Skip to main content

Advertisement

SpringerLink
Log in

CTRP15 derived from cardiac myocytes attenuates TGFβ1-induced fibrotic response in cardiac fibroblasts

  • Original Article
  • Published:
Cardiovascular Drugs and Therapy Aims and scope Submit manuscript

Abstract

Purpose

Cardiac fibrosis is characterized by net accumulation of extracellular matrix (ECM) components in the  myocardium and facilitates the development of heart failure. C1q/tumor necrosis factor-related protein 15 (CTRP15) is a novel member of the CTRP family, and its gene expression is detected in adult mouse hearts. The present study was performed to determine the effect of CTRP15 on pressure overload-induced fibrotic remodeling.

Methods

Mice were subjected to transverse aortic constriction (TAC) surgery, and adeno-associated virus serotype 9 (AAV9)-carrying mouse CTRP15 gene was injected into mice to achieve CTRP15 overexpression in the myocardium. Adenovirus carrying the gene encoding CTRP15 or small interfering RNA (siRNA) of interest was infected into cultured neonatal mouse ventricular cardiomyocytes (NMVCs) or cardiac fibroblasts (CFs). Gene expression was measured by quantitative real-time PCR, and protein expression and distribution were determined by Western blotting, immunocytochemistry, and immunofluorescence staining.

Results

CTRP15 was predominantly produced by cardiac myocytes. CTRP15 expression in the left ventricles was downregulated in mice that underwent TAC. AAV9-mediated CTRP15 overexpression alleviated ventricular remodeling and dysfunction in the pressure-overloaded mice. Treatment of CFs with recombinant CTRP15 or the conditioned medium containing CTRP15 inhibited transforming growth factor (TGF)-β1-induced Smad3 activation and myofibroblast differentiation. CTRP15 increased phosphorylation of insulin receptor (IR), insulin receptor substrate-1 (IRS-1), and Akt. Blockade of IR/IRS-1/Akt pathway reversed the inhibitory effect of CTRP15 on TGF-β1-induced Smad3 activation.

Conclusion

CTRP15 exerts an anti-fibrotic effect on pressure overload-induced cardiac remodeling. The activation of IR/IRS-1/Akt pathway contributes to the anti-fibrotic effect of CTRP15 through targeting Smad3.

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. Swynghedauw B. Molecular mechanisms of myocardial remodeling. Physiol Rev. 1999;79(1):215–62.

    CAS  PubMed  Google Scholar 

  2. Creemers EE, Pinto YM. Molecular mechanisms that control interstitial fibrosis in the pressure-overloaded heart. Cardiovasc Res. 2011;89(2):265–72.

    CAS  PubMed  Google Scholar 

  3. Segura AM, Frazier OH, Buja LM. Fibrosis and heart failure. Heart Fail Rev. 2014;19(2):173–85.

    CAS  PubMed  Google Scholar 

  4. Sivakumar P, Gupta S, Sarkar S, Sen S. Upregulation of lysyl oxidase and MMPs during cardiac remodeling in human dilated cardiomyopathy. Mol Cell Biochem. 2008;307(1–2):159–67.

    CAS  PubMed  Google Scholar 

  5. Espira L, Czubryt MP. Emerging concepts in cardiac matrix biology. Can J Physiol Pharmacol. 2009;87(12):996–1008.

    CAS  PubMed  Google Scholar 

  6. Edgley AJ, Krum H, Kelly DJ. Targeting fibrosis for the treatment of heart failure: a role for transforming growth factor-beta. Cardiovasc Ther. 2012;30(1):e30–40.

    CAS  PubMed  Google Scholar 

  7. Fan D, Takawale A, Lee J, Kassiri Z. Cardiac fibroblasts, fibrosis and extracellular matrix remodeling in heart disease. Fibrogenesis Tissue Repair. 2012;5(1):15.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Leask A. TGFbeta, cardiac fibroblasts, and the fibrotic response. Cardiovasc Res. 2007;74(2):207–12.

