Skip to main content

Advertisement

SpringerLink
Log in

Characterization of fibroblasts from hypertrophied right ventricle of pulmonary hypertensive rats

  • Signaling and cell physiology
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Benza RL, Miller DP, Barst RJ, Badesch DB, Frost AE, McGoon MD (2012) An evaluation of long-term survival from time of diagnosis in pulmonary arterial hypertension from the REVEAL Registry. Chest 142:448–456. https://doi.org/10.1378/chest.11-1460

    Article  PubMed  Google Scholar 

  2. Cai J, Du S, Wang H, Xin B, Wang J, Shen W, Wei W, Guo Z, Shen X (2017) Tenascin-C induces migration and invasion through JNK/c-Jun signalling in pancreatic cancer. Oncotarget 8:74406–74422. https://doi.org/10.18632/oncotarget.20160

    Article  PubMed  PubMed Central  Google Scholar 

  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

    Article  PubMed  PubMed Central  Google Scholar 

  4. Dobaczewski M, Chen W, Frangogiannis NG (2011) Transforming growth factor (TGF)-beta signaling in cardiac remodeling. J Mol Cell Cardiol 51:600–606. https://doi.org/10.1016/j.yjmcc.2010.10.033

    Article  PubMed  CAS  Google Scholar 

  5. Du J, Xie J, Zhang Z, Tsujikawa H, Fusco D, Silverman D, Liang B, Yue L (2010) TRPM7-mediated Ca2+ signals confer fibrogenesis in human atrial fibrillation. Circ Res 106:992–1003. https://doi.org/10.1161/CIRCRESAHA.109.206771

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Dzeshka MS, Lip GY, Snezhitskiy V, Shantsila E (2015) Cardiac fibrosis in patients with atrial fibrillation: mechanisms and clinical implications. J Am Coll Cardiol 66:943–959. https://doi.org/10.1016/j.jacc.2015.06.1313

    Article  PubMed  Google Scholar 

  7. Frangogiannis NG (2015) Interleukin-1 in cardiac injury, repair, and remodeling: pathophysiologic and translational concepts. Discoveries (Craiova) 3:e41. https://doi.org/10.15190/d.2015.33

    Article  Google 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

    Article  PubMed  Google 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

    Article  PubMed  CAS  Google 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 125:184–192

    Article  PubMed  CAS  Google 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

    Article  PubMed  PubMed Central  CAS  Google 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

  13. Hu J, Wang X, Wei SM, Tang YH, Zhou Q, Huang CX (2016) Activin A stimulates the proliferation and differentiation of cardiac fibroblasts via the ERK1/2 and p38-MAPK pathways. Eur J Pharmacol 789:319–327. https://doi.org/10.1016/j.ejphar.2016.07.053

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google Scholar 

  16. Imoto K, Kumatani S, Okada M, Yamawaki H (2016) Endostatin is protective against monocrotaline-induced right heart disease through the inhibition of T-type Ca(2+) channel. Pflugers Arch 468:1259–1270. https://doi.org/10.1007/s00424-016-1810-0

    Article  PubMed  CAS  Google Scholar 

  17. Imoto K, Okada M, Yamawaki H (2017) Expression profile of matricellular proteins in hypertrophied right ventricle of monocrotaline-induced pulmonary hypertensive rats. J Vet Med Sci 79:1096–1102. https://doi.org/10.1292/jvms.17-0053

    Article  PubMed  PubMed Central  Google 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

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google Scholar 

  20. Lee YC, Chang AY, Lin-Feng MH, Tsou WI, Chiang IH, Lai MZ (2012) Paxillin phosphorylation by JNK and p38 is required for NFAT activation. Eur J Immunol 42:2165–2175. https://doi.org/10.1002/eji.201142192

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google Scholar 

  24. Ogawa A, Satoh T, Tamura Y, Fukuda K, Matsubara H (2017) Survival of Japanese patients with idiopathic/heritable pulmonary arterial hypertension. Am J Cardiol 119:1479–1484. https://doi.org/10.1016/j.amjcard.2017.01.015

    Article  PubMed  Google Scholar 

  25. Okada M, Kikuzuki R, Harada T, Hori Y, Yamawaki H, Hara Y (2008) Captopril attenuates matrix metalloproteinase-2 and -9 in monocrotaline-induced right ventricular hypertrophy in rats. J Pharmacol Sci 108:487–494

