MEK inhibition prevents TGF‑β1‑induced myofibroblast transdifferentiation in human tenon fibroblasts

Wen,Jiamin; Lin,Xianchai; Gao,Wuyou; Qu,Bo; Ling,Yunlan; Liu,Rongjiao; Yu,Minbin

doi:10.3892/mmr.2018.9673

MEK inhibition prevents TGF‑β1‑induced myofibroblast transdifferentiation in human tenon fibroblasts

Authors:
- Jiamin Wen
- Xianchai Lin
- Wuyou Gao
- Bo Qu
- Yunlan Ling
- Rongjiao Liu
- Minbin Yu
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Affiliations: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat‑sen University, Guangzhou, Guangdong 510060, P.R. China
Published online on: November 20, 2018 https://doi.org/10.3892/mmr.2018.9673
Pages: 468-476
Copyright: © Wen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Subconjunctival fibrosis represents the primary cause of postoperative failure of trabeculectomy, and at present there is a lack of effective intervention strategies. The present study aimed to investigate the effect of the mitogen‑activated protein kinase kinase (MEK) inhibitor U0126 on human tenon fibroblast (HTF) myofibrosis transdifferentiation, and to illuminate the underlying molecular mechanisms involved. It was demonstrated that U0126 significantly inhibited the proliferation, migration and collagen contraction of HTFs stimulated with TGF‑β1. In addition, U0126 largely attenuated the TGF‑β1‑induced conversion of HTFs into myofibroblasts, as indicated by a downregulation of the mRNA and protein expression of α‑smooth muscle actin and zinc finger protein SNAI1, and by ameliorating the 3D‑collagen contraction response. Mechanistically, U0126 suppressed the TGF‑β1‑stimulated phosphorylation of mothers against decapentaplegic homolog 2/3, P38 mitogen‑activated protein kinase and extracellular signal‑regulated kinase 1/2, indicating that U0126 may inhibit HTF activation through the canonical and non‑canonical signaling pathways of TGF‑β1. Therefore, U0126 exhibits a potent anti‑fibrotic effect among HTFs, and the inhibition of MEK signaling may serve as an alternative intervention strategy for the treatment of trabeculectomy‑associated fibrosis.

