Therapeutic effects of the selective farnesoid X receptor agonist obeticholic acid in a monocrotaline-induced pulmonary hypertension rat model

  • P. Comeglio
  • S. Filippi
  • E. Sarchielli
  • A. Morelli
  • I. Cellai
  • C. Corno
  • L. Adorini
  • G. B. Vannelli
  • M. Maggi
  • L. VignozziEmail author
Original Article



Activation of the farnesoid X receptor (FXR), a member of the nuclear receptor steroid superfamily, leads to anti-inflammatory and anti-fibrotic effects in several tissues, including the lung. We have recently demonstrated a protective effect of the farnesoid X receptor (FXR) agonist obeticholic acid (OCA) in rat models of monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) and bleomycin-induced pulmonary fibrosis. The aim of the present study was to investigate whether the positive effects of OCA treatment could be exerted also in established MCT-induced PAH, i.e., starting treatment 2 weeks after MCT administration.


Rats with MCT-induced PAH were treated, 2 weeks after MCT administration, with OCA or tadalafil for two additional weeks. Pulmonary functional tests were performed at week 2 (before treatment) and four (end of treatment). At the same time points, lung morphological features and expression profile of genes related to smooth muscle relaxation/contraction and tissue remodeling were also assessed.


2 weeks after MCT-induced injury, the treadmill resistance (a functional parameter related to pulmonary hypertension) was significantly decreased. At the same time point, we observed right ventricular hypertrophy and vascular remodeling, with upregulation of genes related to inflammation. At week 4, we observed a further worsening of the functional and morphological parameters, accompanied by dysregulation of inflammatory and extracellular matrix markers mRNA expression. Administration of OCA (3 or 10 mg/kg/day), starting 2 weeks after MCT-induced injury, significantly improved pulmonary function, effectively normalizing the exercise capacity. OCA also reverted most of the lung alterations, with a significant reduction of lung vascular wall thickness, right ventricular hypertrophy, and restoration of the local balance between relaxant and contractile pathways. Markers of remodeling pathways were also normalized by OCA treatment. Notably, results with OCA treatment were similar, or even superior, to those obtained with tadalafil, a recently approved treatment for pulmonary hypertension.


The results of this study demonstrate a significant therapeutic effect of OCA in established MCT-induced PAH, improving exercise capacity associated with reduction of right ventricular hypertrophy and lung vascular remodeling. Thus, OCA dosing in a therapeutic protocol restores the balance between relaxant and contractile pathways in the lung, promoting cardiopulmonary protective actions in MCT-induced PAH.


Monocrotaline Pulmonary hypertension Inflammation Farnesoid X receptor Obeticholic acid 



This study has been supported by a scientific Grant from Intercept Pharmaceuticals (New York, NY).

Compliance with ethical standards

Conflict of interest

PC, SF, ES, AM, IC, CC, GBV, MM and LV have no conflicts of interest. LA is a scientific consultant for Intercept Pharmaceuticals.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Informed consent

This article does not contain any studies with human participants performed by any of the authors.