    CAS  PubMed  Google Scholar 

  9. Krenning G, Zeisberg EM, Kalluri R. The origin of fibroblasts and mechanism of cardiac fibrosis. J Cell Physiol. 2010;225(3):631–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Takeda N, Manabe I, Uchino Y, Eguchi K, Matsumoto S, Nishimura S, et al. Cardiac fibroblasts are essential for the adaptive response of the murine heart to pressure overload. J Clin Invest. 2010;120(1):254–65.

    CAS  PubMed  Google Scholar 

  11. Weber KT, Sun Y, Bhattacharya SK, Ahokas RA, Gerling IC. Myofibroblast-mediated mechanisms of pathological remodelling of the heart. Nat Rev Cardiol. 2013;10(1):15–26.

    CAS  PubMed  Google Scholar 

  12. Swaney JS, Roth DM, Olson ER, Naugle JE, Meszaros JG, Insel PA. Inhibition of cardiac myofibroblast formation and collagen synthesis by activation and overexpression of adenylyl cyclase. Proc Natl Acad Sci U S A. 2005;102(2):437–42.

    CAS  PubMed  Google Scholar 

  13. Shi Q, Liu X, Bai Y, Cui C, Li J, Li Y, et al. In vitro effects of pirfenidone on cardiac fibroblasts: proliferation, myofibroblast differentiation, migration and cytokine secretion. PLoS One. 2011;6(11):e28134.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Stempien-Otero A, Kim DH, Davis J. Molecular networks underlying myofibroblast fate and fibrosis. J Mol Cell Cardiol. 2016;97:153–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Schaffler A, Buechler C. CTRP family: linking immunity to metabolism. Trends Endocrinol Metab. 2012;23(4):194–204.

    PubMed  Google Scholar 

  16. Seldin MM, Peterson JM, Byerly MS, Wei Z, Wong GW. Myonectin (CTRP15), a novel myokine that links skeletal muscle to systemic lipid homeostasis. J Biol Chem. 2012;287(15):11968–80.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Seldin MM, Lei X, Tan SY, Stanson KP, Wei Z, Wong GW. Skeletal muscle-derived myonectin activates the mammalian target of rapamycin (mTOR) pathway to suppress autophagy in liver. J Biol Chem. 2013;288(50):36073–82.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Kautz L, Jung G, Valore EV, Rivella S, Nemeth E, Ganz T. Identification of erythroferrone as an erythroid regulator of iron metabolism. Nat Genet. 2014;46(7):678–84.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Jiang X, Gao M, Chen Y, Liu J, Qi S, Ma J, et al. EPO-dependent induction of erythroferrone drives hepcidin suppression and systematic iron absorption under phenylhydrazine-induced hemolytic anemia. Blood Cells Mol Dis. 2016;58:45–51.

    CAS  PubMed  Google Scholar 

  20. Latour C, Wlodarczyk MF, Jung G, Gineste A, Blanchard N, Ganz T, et al. Erythroferrone contributes to hepcidin repression in a mouse model of malarial anemia. Haematologica. 2017;102(1):60–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Otaka N, Shibata R, Ohashi K, Uemura Y, Kambara T, Enomoto T, et al. Myonectin is an exercise-induced Myokine that protects the heart from ischemia-reperfusion injury. Circ Res. 2018;123(12):1326–38.

    CAS  PubMed  Google Scholar 

  22. Yuan Y, Lau WB, Su H, Sun Y, Yi W, Du Y, et al. C1q-TNF-related protein-9, a novel cardioprotective cardiokine, requires proteolytic cleavage to generate a biologically active globular domain isoform. Am J Physiol Endocrinol Metab. 2015;308(10):E891–8.

    PubMed  PubMed Central  Google Scholar 

  23. Zhang CL, Zhao Q, Liang H, Qiao X, Wang JY, Wu D, et al. Cartilage intermediate layer protein-1 alleviates pressure overload-induced cardiac fibrosis via interfering TGF-beta1 signaling. J Mol Cell Cardiol. 2018;116:135–44.

