title: The protective effect of ursodeoxycholic acid in an in vitro model of the human fetal heart occurs via targeting cardiac fibroblasts.
abstract: Bile acids are elevated in the blood of women with intrahepatic cholestasis of pregnancy (ICP) and this may lead to fetal arrhythmia, fetal hypoxia and potentially fetal death in utero. The bile acid taurocholic acid (TC) causes abnormal calcium dynamics and contraction in neonatal rat cardiomyocytes. Ursodeoxycholic acid (UDCA), a drug clinically used to treat ICP, prevents adverse effects of TC. During development, the fetus is in a state of relative hypoxia. Although this is essential for the development of the heart and vasculature, resident fibroblasts can transiently differentiate into myofibroblasts and form gap junctions with cardiomyocytes in vitro, resulting in cardiomyocyte depolarization. We expanded on previously published work using an in vitro hypoxia model to investigate the differentiation of human fetal fibroblasts into myofibroblasts. Recent evidence shows that potassium channels are involved in maintaining the membrane potential of ventricular fibroblasts and that ATP-dependent potassium (KATP) channel subunits are expressed in cultured fibroblasts. KATP channels are a valuable target as they are thought to have a cardioprotective role during ischaemic and hypoxic conditions. We investigated whether UDCA could modulate fibroblast membrane potential. We established the isolation and culture of human fetal cardiomyocytes and fibroblasts to investigate the effect of hypoxia, TC and UDCA on human fetal cardiac cells. UDCA hyperpolarized myofibroblasts and prevented TC-induced depolarisation, possibly through the activation of KATP channels that are expressed in cultured fibroblasts. Also, similar to the rat model, UDCA can counteract TC-induced calcium abnormalities in human fetal cultures of cardiomyocytes and myofibroblasts. Under normoxic conditions, we found a higher number of myofibroblasts in cultures derived from human fetal hearts compared to cells isolated from neonatal rat hearts, indicating a possible increased number of myofibroblasts in human fetal hearts. Hypoxia further increased the number of human fetal and rat neonatal myofibroblasts. However, chronically administered UDCA reduced the number of myofibroblasts and prevented hypoxia-induced depolarisation. In conclusion, our results show that the protective effect of UDCA involves both the reduction of fibroblast differentiation into myofibroblasts, and hyperpolarisation of myofibroblasts, most likely through the stimulation of potassium channels, i.e. KATP channels. This could be important in validating UDCA as an antifibrotic and antiarrhythmic drug for treatment of failing hearts and fetal arrhythmia.
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK; Institute of Reproductive and Developmental Biology, Imperial College London, London, UK.
authors.forename: Francisca
authors.initials: F
authors.lastname: Schultz
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK.
authors.forename: Alveera
authors.initials: A
authors.lastname: Hasan
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK.
authors.forename: Anita
authors.initials: A
authors.lastname: Alvarez-Laviada
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK; Humanitas Clinical and Research Institute, Rozzano, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Miragoli
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK.
authors.forename: Navneet
authors.initials: N
authors.lastname: Bhogal
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Wells
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK.
authors.forename: Claire
authors.initials: C
authors.lastname: Poulet
authors.affiliation: Institute of Reproductive and Developmental Biology, Imperial College London, London, UK; Women's Health Academic Centre, King's College London, London, United Kingdom.
authors.forename: Jenny
authors.initials: J
authors.lastname: Chambers
authors.affiliation: Institute of Reproductive and Developmental Biology, Imperial College London, London, UK; Women's Health Academic Centre, King's College London, London, United Kingdom.
authors.forename: Catherine
authors.initials: C
authors.lastname: Williamson
authors.affiliation: National Heart and Lung Institute, Imperial Centre for Translational and Experimental Medicine, Imperial College London, London, UK. Electronic address: j.gorelik@imperial.ac.uk.
authors.forename: Julia
authors.initials: J
authors.lastname: Gorelik
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D015687:Cell Hypoxia / Q000187:drug effects; D002469:Cell Separation; D019610:Cytoprotection / Q000187:drug effects; D005318:Fetal Heart / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013656:Taurocholic Acid / Q000494:pharmacology; D014580:Ursodeoxycholic Acid / Q000494:pharmacology*
chemical_list: D013656:Taurocholic Acid; D014580:Ursodeoxycholic Acid; D002118:Calcium
keywords: ATP-Dependent potassium channel; Bile acids; Fetal arrhythmia; Fetal heart; Fibroblasts; Ursodeoxycholic acid