  1. 1.
    Li T, Chiang JY (2014) Bile acid signaling in metabolic disease and drug therapy. Pharmacol Rev 66:948–983CrossRefGoogle Scholar
  2. 2.
    Copple BL, Li T (2016) Pharmacology of bile acid receptors: evolution of bile acids from simple detergents to complex signaling molecules. Pharmacol Res 104:9–21CrossRefGoogle Scholar
  3. 3.
    Cariou B, van Harmelen K, Duran-Sandoval D et al (2006) The farnesoid X receptor modulates adiposity and peripheral insulin sensitivity in mice. J Biol Chem 281:11039–11049CrossRefGoogle Scholar
  4. 4.
    Houten SM, Volle DH, Cummins CL et al (2007) In vivo imaging of farnesoid X receptor activity reveals the ileum as the primary bile acid signaling tissue. Mol Endocrinol 21:1312–1323CrossRefGoogle Scholar
  5. 5.
    Schote AB, Turner JD, Schiltz J, Muller CP (2007) Nuclear receptors in human immune cells: expression and correlations. Mol Immunol 44:1436–1445CrossRefGoogle Scholar
  6. 6.
    Higashiyama H, Kinoshita M, Asano S (2008) Immunolocalization of farnesoid X receptor (FXR) in mouse tissues using tissue microarray. Acta Histochem 110:86–93CrossRefGoogle Scholar
  7. 7.
    Lefebvre P, Cariou B, Lien F et al (2009) Role of bile acids and bile acid receptors in metabolic regulation. Physiol Rev 89:147–191CrossRefGoogle Scholar
  8. 8.
    Popescu IR, Helleboid-Chapman A, Lucas A et al (2010) The nuclear receptor FXR is expressed in pancreatic beta-cells and protects human islets from lipotoxicity. FEBS Lett 584:2845–2851CrossRefGoogle Scholar
  9. 9.
    Ali AH, Carey EJ, Lindor KD (2015) Recent advances in the development of farnesoid X receptor agonists. Ann Transl Med 3:5PubMedPubMedCentralGoogle Scholar
  10. 10.
    Ye L, Jiang Y, Zuo X (2015) Farnesoid-X-receptor expression in monocrotaline-induced pulmonary arterial hypertension and right heart failure. Biochem Biophys Res Commun 467:164–170CrossRefGoogle Scholar
  11. 11.
    Comeglio P, Filippi S, Sarchielli E et al (2017) Anti-fibrotic effects of chronic treatment with the selective FXR agonist obeticholic acid in the bleomycin-induced rat model of pulmonary fibrosis. J Steroid Biochem Mol Biol 168:26–37CrossRefGoogle Scholar
  12. 12.
    He F, Li J, Mu Y et al (2006) Downregulation of endothelin-1 by farnesoid X receptor in vascular endothelial cells. Circ Res 98:192–199CrossRefGoogle Scholar
  13. 13.
    Hendrick SM, Mroz MS, Greene CM et al (2014) Bile acids stimulate chloride secretion through CFTR and calcium-activated Cl- channels in Calu-3 airway epithelial cells. Am J Physiol Lung Cell Mol Physiol 307:407–418CrossRefGoogle Scholar
  14. 14.
    Vignozzi L, Morelli A, Cellai I et al (2017) Cardiopulmonary protective effects of the selective FXR agonist obeticholic acid in the rat model of monocrotaline-induced pulmonary hypertension. J Steroid Biochem Mol Biol 165:277–292CrossRefGoogle Scholar
  15. 15.
    Shaik FB, Panati K, Narasimha VR, Narala VR (2015) Chenodeoxycholic acid attenuates ovalbumin-induced airway inflammation in murine model of asthma by inhibiting the T(H)2 cytokines. Biochem Biophys Res Commun 463(4):600–605CrossRefGoogle Scholar
  16. 16.
    Zhang L, Li T, Yu D et al (2012) FXR protects lung from lipopolysaccharide-induced acute injury. Mol Endocrinol 26:27–36CrossRefGoogle Scholar
  17. 17.
