Characterization of fibroblasts from hypertrophied right ventricle of pulmonary hypertensive rats
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Pulmonary arterial hypertension (PAH), which is characterized by an elevation of pulmonary arterial resistance, leads to a lethal right heart failure. It is an urgent issue to clarify the pathogenesis of PAH-induced right heart failure. The present study aimed to elucidate the characteristics of cardiac fibroblasts (CFs) isolated from hypertrophied right ventricles of monocrotaline (MCT)-induced PAH model rats. CFs were isolated from the right ventricles of MCT-injected rats (MCT-CFs) and saline-injected control rats (CONT-CFs). Expression of α-smooth muscle actin and collagen type I in MCT-CFs was lower than that in CONT-CFs. On the other hand, proliferation, migration, and matrix metalloproteinase (MMP)-9 production were significantly enhanced in MCT-CFs. In MCT-CFs, phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, c-Jun N-terminal kinase (JNK), and Ca2+/calmodulin-dependent protein kinase (CaMK) II was significantly enhanced. In addition to mRNA expression of Orai1, a Ca2+ release-activated Ca2+ channel, and stromal interaction molecules (STIM) 1, an endoplasmic reticulum Ca2+ sensor, the associated store-operated Ca2+ entry (SOCE) was significantly higher in MCT-CFs than CONT-CFs. Pharmacological inhibition of ERK1/2 pathway prevented the enhanced proliferation of MCT-CFs. The enhanced migration of MCT-CFs was prevented by a pharmacological inhibition of ERK1/2, JNK, CaMKII, or SOCE pathway. The enhanced MMP-9 production in MCT-CFs was prevented by a pharmacological inhibition of ERK1/2, CaMKII, or SOCE pathway but not JNK. The present results suggested that MCT-CFs exhibit proliferative and migratory phenotypes perhaps through multiple signaling pathways. This study for the first time determined the characteristics of CFs isolated from hypertrophied right ventricles of MCT-induced PAH model rats.
KeywordsCardiac fibroblasts Pulmonary arterial hypertension Right heart failure Proliferation Migration
This research was supported by JSPS KAKENHI Grant Number 18J11540 (Grant-in-Aid for JSPS Research Fellow).
Compliance with ethical standards
All animal care and treatments were conducted in accordance with the National Institutes of Health Guide for the Care of Laboratory Animals and the guidelines of the Kitasato University. All animal studies were approved by the President of Kitasato University through the judgment by Institutional Animal Care and Use Committee of Kitasato University (Approval no. 16-043 and 17-085).
- 3.Chen JC, Lin BB, Hu HW, Lin C, Jin WY, Zhang FB, Zhu YA, Lu CJ, Wei XJ, Chen RJ (2014) NGF accelerates cutaneous wound healing by promoting the migration of dermal fibroblasts via the PI3K/Akt-Rac1-JNK and ERK pathways. Biomed Res Int 2014:547187–547113. https://doi.org/10.1155/2014/547187 PubMedPubMedCentralCrossRefGoogle Scholar
- 8.Galie N, Humbert M, Vachiery JL, Gibbs S, Lang I, Torbicki A, Simonneau G, Peacock A, Vonk Noordegraaf A, Beghetti M, Ghofrani A, Gomez Sanchez MA, Hansmann G, Klepetko W, Lancellotti P, Matucci M, McDonagh T, Pierard LA, Trindade PT, Zompatori M, Hoeper M, Aboyans V, Vaz Carneiro A, Achenbach S, Agewall S, Allanore Y, Asteggiano R, Paolo Badano L, Albert Barbera J, Bouvaist H, Bueno H, Byrne RA, Carerj S, Castro G, Erol C, Falk V, Funck-Brentano C, Gorenflo M, Granton J, Iung B, Kiely DG, Kirchhof P, Kjellstrom B, Landmesser U, Lekakis J, Lionis C, Lip GY, Orfanos SE, Park MH, Piepoli MF, Ponikowski P, Revel MP, Rigau D, Rosenkranz S, Voller H, Luis Zamorano J (2016) 2015 ESC/ERS guidelines for the diagnosis and treatment of pulmonary hypertension: the Joint Task Force for the Diagnosis and Treatment of Pulmonary Hypertension of the European Society of Cardiology (ESC) and the European Respiratory Society (ERS): endorsed by: Association for European Paediatric and Congenital Cardiology (AEPC), International Society for Heart and Lung Transplantation (ISHLT). Eur Heart J 37:67–119. https://doi.org/10.1093/eurheartj/ehv317 CrossRefPubMedGoogle Scholar
- 9.Gomez-Arroyo J, Sakagami M, Syed AA, Farkas L, Van Tassell B, Kraskauskas D, Mizuno S, Abbate A, Bogaard HJ, Byron PR, Voelkel NF (2015) Iloprost reverses established fibrosis in experimental right ventricular failure. Eur Respir J 45:449–462. https://doi.org/10.1183/09031936.00188013 CrossRefPubMedGoogle Scholar