    Article  PubMed  CAS  Google Scholar 

  26. Okada M, Suzuki A, Yamawaki H, Hara Y (2013) Levosimendan inhibits interleukin-1beta-induced cell migration and MMP-9 secretion in rat cardiac fibroblasts. Eur J Pharmacol 718:332–339. https://doi.org/10.1016/j.ejphar.2013.08.013

    Article  PubMed  CAS  Google Scholar 

  27. Okada M, Oba Y, Yamawaki H (2015) Endostatin stimulates proliferation and migration of adult rat cardiac fibroblasts through PI3K/Akt pathway. Eur J Pharmacol 750:20–26. https://doi.org/10.1016/j.ejphar.2015.01.019

    Article  PubMed  CAS  Google Scholar 

  28. Okada M, Murata N, Yamawaki H (2017) Canstatin stimulates migration of rat cardiac fibroblasts via secretion of matrix metalloproteinase-2. Am J Physiol Cell Physiol 312:C199–C208. https://doi.org/10.1152/ajpcell.00329.2015

    Article  PubMed  Google 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

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. Saxena A, Chen W, Su Y, Rai V, Uche OU, Li N, Frangogiannis NG (2013) IL-1 induces proinflammatory leukocyte infiltration and regulates fibroblast phenotype in the infarcted myocardium. J Immunol 191:4838–4848. https://doi.org/10.4049/jimmunol.1300725

    Article  PubMed  CAS  Google 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

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Squires CE, Escobar GP, Payne JF, Leonardi RA, Goshorn DK, Sheats NJ, Mains IM, Mingoia JT, Flack EC, Lindsey ML (2005) Altered fibroblast function following myocardial infarction. J Mol Cell Cardiol 39:699–707. https://doi.org/10.1016/j.yjmcc.2005.07.008

    Article  PubMed  CAS  Google Scholar 

  33. Steiropoulos P, Trakada G, Bouros D (2008) Current pharmacological treatment of pulmonary arterial hypertension. Curr Clin Pharmacol 3:11–19

    Article  PubMed  CAS  Google Scholar 

  34. Usui T, Morita T, Okada M, Yamawaki H (2014) Histone deacetylase 4 controls neointimal hyperplasia via stimulating proliferation and migration of vascular smooth muscle cells. Hypertension 63:397–403. https://doi.org/10.1161/HYPERTENSIONAHA.113.01843

    Article  PubMed  CAS  Google Scholar 

  35. Wu CY, Hsieh HL, Sun CC, Yang CM (2009) IL-1beta induces MMP-9 expression via a Ca2+-dependent CaMKII/JNK/c-JUN cascade in rat brain astrocytes. Glia 57:1775–1789. https://doi.org/10.1002/glia.20890

    Article  PubMed  Google Scholar 

  36. Xie Z, Singh M, Singh K (2004) Differential regulation of matrix metalloproteinase-2 and -9 expression and activity in adult rat cardiac fibroblasts in response to interleukin-1beta. J Biol Chem 279:39513–39519. https://doi.org/10.1074/jbc.M405844200

    Article  PubMed  CAS  Google 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

    Article  PubMed  CAS  Google Scholar 

Download references

Funding

This research was supported by JSPS KAKENHI Grant Number 18J11540 (Grant-in-Aid for JSPS Research Fellow).

Author information

Authors and Affiliations

  1. Laboratory of Veterinary Pharmacology, School of Veterinary Medicine, Kitasato University, Higashi 23 bancho 35-1, Towada City, Aomori, 034-8628, Japan

    Keisuke Imoto, Muneyoshi Okada & Hideyuki Yamawaki

Authors
  1. Keisuke Imoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Muneyoshi Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hideyuki Yamawaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Muneyoshi Okada.

Ethics declarations

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).

Electronic supplementary material

ESM 1

(PDF 163 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Imoto, K., Okada, M. & Yamawaki, H. Characterization of fibroblasts from hypertrophied right ventricle of pulmonary hypertensive rats. Pflugers Arch - Eur J Physiol 470, 1405–1417 (2018). https://doi.org/10.1007/s00424-018-2158-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-018-2158-4

Keywords

Search

Navigation