    Pellicciari R, Fiorucci S, Camaioni E et al (2002) 6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), a potent and selective FXR agonist endowed with anticholestatic activity. J Med Chem 45:3569–3572CrossRefGoogle Scholar
  18. 18.
    Markham A, Keam SJ (2016) Obeticholic acid: First global approval. Drugs 76(12):1221–1226CrossRefGoogle Scholar
  19. 19.
    Hirschfield GM, Mason A, Luketic V et al (2015) Efficacy of obeticholic acid in patients with primary biliary cirrhosis and inadequate response to ursodeoxycholic acid. Gastroenterology 148:751–761CrossRefGoogle Scholar
  20. 20.
    Nevens F, Andreone P, Mazzella G et al (2016) A placebo-controlled trial of obeticholic acid in primary biliary cholangitis. N Engl J Med 375(7):631–643CrossRefGoogle Scholar
  21. 21.
    Neuschwander-Tetri BA, Loomba R, Sanyal AJ et al (2015) Farnesoid X nuclear receptor ligand obeticholic acid for non-cirrhotic, non-alcoholic steatohepatitis (FLINT): a multicentre, randomised, placebo-controlled trial. Lancet 385:956–965CrossRefGoogle Scholar
  22. 22.
    Mudaliar S, Henry RR, Sanyal AJ et al (2013) Efficacy and safety of the farnesoid X receptor agonist obeticholic acid in patients with type 2 diabetes and nonalcoholic fatty liver disease. Gastroenterology 145:574–582CrossRefGoogle Scholar
  23. 23.
    Wang XX, Jiang T, Shen Y et al (2010) Diabetic nephropathy is accelerated by farnesoid X receptor deficiency and inhibited by farnesoid X receptor activation in a type 1 diabetes model. Diabetes 59:2916–2927CrossRefGoogle Scholar
  24. 24.
    Vignozzi L, Morelli A, Filippi S et al (2011) Farnesoid X receptor activation improves erectile function in animal models of metabolic syndrome and diabetes. J Sex Med 8(1):57–77CrossRefGoogle Scholar
  25. 25.
    Adorini L, Pruzanski M, Shapiro D (2012) Farnesoid X receptor targeting to treat nonalcoholic steatohepatitis. Drug Discov Today 17:988–997CrossRefGoogle Scholar
  26. 26.
    Vignozzi L, Filippi S, Comeglio P et al (2014) Nonalcoholic steatohepatitis as a novel player in metabolic syndrome-induced erectile dysfunction: an experimental study in the rabbit. Mol Cell Endocrinol 384:143–154CrossRefGoogle Scholar
  27. 27.
    Zhou B, Feng B, Qin Z et al (2016) Activation of farnesoid X receptor downregulates visfatin and attenuates diabetic nephropathy. Mol Cell Endocrinol 419:72–82CrossRefGoogle Scholar
  28. 28.
    Rabinovitch M, Guignabert C, Humbert M, Nicolls MR (2014) Inflammation and immunity in the pathogenesis of pulmonary arterial hypertension. Circ Res 115:165–175CrossRefGoogle Scholar
  29. 29.
    Guazzi M, Phillips SA, Arena R, Lavie CJ (2015) Endothelial dysfunction and lung capillary injury in cardiovascular diseases. Prog Cardiovasc Dis 57:454–462CrossRefGoogle Scholar
  30. 30.
    Latus H, Delhaas T, Schranz D, Apitz C (2015) Treatment of pulmonary arterial hypertension in children. Nat Rev Cardiol 12:244–254CrossRefGoogle Scholar
  31. 31.
    Lang M, Kojonazarov B, Tian X et al (2012) The soluble guanylate cyclase stimulator riociguat ameliorates pulmonary hypertension induced by hypoxia and SU5416 in rats. PLoS ONE 7:e43433CrossRefGoogle Scholar
  32. 32.
    Malenfant S, Neyron AS, Paulin R et al (2013) Signal transduction in the development of pulmonary arterial hypertension. Pulm Circ 3:278–293CrossRefGoogle Scholar
  33. 33.