    CAS  PubMed  Google Scholar 

  24. Sahn DJ, DeMaria A, Kisslo J, Weyman A. Recommendations regarding quantitation in M-mode echocardiography: results of a survey of echocardiographic measurements. Circulation. 1978;58(6):1072–83.

    CAS  PubMed  Google Scholar 

  25. Zhou YY, Wang SQ, Zhu WZ, Chruscinski A, Kobilka BK, Ziman B, et al. Culture and adenoviral infection of adult mouse cardiac myocytes: methods for cellular genetic physiology. Am J Physiol Heart Circ Physiol. 2000;279(1):H429–36.

    CAS  PubMed  Google Scholar 

  26. Chen R, Feng Y, Wu J, Song Y, Li H, Shen Q, et al. Metformin attenuates angiotensin II-induced TGFbeta1 expression by targeting hepatocyte nuclear factor-4-alpha. Br J Pharmacol. 2018;175(8):1217–29.

    CAS  PubMed  Google Scholar 

  27. Ehler E, Moore-Morris T, Lange S. Isolation and culture of neonatal mouse cardiomyocytes. J Vis Exp. 2013;79:e50154.

    Google Scholar 

  28. Pare B, Deschenes LT, Pouliot R, Dupre N, Gros-Louis F. An optimized approach to recover secreted proteins from fibroblast conditioned-media for sSecretomic analysis. Front Cell Neurosci. 2016;10:70.

    PubMed  PubMed Central  Google Scholar 

  29. Kumar S, Seqqat R, Chigurupati S, Kumar R, Baker KM, Young D, et al. Inhibition of nuclear factor kappaB regresses cardiac hypertrophy by modulating the expression of extracellular matrix and adhesion molecules. Free Radic Biol Med. 2011;50(1):206–15.

    CAS  PubMed  Google Scholar 

  30. Kong P, Christia P, Frangogiannis NG. The pathogenesis of cardiac fibrosis. Cell Mol Life Sci. 2014;71(4):549–74.

    CAS  PubMed  Google Scholar 

  31. Remy I, Montmarquette A, Michnick SW. PKB/Akt modulates TGF-beta signalling through a direct interaction with Smad3. Nat Cell Biol. 2004;6(4):358–65.

    CAS  PubMed  Google Scholar 

  32. Yuasa D, Ohashi K, Shibata R, Mizutani N, Kataoka Y, Kambara T, et al. C1q/TNF-related protein-1 functions to protect against acute ischemic injury in the heart. FASEB J. 2016;30(3):1065–75.

    CAS  PubMed  Google Scholar 

  33. Wu D, Lei H, Wang JY, Zhang CL, Feng H, Fu FY, et al. CTRP3 attenuates post-infarct cardiac fibrosis by targeting Smad3 activation and inhibiting myofibroblast differentiation. J Mol Med (Berl). 2015;93(12):1311–25.

    CAS  Google Scholar 

  34. Lei H, Wu D, Wang JY, Li L, Zhang CL, Feng H, et al. C1q/tumor necrosis factor-related protein-6 attenuates post-infarct cardiac fibrosis by targeting RhoA/MRTF-A pathway and inhibiting myofibroblast differentiation. Basic Res Cardiol. 2015;110(4):35.

    PubMed  Google Scholar 

  35. Kambara T, Shibata R, Ohashi K, Matsuo K, Hiramatsu-Ito M, Enomoto T, et al. C1q/tumor necrosis factor-related protein 9 protects against acute myocardial injury through an adiponectin receptor I-AMPK-dependent mechanism. Mol Cell Biol. 2015;35(12):2173–85.

    PubMed  PubMed Central  Google Scholar 

  36. Sun Y, Yi W, Yuan Y, Lau WB, Yi D, Wang X, et al. C1q/tumor necrosis factor-related protein-9, a novel adipocyte-derived cytokine, attenuates adverse remodeling in the ischemic mouse heart via protein kinase a activation. Circulation. 2013;128(11 Suppl 1):S113–20.

