2017年4月
Intracellular Angiotensin-II Interacts With Nuclear Angiotensin Receptors in Cardiac Fibroblasts and Regulates RNA Synthesis, Cell Proliferation, and Collagen Secretion
JOURNAL OF THE AMERICAN HEART ASSOCIATION
- 巻
- 6
- 号
- 4
- 開始ページ
- pii: e004965
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1161/JAHA.116.004965
- 出版者・発行元
- WILEY
Background-Cardiac fibroblasts play important functional and pathophysiological roles. Intracellular ("intracrine") angiotensin-II (Ang-II) signaling regulates intercellular communication, excitability, and gene expression in cardiomyocytes; however, the existence and role of intracrine Ang-II signaling in cardiac fibroblasts is unstudied. Here, we evaluated the localization of Ang-II receptors on atrial fibroblast nuclei and associated intracrine effects of potential functional significance.
Methods and Results-Immunoblots of subcellular protein-fractions from isolated canine atrial fibroblasts indicated the presence of nuclear Ang-II type 1 receptors (AT1Rs) and Ang-II type 2 receptors (AT2Rs). Fluorescein isothiocyanate-Ang-II binding displaceable by AT1R- and AT2R-blockers was present on isolated fibroblast nuclei. G-protein subunits, including G alpha q/11, G alpha i/3, and G beta, were observed in purified fibroblast nuclear fractions by immunoblotting and intact-fibroblast nuclei by confocal immunocytofluorescence microscopy. Nuclear AT1Rs and AT2Rs regulated de novo RNA synthesis ([alpha P-32]UTP incorporation) via IP3R- and NO-dependent pathways, respectively. In intact cultured fibroblasts, intracellular Ang-II release by photolysis of a membrane-permeable caged Ang-II analog led to IP3R-dependent nucleoplasmic Ca2+-liberation, with IP3R3 being the predominant nuclear isoform. Intracellular Ang-II regulated fibroblast proliferation ([H-3] thymidine incorporation), collagen-1A1 mRNA-expression, and collagen secretion. Intracellular Ang-II and nuclear AT1R protein levels were significantly increased in a heart failure model in which atrial fibrosis underlies atrial fibrillation.
Conclusions-Fibroblast nuclei possess AT1R and AT2R binding sites that are coupled to intranuclear Ca2+-mobilization and NO liberation, respectively. Intracellular Ang-II signaling regulates fibroblast proliferation, collagen gene expression, and collagen secretion. Heart failure upregulates Ang-II intracrine signaling-components in atrial fibroblasts. These results show for the first time that nuclear angiotensin-II receptor activation and intracrine Ang-II signaling control fibroblast function and may have pathophysiological significance.
Methods and Results-Immunoblots of subcellular protein-fractions from isolated canine atrial fibroblasts indicated the presence of nuclear Ang-II type 1 receptors (AT1Rs) and Ang-II type 2 receptors (AT2Rs). Fluorescein isothiocyanate-Ang-II binding displaceable by AT1R- and AT2R-blockers was present on isolated fibroblast nuclei. G-protein subunits, including G alpha q/11, G alpha i/3, and G beta, were observed in purified fibroblast nuclear fractions by immunoblotting and intact-fibroblast nuclei by confocal immunocytofluorescence microscopy. Nuclear AT1Rs and AT2Rs regulated de novo RNA synthesis ([alpha P-32]UTP incorporation) via IP3R- and NO-dependent pathways, respectively. In intact cultured fibroblasts, intracellular Ang-II release by photolysis of a membrane-permeable caged Ang-II analog led to IP3R-dependent nucleoplasmic Ca2+-liberation, with IP3R3 being the predominant nuclear isoform. Intracellular Ang-II regulated fibroblast proliferation ([H-3] thymidine incorporation), collagen-1A1 mRNA-expression, and collagen secretion. Intracellular Ang-II and nuclear AT1R protein levels were significantly increased in a heart failure model in which atrial fibrosis underlies atrial fibrillation.
Conclusions-Fibroblast nuclei possess AT1R and AT2R binding sites that are coupled to intranuclear Ca2+-mobilization and NO liberation, respectively. Intracellular Ang-II signaling regulates fibroblast proliferation, collagen gene expression, and collagen secretion. Heart failure upregulates Ang-II intracrine signaling-components in atrial fibroblasts. These results show for the first time that nuclear angiotensin-II receptor activation and intracrine Ang-II signaling control fibroblast function and may have pathophysiological significance.
- リンク情報
- ID情報
-
- DOI : 10.1161/JAHA.116.004965
- ISSN : 2047-9980
- Web of Science ID : WOS:000404098500029