    Jing ZC, Parikh K, Pulido T et al (2013) Efficacy and safety of oral treprostinil monotherapy for the treatment of pulmonary arterial hypertension: a randomized, controlled trial. Circulation 127:624–633CrossRefGoogle Scholar
  34. 34.
    Sitbon O, Channick R, Chin KM et al (2015) Selexipag for the treatment of pulmonary arterial hypertension. N Engl J Med 373:2522–2533CrossRefGoogle Scholar
  35. 35.
    McLaughlin VV, Benza RL, Rubin LJ et al (2010) Addition of inhaled treprostinil to oral therapy for pulmonary arterial hypertension: a randomized controlled clinical trial. J Am Coll Cardiol 55:1915–1922CrossRefGoogle Scholar
  36. 36.
    Ataya A, Cope J, Alnuaimat H (2016) A review of targeted pulmonary arterial hypertension-specific pharmacotherapy. J Clin Med 5(12):E114CrossRefGoogle Scholar
  37. 37.
    Stenmark KR, Meyrick B, Galie N et al (2009) Animal models of pulmonary arterial hypertension: the hope for etiological discovery and pharmacological cure. Am J Physiol Lung Cell Mol Physiol 297:L1013–L1032CrossRefGoogle Scholar
  38. 38.
    Sakuma F, Miyata M, Kasukawa R (1999) Suppressive effect of prostaglandin E1 on pulmonary hypertension induced by monocrotaline in rats. Lung 177:77–88CrossRefGoogle Scholar
  39. 39.
    Cowan KN, Heilbut A, Humpl T et al (2000) Complete reversal of fatal pulmonary hypertension in rats by a serine elastase inhibitor. Nat Med 6:698–702CrossRefGoogle Scholar
  40. 40.
    Kwon JH, Kim KC, Cho MS et al (2013) An inhibitory effect of tumor necrosis factor-alpha antagonist to gene expression in monocrotaline-induced pulmonary hypertensive rats model Korean. J Pediatr 56:116–124Google Scholar
  41. 41.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408CrossRefGoogle Scholar
  42. 42.
    Sasaki Y, Suzuki H, Itoh S et al (2012) K-134, a phosphodiesterase 3 inhibitor, improves gait disturbance and hindlimb blood flow impairment in rat peripheral artery disease models. Eur J Pharmacol 689:132–138CrossRefGoogle Scholar
  43. 43.
    Okumura K, Kato H, Honjo O et al (2015) Carvedilol improves biventricular fibrosis and function in experimental pulmonary hypertension. J Mol Med (Berl) 93:663–674CrossRefGoogle Scholar
  44. 44.
    Comeglio P, Morelli A, Adorini L et al (2017) Beneficial effects of bile acid receptor agonists in pulmonary disease models. Expert Opin Investig Drugs 26:1215–1228CrossRefGoogle Scholar
  45. 45.
    Comeglio P, Filippi S, Sarchielli E et al (2018) Therapeutic effects of obeticholic acid (OCA) treatment in a bleomycin-induced pulmonary fibrosis rat model. J Endocrinol Invest. 2018. (epub ahead of print) CrossRefPubMedGoogle Scholar
  46. 46.
    Sztuka K, Jasińska-Stroschein M (2017) Animal models of pulmonary arterial hypertension: a systematic review and meta-analysis of data from 6126 animals. Pharmacol Res 125:201–214CrossRefGoogle Scholar
  47. 47.
    Rubin LJ, Badesch DB, Barst RJ et al (2002) Bosentan therapy for pulmonary arterial hypertension. N Engl J Med 346:896–903CrossRefGoogle Scholar
  48. 48.
    Galiè N, Brundage BH, Ghofrani HA et al (2009) Tadalafil therapy for pulmonary arterial hypertension. Circulation 119:2894–2903CrossRefGoogle Scholar
  49. 49.
    Ranchoux B, Antigny F, Rucker-Martin C et al (2015) Endothelial-to-mesenchymal transition in pulmonary hypertension. Circulation 131:1006–1018CrossRefGoogle Scholar