- 10.Guo JL, Yu Y, Jia YY, Ma YZ, Zhang BY, Liu PQ, Chen SR, Jiang JM (2014) Transient receptor potential melastatin 7 (TRPM7) contributes to H2O2-induced cardiac fibrosis via mediating Ca(2+) influx and extracellular signal-regulated kinase 1/2 (ERK1/2) activation in cardiac fibroblasts. J Pharmacol Sci 125:184–192CrossRefPubMedGoogle Scholar
- 11.Harada M, Luo X, Qi XY, Tadevosyan A, Maguy A, Ordog B, Ledoux J, Kato T, Naud P, Voigt N, Shi Y, Kamiya K, Murohara T, Kodama I, Tardif JC, Schotten U, Van Wagoner DR, Dobrev D, Nattel S (2012) Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation. Circulation 126:2051–2064. https://doi.org/10.1161/CIRCULATIONAHA.112.121830 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Hoeper MM, Kramer T, Pan Z, Eichstaedt CA, Spiesshoefer J, Benjamin N, Olsson KM, Meyer K, Vizza CD, Vonk-Noordegraaf A, Distler O, Opitz C, Gibbs JSR, Delcroix M, Ghofrani HA, Huscher D, Pittrow D, Rosenkranz S, Grunig E (2017) Mortality in pulmonary arterial hypertension: prediction by the 2015 European pulmonary hypertension guidelines risk stratification model. Eur Respir J 50. doi: https://doi.org/10.1183/13993003.00740-2017
- 14.Hwang YP, Kim HG, Choi JH, Park BH, Jeong MH, Jeong TC, Jeong HG (2011) Acteoside inhibits PMA-induced matrix metalloproteinase-9 expression via CaMK/ERK- and JNK/NF-kappaB-dependent signaling. Mol Nutr Food Res 55(Suppl 1):S103–S116. https://doi.org/10.1002/mnfr.201000336 CrossRefPubMedGoogle Scholar
- 15.Ikeda S, Satoh K, Kikuchi N, Miyata S, Suzuki K, Omura J, Shimizu T, Kobayashi K, Fukumoto Y, Sakata Y, Shimokawa H (2014) Crucial role of rho-kinase in pressure overload-induced right ventricular hypertrophy and dysfunction in mice. Arterioscler Thromb Vasc Biol 34:1260–1271. https://doi.org/10.1161/ATVBAHA.114.303320 CrossRefPubMedGoogle Scholar
- 18.Jessica S, Angele B, Catherine RM, Melanie L, Ana-Maria G, Jean-Pierre B, Frederic P, Marc H, Fabrice A (2018) Ca(2+) handling remodeling and STIM1L/Orai1/TRPC1/TRPC4 upregulation in monocrotaline-induced right ventricular hypertrophy. J Mol Cell Cardiol 118:208–224. https://doi.org/10.1016/j.yjmcc.2018.04.003 CrossRefPubMedGoogle Scholar
- 19.Kimura H, Okubo N, Chosa N, Kyakumoto S, Kamo M, Miura H, Ishisaki A (2013) EGF positively regulates the proliferation and migration, and negatively regulates the myofibroblast differentiation of periodontal ligament-derived endothelial progenitor cells through MEK/ERK- and JNK-dependent signals. Cell Physiol Biochem 32:899–914. https://doi.org/10.1159/000354493 CrossRefPubMedGoogle Scholar
- 21.Lee WT, Lee TH, Cheng CH, Chen KC, Chen YC, Lin CW (2015) Antroquinonol from Antrodia camphorata suppresses breast tumor migration/invasion through inhibiting ERK-AP-1- and AKT-NF-kappaB-dependent MMP-9 and epithelial-mesenchymal transition expressions. Food Chem Toxicol 78:33–41. https://doi.org/10.1016/j.fct.2015.01.012 CrossRefPubMedGoogle Scholar
- 22.Martin TP, Lawan A, Robinson E, Grieve DJ, Plevin R, Paul A, Currie S (2014) Adult cardiac fibroblast proliferation is modulated by calcium/calmodulin-dependent protein kinase II in normal and hypertrophied hearts. Pflugers Arch 466:319–330. https://doi.org/10.1007/s00424-013-1326-9 CrossRefPubMedGoogle Scholar
- 23.Mukhopadhyay S, Munshi HG, Kambhampati S, Sassano A, Platanias LC, Stack MS (2004) Calcium-induced matrix metalloproteinase 9 gene expression is differentially regulated by ERK1/2 and p38 MAPK in oral keratinocytes and oral squamous cell carcinoma. J Biol Chem 279:33139–33146. https://doi.org/10.1074/jbc.M405194200 CrossRefPubMedGoogle Scholar
- 29.Ross GR, Bajwa T Jr, Edwards S, Emelyanova L, Rizvi F, Holmuhamedov EL, Werner P, Downey FX, Tajik AJ, Jahangir A (2017) Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart. Biol Open 6:326–332. https://doi.org/10.1242/bio.022632 CrossRefPubMedPubMedCentralGoogle Scholar
- 31.Siddesha JM, Valente AJ, Sakamuri SS, Gardner JD, Delafontaine P, Noda M, Chandrasekar B (2014) Acetylsalicylic acid inhibits IL-18-induced cardiac fibroblast migration through the induction of RECK. J Cell Physiol 229:845–855. https://doi.org/10.1002/jcp.24511 CrossRefPubMedPubMedCentralGoogle Scholar
- 37.Zhang X, Zhang T, Wu J, Yu X, Zheng D, Yang F, Li T, Wang L, Zhao Y, Dong S, Zhong X, Fu S, Xu CQ, Lu F, Zhang WH (2014) Calcium sensing receptor promotes cardiac fibroblast proliferation and extracellular matrix secretion. Cell Physiol Biochem 33:557–568. https://doi.org/10.1159/000358634 CrossRefPubMedGoogle Scholar