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1	Fan Gaskin JC, Nguyen DQ, Soon Ang G, O'Connor J and Crowston JG: Wound healing modulation in glaucoma filtration surgery-conventional practices and new perspectives: The role of antifibrotic agents (Part I). J Curr Glaucoma Pract. 8:37–45. 2014. View Article : Google Scholar : PubMed/NCBI
2	Pimentel E and Schmidt J: Is mytomicyn better than 5-fluorouracil as antimetabolite in trabeculectomy for glaucoma? Medwave. 18:e71372018.(In English, Spanish). View Article : Google Scholar : PubMed/NCBI
3	Kitazawa Y, Kawase K, Matsushita H and Minobe M: Trabeculectomy with mitomycin. A comparative study with fluorouracil. Arch Ophthalmol. 109:1693–1698. 1991. View Article : Google Scholar : PubMed/NCBI
4	Shi H, Wang H, Fu S, Xu K, Zhang X, Xiao Y and Ye W: Losartan attenuates scar formation in filtering bleb after trabeculectomy. Invest Ophthalmol Vis Sci. 58:1478–1486. 2017. View Article : Google Scholar : PubMed/NCBI
5	Chang L, Crowston JG, Cordeiro MF, Akbar AN and Khaw PT: The role of the immune system in conjunctival wound healing after glaucoma surgery. Surv Ophthalmol. 45:49–68. 2000. View Article : Google Scholar : PubMed/NCBI
6	Hinz B, Phan SH, Thannickal VJ, Prunotto M, Desmoulière A, Varga J, De Wever O, Mareel M and Gabbiani G: Recent developments in myofibroblast biology: Paradigms for connective tissue remodeling. Am J Pathol. 180:1340–1355. 2012. View Article : Google Scholar : PubMed/NCBI
7	Hartupee J and Mann DL: Role of inflammatory cells in fibroblast activation. J Mol Cell Cardiol. 93:143–148. 2016. View Article : Google Scholar : PubMed/NCBI
8	Carthy JM: TGFβ signaling and the control of myofibroblast differentiation: Implications for chronic inflammatory disorders. J Cell Physiol. 233:98–106. 2018. View Article : Google Scholar : PubMed/NCBI
9	Nickel J, Ten Dijke P and Mueller TD: TGF-β family co-receptor function and signaling. Acta Biochim Biophys Sin (Shanghai). 50:12–36. 2018. View Article : Google Scholar : PubMed/NCBI
10	Lin X, Yu M, Wu K, Yuan H and Zhong H: Effects of pirfenidone on proliferation, migration, and collagen contraction of human Tenon's fibroblasts in vitro. Invest Ophthalmol Vis Sci. 50:3763–3770. 2009. View Article : Google Scholar : PubMed/NCBI
11	Li DQ, Lee SB and Tseng SC: Differential expression and regulation of TGF-beta1, TGF-beta2, TGF-beta3, TGF-betaRI, TGF-betaRII and TGF-betaRIII in cultured human corneal, limbal, and conjunctival fibroblasts. Curr Eye Res. 19:154–161. 1999. View Article : Google Scholar : PubMed/NCBI
12	Fu S, Sun L, Zhang X, Shi H, Xu K, Xiao Y and Ye W: 5-Aza-2′-deoxycytidine induces human Tenon's capsule fibroblasts differentiation and fibrosis by up-regulating TGF-β type I receptor. Exp Eye Res. 165:47–58. 2017. View Article : Google Scholar : PubMed/NCBI
13	Stahnke T, Kowtharapu BS, Stachs O, Schmitz KP, Wurm J, Wree A, Guthoff RF and Hovakimyan M: Suppression of TGF-β pathway by pirfenidone decreases extracellular matrix deposition in ocular fibroblasts in vitro. PLoS One. 12:e01725922017. View Article : Google Scholar : PubMed/NCBI
14	Meyer-Ter-Vehn T, Gebhardt S, Sebald W, Buttmann M, Grehn F, Schlunck G and Knaus P: p38 inhibitors prevent TGF-beta-induced myofibroblast transdifferentiation in human tenon fibroblasts. Invest Ophthalmol Vis Sci. 47:1500–1509. 2006. View Article : Google Scholar : PubMed/NCBI
15	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
16	Shu DY and Lovicu FJ: Myofibroblast transdifferentiation: The dark force in ocular wound healing and fibrosis. Prog Retin Eye Res. 60:44–65. 2017. View Article : Google Scholar : PubMed/NCBI
17	Hsu CK, Lin HH, Harn HI, Hughes MW, Tang MJ and Yang CC: Mechanical forces in skin disorders. J Dermatol Sci. 90:232–240. 2018. View Article : Google Scholar : PubMed/NCBI
18	Liu Y, Kimura K, Orita T, Teranishi S, Suzuki K and Sonoda KH: Inhibition by all-trans-retinoic acid of transforming growth factor-β-induced collagen gel contraction mediated by human tenon fibroblasts. Invest Ophthalmol Vis Sci. 55:4199–4205. 2014. View Article : Google Scholar : PubMed/NCBI
19	Meyer-ter-Vehn T, Sieprath S, Katzenberger B, Gebhardt S, Grehn F and Schlunck G: Contractility as a prerequisite for TGF-beta-induced myofibroblast transdifferentiation in human tenon fibroblasts. Invest Ophthalmol Vis Sci. 47:4895–4904. 2006. View Article : Google Scholar : PubMed/NCBI
20	Wallace HA and Bhimji SS: Wound, Healing, Phases, in StatPearls. StatPearls Publishing StatPearls Publishing LLC; Treasure Island (FL): 2017
21	Reiter C, Wimmer S, Schultheiss A, Klink T, Grehn F and Geerling G: Corneal epitheliopathy following trabeculectomy with postoperative adjunctive 5-fluorouracil. Klin Monbl Augenheilkd. 227:887–891. 2010.(In German). View Article : Google Scholar : PubMed/NCBI
22	Fan Gaskin JC, Nguyen DQ, Soon Ang G, O'Connor J and Crowston JG: Wound healing modulation in glaucoma filtration surgery-conventional practices and new perspectives: Antivascular endothelial growth factor and novel agents (Part II). J Curr Glaucoma Pract. 8:46–53. 2014. View Article : Google Scholar : PubMed/NCBI
