Celastrol protects TGF-β1-induced endothelial-mesenchymal transition

Abstract

The endothelial-to-mesenchymal transition (EndMT) in endothelial cells contributes to the development of cardiac fibrosis, ultimately leading to cardiac remodeling. In this study, the effects and molecular mechanisms of celastrol (CEL) on transforming growth factor-β1 (TGF-β1)-induced EndMT in human umbilical vein endothelial (HUVEC-12) cells were investigated. The presented data demonstrated that CEL significantly blocked the morphology change of HUVEC-12 cells induced by TGF-β1 without cell cytotoxicity. In accordance with these findings, CEL blocked TGF-β1-induced EndMT as evidenced by the inhibition of the mesenchymal markers, including collagen I, III, α-SMA, fibronectin mRNA expression, and the increase in the mRNA expression of endothelial cell marker CD31. These changes were also confirmed by double immunofluorescence staining of CD31 and vimentin. The in vitro scratch assay showed that CEL inhibited the migration capacity of the transitioned endothelial cells induced by TGF-β1. Further experiments showed that the beneficial effect of CEL on blocking the EndMT in HUVEC-12 cells was associated with the suppression of the TGF-β1/Smads signalling pathway, which was also confirmed by the inhibition of its downstream transcription factor snail1, twist1, twist2, ZEB1 and ZEB2. These results indicate that CEL blocks TGF-β1-induced EndMT through TGF-β1/Smads signalling pathway and suggest that it may be a feasible therapy for cardiac fibrosis diseases.

Keywords

endothelial-mesenchymal transition / celastrol / endothelium / TGF-β1/Smads signaling pathway / cardiac fibrosis

References

[1] BujakM, RenG, KweonHJ, et al. . Essential role of Smad3 in infarct healing and in the pathogenesis of cardiac remodeling. Circulation, 2007, 116(19): 2127-2138 PMID: 17967775

[2] van AmerongenMJ, Bou-GhariosG, PopaE, et al. . Bone marrow-derived myofibroblasts contribute functionally to scar formation after myocardial infarction. J Pathol, 2008, 214(3): 377-386 PMID: 18095257

[3] KolditzDP, WijffelsMC, BlomNA, et al. . Epicardium-derived cells in development of annulus fibrosis and persistence of accessory pathways. Circulation, 2008, 117(12): 1508-1517 PMID: 18332266

[4] Diaz-FloresL, GutierrezR, MadridJF, et al. . Pericytes. Morphofunction, interactions and pathology in a quiescent and activated mesenchymal cell niche. Histol Histopathol, 2009, 24(7): 909-969

[5] ZeisbergEM, TarnavskiO, ZeisbergM, et al. . Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nat Med, 2007, 13(8): 952-961 PMID: 17660828

[6] ZeisbergM, KalluriR. Fibroblasts emerge via epithelial-mesenchymal transition in chronic kidney fibrosis. Front Biosci, 2008, 13: 6991-6998 PMID: 18508710

[7] LiJ, QuX, YaoJ, et al. . Blockade of endothelial-mesenchymal transition by a Smad3 inhibitor delays the early development of streptozotocin-induced diabetic nephropathy. Diabetes, 2010, 59(10): 2612-2624 PMID: 20682692 PMCID: 3279546

[8] HashimotoN, PhanSH, ImaizumiK, et al. . Endothelial-mesenchymal transition in bleomycin-induced pulmonary fibrosis. Am J Respir Cell Mol Biol, 2010, 43(2): 161-172 PMID: 19767450

[9] WidyantoroB, EmotoN, NakayamaK, et al. . Endothelial cell-derived endothelin-1 promotes cardiac fibrosis in diabetic hearts through stimulation of endothelial-to-mesenchymal transition. Circulation, 2010, 121(22): 2407-2418 PMID: 20497976

[10] MurdochCE, ChaubeyS, ZengL, et al. . Endothelial NADPH oxidase-2 promotes interstitial cardiac fibrosis and diastolic dysfunction through proinflammatory effects and endothelial-mesenchymal transition. J Am Coll Cardiol, 2014, 63(24): 2734-2741 PMID: 24681145