    CAS  PubMed  Google Scholar 

  37. Yuan YP, Ma ZG, Zhang X, Xu SC, Zeng XF, Yang Z, et al. CTRP3 protected against doxorubicin-induced cardiac dysfunction, inflammation and cell death via activation of Sirt1. J Mol Cell Cardiol. 2018;114:38–47.

    CAS  PubMed  Google Scholar 

  38. Wei WY, Ma ZG, Zhang N, Xu SC, Yuan YP, Zeng XF, et al. Overexpression of CTRP3 protects against sepsis-induced myocardial dysfunction in mice. Mol Cell Endocrinol. 2018;476:27–36.

    CAS  PubMed  Google Scholar 

  39. Ma ZG, Yuan YP, Xu SC, Wei WY, Xu CR, Zhang X, et al. CTRP3 attenuates cardiac dysfunction, inflammation, oxidative stress and cell death in diabetic cardiomyopathy in rats. Diabetologia. 2017;60(6):1126–37.

    CAS  PubMed  Google Scholar 

  40. Bai S, Cheng L, Yang Y, Fan C, Zhao D, Qin Z, et al. C1q/TNF-related protein 9 protects diabetic rat heart against ischemia reperfusion injury: role of endoplasmic reticulum stress. Oxidative Med Cell Longev. 2016;2016:1902025.

    Google Scholar 

  41. Wang J, Chen H, Seth A, McCulloch CA. Mechanical force regulation of myofibroblast differentiation in cardiac fibroblasts. Am J Physiol Heart Circ Physiol. 2003;285(5):H1871–81.

    CAS  PubMed  Google Scholar 

  42. Hinz B. The myofibroblast: paradigm for a mechanically active cell. J Biomech. 2010;43(1):146–55.

    PubMed  Google Scholar 

  43. Pichler M, Rainer PP, Schauer S, Hoefler G. Cardiac fibrosis in human transplanted hearts is mainly driven by cells of intracardiac origin. J Am Coll Cardiol. 2012;59(11):1008–16.

    PubMed  Google Scholar 

  44. Kambara T, Ohashi K, Shibata R, Ogura Y, Maruyama S, Enomoto T, et al. CTRP9 protein protects against myocardial injury following ischemia-reperfusion through AMP-activated protein kinase (AMPK)-dependent mechanism. J Biol Chem. 2012;287(23):18965–73.

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Han CY, Koo JH, Kim SH, Gardenghi S, Rivella S, Strnad P, et al. Hepcidin inhibits Smad3 phosphorylation in hepatic stellate cells by impeding ferroportin-mediated regulation of Akt. Nat Commun. 2016;7:13817.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

Authors acknowledge Dr. Yao Song (Peking University Third Hospital, Beijing, China) for technical support in echocardiographic examination. We also acknowledge Dr. Zhen-Zhen Chen (Peking University School of Basic Medical Sciences, Beijing, China) for helping to inject AAV9 in animal experiments.

Funding

This work is supported by grants from the National Natural Science Foundation of China (81770225 and 81370192).

Author information

Authors and Affiliations

  1. Department of Physiology and Pathophysiology, Peking University School of Basic Medical Sciences; Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, 100191, China

    Qian Zhao, Cheng-Lin Zhang, Ruo-Lan Xiang, Li-Ling Wu & Li Li

Authors
  1. Qian Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cheng-Lin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ruo-Lan Xiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li-Ling Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Li Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Li Li.

Ethics declarations

All experimental procedures were conducted with the approval of the Ethics Committee of Animal Research, Peking University Health Science Center, and the investigation conformed to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85–23, revised 1996).

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Qian Zhao and Cheng-Lin Zhang contribute equally to this work.

Electronic supplementary material

ESM 1

(DOCX 191 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhao, Q., Zhang, CL., Xiang, RL. et al. CTRP15 derived from cardiac myocytes attenuates TGFβ1-induced fibrotic response in cardiac fibroblasts. Cardiovasc Drugs Ther 34, 591–604 (2020). https://doi.org/10.1007/s10557-020-06970-6

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10557-020-06970-6

Keywords

Search

Navigation