23	Bindlish R, Condon GP, Schlosser JD, D'Antonio J, Lauer KB and Lehrer R: Efficacy and safety of mitomycin-C in primary trabeculectomy: Five-year follow-up. Ophthalmology. 109:1336–1342. 2002. View Article : Google Scholar : PubMed/NCBI
24	Khaw PT, Sherwood MB, Doyle JW, Smith MF, Grierson I, McGorray S and Schultz GS: Intraoperative and post operative treatment with 5-fluorouracil and mitomycin-c: Long term effects in vivo on subconjunctival and scleral fibroblasts. Int Ophthalmol. 16:381–385. 1992. View Article : Google Scholar : PubMed/NCBI
25	Chang HH, Chang MC, Wu IH, Huang GF, Huang WL, Wang YL, Lee SY, Yeh CY, Guo MK, Chan CP, et al: Role of ALK5/Smad2/3 and MEK1/ERK signaling in transforming growth factor beta 1-modulated growth, collagen turnover and differentiation of stem cells from apical papilla of human tooth. J Endod. 41:1272–1280. 2015. View Article : Google Scholar : PubMed/NCBI
26	Chen X, Ye S, Xiao W, Wang W, Luo L and Liu Y: ERK1/2 pathway mediates epithelial-mesenchymal transition by cross-interacting with TGFβ/Smad and Jagged/Notch signaling pathways in lens epithelial cells. Int J Mol Med. 33:1664–1670. 2014. View Article : Google Scholar : PubMed/NCBI
27	Nickells RW: The cell and molecular biology of glaucoma: mechanisms of retinal ganglion cell death. Invest Ophthalmol Vis Sci. 53:2476–2481. 2012. View Article : Google Scholar : PubMed/NCBI
28	Gupta VK, You Y, Li JC, Klistorner A and Graham SL: Protective effects of 7,8-dihydroxyflavone on retinal ganglion and RGC-5 cells against excitotoxic and oxidative stress. J Mol Neurosci. 49:96–104. 2013. View Article : Google Scholar : PubMed/NCBI
29	Wei J, Jiang H, Gao H and Wang G: Raf-1 kinase inhibitory protein (RKIP) promotes retinal ganglion cell survival and axonal regeneration following optic nerve crush. J Mol Neurosci. 57:243–248. 2015. View Article : Google Scholar : PubMed/NCBI
30	Luo JM, Cen LP, Zhang XM, Chiang SW, Huang Y, Lin D, Fan YM, van Rooijen N, Lam DS, Pang CP and Cui Q: PI3K/akt, JAK/STAT and MEK/ERK pathway inhibition protects retinal ganglion cells via different mechanisms after optic nerve injury. Eur J Neurosci. 26:28–42. 2007. View Article : Google Scholar
31	Hinz B, Mastrangelo D, Iselin CE, Chaponnier C and Gabbiani G: Mechanical tension controls granulation tissue contractile activity and myofibroblast differentiation. Am J Pathol. 159:1009–1020. 2001. View Article : Google Scholar : PubMed/NCBI
32	Biswas H and Longmore GD: Action of SNAIL1 in cardiac myofibroblasts is important for cardiac fibrosis following hypoxic injury. PLoS One. 11:e01626362016. View Article : Google Scholar : PubMed/NCBI
33	Reichard JF and Petersen DR: Involvement of phosphatidylinositol 3-kinase and extracellular-regulated kinase in hepatic stellate cell antioxidant response and myofibroblastic transdifferentiation. Arch Biochem Biophys. 446:111–118. 2006. View Article : Google Scholar : PubMed/NCBI
34	Makino T, Jinnin M, Muchemwa FC, Fukushima S, Kogushi-Nishi H, Moriya C, Igata T, Fujisawa A, Johno T and Ihn H: Basic fibroblast growth factor stimulates the proliferation of human dermal fibroblasts via the ERK1/2 and JNK pathways. Br J Dermatol. 162:717–723. 2010. View Article : Google Scholar : PubMed/NCBI
35	Derynck R and Zhang YE: Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 425:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
36	Guo X and Wang XF: Signaling cross-talk between TGF-beta/BMP and other pathways. Cell Res. 19:71–88. 2009. View Article : Google Scholar : PubMed/NCBI
37	Pardali E, Sanchez-Duffhues G, Gomez-Puerto MC and Ten Dijke P: TGF-β-induced endothelial-mesenchymal transition in fibrotic diseases. Int J Mol Sci. 18(pii): E21572017. View Article : Google Scholar : PubMed/NCBI
38	Zeglinski MR, Roche P, Hnatowich M, Jassal DS, Wigle JT, Czubryt MP and Dixon IM: TGFβ1 regulates Scleraxis expression in primary cardiac myofibroblasts by a Smad-independent mechanism. Am J Physiol Heart Circ Physiol. 310:H239–H249. 2016. View Article : Google Scholar : PubMed/NCBI
39	Che X, Wang Q, Xie Y, Xu W, Shao X, Mou S and Ni Z: Astragaloside IV suppresses transforming growth factor-β1 induced fibrosis of cultured mouse renal fibroblasts via inhibition of the MAPK and NF-κB signaling pathways. Biochem Biophys Res Commun. 464:1260–1266. 2015. View Article : Google Scholar : PubMed/NCBI
40	Kamaraju AK and Roberts AB: Role of Rho/ROCK and p38 MAP kinase pathways in transforming growth factor-beta-mediated Smad-dependent growth inhibition of human breast carcinoma cells in vivo. J Biol Chem. 280:1024–1036. 2005. View Article : Google Scholar : PubMed/NCBI
41	Iwayama H, Sakamoto T, Nawa A and Ueda N: Crosstalk between Smad and mitogen-activated protein kinases for the regulation of apoptosis in cyclosporine A- induced renal tubular injury. Nephron Extra. 1:178–189. 2011. View Article : Google Scholar : PubMed/NCBI
42	Zhu Y, Gu J, Zhu T, Jin C, Hu X and Wang X: Crosstalk between Smad2/3 and specific isoforms of ERK in TGF-β1-induced TIMP-3 expression in rat chondrocytes. J Cell Mol Med. 21:1781–1790. 2017. View Article : Google Scholar : PubMed/NCBI
43	Boye A, Kan H, Wu C, Jiang Y, Yang X, He S and Yang Y: MAPK inhibitors differently modulate TGF-β/Smad signaling in HepG2 cells. Tumour Biol. 36:3643–3651. 2015. View Article : Google Scholar : PubMed/NCBI