[11] KovacicJC, MercaderN, TorresM, et al. . Epithelial-to-mesenchymal and endothelial-to-mesenchymal transition: from cardiovascular development to disease. Circulation, 2012, 125(14): 1795-1808 PMID: 22492947 PMCID: 3333843

[12] Piera-VelazquezS, MendozaFA, JimenezSA. Endothelial to Mesenchymal transition (EndoMT) in the pathogenesis of human fibrotic diseases. J Clin Med, 2016, 5(4): E45 PMID: 27077889

[13] CalixtoJB, CamposMM, OtukiMF, et al. . Anti-inflammatory compounds of plant origin. Part II. modulation of pro-inflammatory cytokines, chemokines and adhesion molecules. Planta Med, 2004, 70(2): 93-103 PMID: 14994184

[14] LiPP, HeW, YuanPF, et al. . Celastrol induces mitochondria-mediated apoptosis in hepatocellular carcinoma Bel-7402 cells. Am J Chin Med, 2015, 43(1): 137-148 PMID: 25657108

[15] KannaiyanR, ShanmugamMK, SethiG. Molecular targets of celastrol derived from Thunder of God Vine: potential role in the treatment of inflammatory disorders and cancer. Cancer Lett, 2011, 303(1): 9-20 PMID: 21168266

[16] SalminenA, LehtonenM, PaimelaT, et al. . Celastrol: Molecular targets of Thunder God Vine. Biochem Biophys Res Commun, 2010, 394(3): 439-442 PMID: 20226165

[17] YuX, TaoW, JiangF, et al. . Celastrol attenuates hypertension-induced inflammation and oxidative stress in vascular smooth muscle cells via induction of heme oxygenase-1. Am J Hypertens, 2010, 23(8): 895-903 PMID: 20414191

[18] WangYL, LamKK, ChengPY, et al. . Celastrol prevents circulatory failure via induction of heme oxygenase-1 and heat shock protein 70 in endotoxemic rats. J Ethnopharmacol, 2015, 162: 168-175 PMID: 25571843

[19] Porter KeT Na. Cardiac fibroblasts: at the heart of myocardial remodeling. Pharmacol Ther, 2009, 123(2): 255-278 PMID: 19460403

[20] TraversJG, KamalFA, RobbinsJ, et al. . Cardiac fibrosis: the fibroblast awakens. Circ Res, 2016, 118(6): 1021-1040 PMID: 26987915 PMCID: 4800485

[21] HuaJY, ZhangZC, JiangXH, et al. . Relationship between endothelial-to-mesenchymal transition and cardiac fibrosis in acute viral myocarditis. Zhejiang Da Xue Xue Bao Yi Xue Ban (Chinese), 2012, 41(3): 298-304

[22] ZhouH, GuoH, ZongJ, et al. . ATF3 regulates multiple targets and may play a dual role in cardiac hypertrophy and injury. Int J Cardiol, 2014, 174(3): 838-839 PMID: 24794959

[23] GrotendorstGR, RahmanieH, DuncanMR. Combinatorial signaling pathways determine fibroblast proliferation and myofibroblast differentiation. FASEB J, 2004, 18(3): 469-479 PMID: 15003992

[24] ChanEC, PeshavariyaHM, LiuGS, et al. . Nox4 modulates collagen production stimulated by transforming growth factor beta1 in vivo and in vitro. Biochem Biophys Res Commun, 2013, 430(3): 918-925 PMID: 23261430

[25] ChenLX, YangK, SunM, et al. . Fluorofenidone inhibits transforming growth factor-beta1-induced cardiac myofibroblast differentiation. Pharmazie, 2012, 67(5): 452-456 PMID: 22764581

[26] DengYL, XiongXZ, ChengNS. Organ fibrosis inhibited by blocking transforming growth factor-beta signaling via peroxisome proliferator-activated receptor gamma agonists. Hepatobiliary Pancreat Dis Int, 2012, 11(5): 467-478 PMID: 23060391

[27] ZhanCY, TangJH, ZhouDX, et al. . Effects of tanshinone IIA on the transforming growth factor beta1/Smad signaling pathway in rat cardiac fibroblasts. Indian J Pharmacol, 2014, 46(6): 633-638 PMID: 25538336 PMCID: 4264080