cell. Human Cell. 25:87–95. 2012. View Article : Google Scholar : PubMed/NCBI
26	Song K, Huang M, Shi Q, Du T and Cao Y: Cultivation and identification of rat bone marrow-derived mesenchymal stem cells. Mol Med Rep. 10:755–760. 2014. View Article : Google Scholar : PubMed/NCBI
27	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
28	Phan SH: The myofibroblast in pulmonary fibrosis. Chest. 122 6 Suppl:286S–289S. 2002. View Article : Google Scholar : PubMed/NCBI
29	di Bonzo LV, Ferrero I, Cravanzola C, Mareschi K, Rustichell D, Novo E, Sanavio F, Cannito S, Zamara E, Bertero M, et al: Human mesenchymal stem cells as a two-edged sword in hepatic regenerative medicine: Engraftment and hepatocyte differentiation versus profibrogenic potential. Gut. 57:223–231. 2008. View Article : Google Scholar : PubMed/NCBI
30	Tang N, Zhao Y, Feng R, Liu Y, Wang S, Wei W, Ding Q, An MS, Wen J and Li L: Lysophosphatidic acid accelerates lung fibrosis by inducing differentiation of mesenchymal stem cells into myofibroblasts. J Cell Mol Med. 18:156–169. 2014. View Article : Google Scholar : PubMed/NCBI
31	Yasukawa T, Kimura H, Dong J, Tabata Y, Miyamoto H, Honda Y and Ogura Y: Effect of tranilast on proliferation, collagen gel contraction, and transforming growth factor beta secretion of retinal pigment epithelial cells and fibroblasts. Ophthalmic Res. 34:206–212. 2002. View Article : Google Scholar : PubMed/NCBI
32	Hattori T and Wang PL: Inhibition by tranilast of nifedipine-induced proliferation of cultured human gingival fibroblasts. Eur J Pharmacol. 498:79–81. 2004. View Article : Google Scholar : PubMed/NCBI
33	Wang S, Meng XM, Ng YY, Ma FY, Zhou S, Zhang Y, Yang C, Huang XR, Xiao J, Wang YY, et al: TGF-β/Smad3 signalling regulates the transition of bone marrow-derived macrophages into myofibroblasts during tissue fibrosis. Oncotarget. 7:8809–8822. 2016.PubMed/NCBI
34	Bonniaud P, Kolb M, Galt T, Robertson J, Robbins C, Stampfli M, Lavery C, Margetts PJ, Roberts AB and Gauldie J: Smad3 null mice develop airspace enlargement and are resistant to TGF-beta-mediated pulmonary fibrosis. J Immunol. 173:2099–2108. 2004. View Article : Google Scholar : PubMed/NCBI
35	Sato M, Muragaki Y, Saika S, Roberts AB and Ooshima A: Targeted disruption of TGF-β1/Smad3 signaling protects against renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction. J Clin Invest. 112:1486–1494. 2003. View Article : Google Scholar : PubMed/NCBI
36	Gu L, Zhu YJ, Yang X, Guo ZJ, Xu WB and Tian XL: Effect of TGF-beta/Smad signaling pathway on lung myofibroblast differentiation. Acta Pharmacol Sin. 28:382–391. 2007. View Article : Google Scholar : PubMed/NCBI