  50. 50.
    Yang J, Li X, Al-Lamki RS et al (2010) Smad-dependent and smad-independent induction of id1 by prostacyclin analogues inhibits proliferation of pulmonary artery smooth muscle cells in vitro and in vivo. Circ Res 107:252–262CrossRefGoogle Scholar
  51. 51.
    Hashimoto N, Phan SH, Imaizumi K et al (2010) Endothelial-mesenchymal transition in bleomycin-induced pulmonary fibrosis. Am J Respir Cell Mol Biol 43:161–172CrossRefGoogle Scholar
  52. 52.
    Ahmedat AS, Warnken M, Seemann WK et al (2013) Pro-fibrotic processes in human lung fibroblasts are driven by an autocrine/paracrine endothelinergic system. Br J Pharmacol 168:471–487CrossRefGoogle Scholar
  53. 53.
    Wermuth PJ, Li Z, Mendoza FA, Jimenez SA (2016) Stimulation of transforming growth factor-β1-induced endothelial-to-mesenchymal transition and tissue fibrosis by endothelin-1 (ET-1): a novel profibrotic effect of ET-1. PLoS One 11:e0161988CrossRefGoogle Scholar
  54. 54.
    Breier G, Risau W (1996) The role of vascular endothelial growth factor in blood vessel formation. Trends Cell Biol 6:454–456CrossRefGoogle Scholar
  55. 55.
    Kelland NF, Kuc RE, McLean DL et al (2010) Endothelial cell-specific ETB receptor knockout: autoradiographic and histological characterisation and crucial role in the clearance of endothelin-1. Can J Physiol Pharmacol 88(6):644–651CrossRefGoogle Scholar
  56. 56.
    Leask A (2010) Potential therapeutic targets for cardiac fibrosis: TGFbeta, angiotensin, endothelin, CCN2, and PDGF, partners in fibroblast activation. Circ Res 106(11):1675–1680CrossRefGoogle Scholar
  57. 57.
    Swigris JJ, Brown KK (2010) The role of endothelin-1 in the pathogenesis of idiopathic pulmonary fibrosis. BioDrugs 24(1):49–54CrossRefGoogle Scholar
  58. 58.
    Rosenzweig BL, Imamura T, Okadome T et al (1995) Cloning and characterization of a human type II receptor for bone morphogenetic proteins. Proc Natl Acad Sci USA 92:7632–7636CrossRefGoogle Scholar
  59. 59.
    Du L, Sullivan CC, Chu D et al (2003) Signaling molecules in nonfamilial pulmonary hypertension. N Engl J Med 348:500–509CrossRefGoogle Scholar
  60. 60.
    Dewachter L, Adnot S, Guignabert C et al (2009) Bone morphogenetic protein signalling in heritable versus idiopathic pulmonary hypertension. Eur Respir J 34:1100–1110CrossRefGoogle Scholar
  61. 61.
    Thomson JR, Machado RD, Pauciulo MW et al (2000) Sporadic primary pulmonary hypertension is associated with germline mutations of the gene encoding BMPR-II, a receptor member of the TGF-beta family. J Med Genet 37(10):741–745CrossRefGoogle Scholar
  62. 62.
    Atkinson C, Stewart S, Upton PD et al (2002) Primary pulmonary hypertension is associated with reduced pulmonary vascular expression of type II bone morphogenetic protein receptor. Circulation 105:1672–1678CrossRefGoogle Scholar
  63. 63.
    Rouillard AD, Gundersen GW, Fernandez NF et al (2016) The harmonizome: a collection of processed datasets gathered to serve and mine knowledge about genes and proteins. Database (Oxford) 2016:1–16CrossRefGoogle Scholar
  64. 64.
    Shenoy V, Qi Y, Katovich MJ, Raizada MK (2011) ACE2, a promising therapeutic target for pulmonary hypertension. Curr Opin Pharmacol 11(2):150–155CrossRefGoogle Scholar
  65. 65.
    Dai HL, Guo Y, Guang XF et al (2013) The changes of serum angiotensin-converting enzyme 2 in patients with pulmonary arterial hypertension due to congenital heart disease. Cardiology 124:208–212CrossRefGoogle Scholar
  66. 66.
    Ferreira AJ, Shenoy V, Yamazato Y et al (2009) Evidence for angiotensin-converting enzyme 2 as a therapeutic target for the prevention of pulmonary hypertension. Am J Respir Crit Care Med 179(11):1048–1054CrossRefGoogle Scholar
  67. 67.
    Jiang F, Yang J, Zhang Y et al (2014) Angiotensin-converting enzyme 2 and angiotensin 1–7: novel therapeutic targets. Nat Rev Cardiol 11:413–426CrossRefGoogle Scholar
  68. 68.
    Yang J, Li X, Al-Lamki RS et al (2013) Sildenafil potentiates bone morphogenetic protein signaling in pulmonary arterial smooth muscle cells and in experimental pulmonary hypertension. Arterioscler Thromb Vasc Biol 33(1):34–42CrossRefGoogle Scholar
  69. 69.
    Thompson AAR, Lawrie A (2017) Targeting vascular remodeling to treat pulmonary arterial hypertension. Trends Mol Med 23(1):31–45CrossRefGoogle Scholar
  70. 70.
    Schermul RT, Kreisselmeier KP, Ghofrani HA et al (2004) Chronic sildenafil treatment inhibits monocrotaline-induced pulmonary hypertension in rats. Am J Respir Crit Care Med 169:39–45CrossRefGoogle Scholar
  71. 71.
    Sawamura F, Kato M, Fujita K et al (2009) Tadalafil, a long-acting inhibitor of PDE5, improves pulmonary hemodynamics and survival rate of monocrotaline-induced pulmonary artery hypertension in rats. J Pharmacol Sci 111:235–243CrossRefGoogle Scholar
  72. 72.
    Lee DS, Kim YK, Jung YW (2010) Simvastatin, sildenafil and their combination in monocrotaline induced pulmonary arterial hypertension. Korean Circ J 40:659–664CrossRefGoogle Scholar
  73. 73.
    Yen CH, Leu S, Lin YC et al (2010) Sildenafil limits monocrotaline-induced pulmonary hypertension in rats through suppression of pulmonary vascular remodeling. J Cardiovasc Pharmacol 55:574–584CrossRefGoogle Scholar
  74. 74.
    Arif SA, Poon H (2011) Tadalafil: a long-acting phosphodiesterase-5 inhibitor for the treatment of pulmonary arterial hypertension. Clin Ther 33:993–1004CrossRefGoogle Scholar
  75. 75.
    Schroll S, Sebah D, Wagner M et al (2013) Improvement of exercise capacity in monocrotaline-induced pulmonary hypertension by the phosphodiesterase-5 inhibitor Vardenafil. Respir Physiol Neurobiol 186:61–64CrossRefGoogle Scholar

Copyright information

© Italian Society of Endocrinology (SIE) 2019

Authors and Affiliations

  • P. Comeglio
    • 1
  • S. Filippi
    • 2
  • E. Sarchielli
    • 3
  • A. Morelli
    • 3
  • I. Cellai
    • 1
  • C. Corno
    • 1
  • L. Adorini
    • 4
  • G. B. Vannelli
    • 3
  • M. Maggi
    • 1
    • 5
  • L. Vignozzi
    • 1
    • 5
    Email author
  1. 1.Sexual Medicine and Andrology Unit, Department of Biomedical, Experimental and Clinical SciencesUniversity of Florence, AOU CareggiFlorenceItaly
  2. 2.Interdepartmental Laboratory of Functional and Cellular Pharmacology of Reproduction, Department of NEUROFARBAUniversity of FlorenceFlorenceItaly
  3. 3.Department of Experimental and Clinical MedicineUniversity of FlorenceFlorenceItaly
  4. 4.Intercept PharmaceuticalsNew YorkUSA
  5. 5.I.N.B.B. (Istituto Nazionale Biostrutture E Biosistemi)RomeItaly

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