Abstract
Mitochondrial dysfunction is a common mediator of disease and organ injury. Although recent studies show that inducing mitochondrial biogenesis (MB) stimulates cell repair and regeneration, only a limited number of chemicals are known to induce MB. To examine the impact of the β-adrenoceptor (β-AR) signaling pathway on MB, primary renal proximal tubule cells (RPTC) and adult feline cardiomyocytes were exposed for 24 h to multiple β-AR agonists: isoproterenol (nonselective β-AR agonist), (±)-(R*,R*)-[4-[2-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]propyl]phenoxy] acetic acid sodium hydrate (BRL 37344) (selective β3-AR agonist), and formoterol (selective β2-AR agonist). The Seahorse Biosciences (North Billerica, MA) extracellular flux analyzer was used to quantify carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP)-uncoupled oxygen consumption rate (OCR), a marker of maximal electron transport chain activity. Isoproterenol and BRL 37244 did not alter mitochondrial respiration at any of the concentrations examined. Formoterol exposure resulted in increases in both FCCP-uncoupled OCR and mitochondrial DNA (mtDNA) copy number. The effect of formoterol on OCR in RPTC was inhibited by the β-AR antagonist propranolol and the β2-AR inverse agonist 3-(isopropylamino)-1-[(7-methyl-4-indanyl)oxy]butan-2-ol hydrochloride (ICI-118,551). Mice exposed to formoterol for 24 or 72 h exhibited increases in kidney and heart mtDNA copy number, peroxisome proliferator-activated receptor γ coactivator 1α, and multiple genes involved in the mitochondrial electron transport chain (F0 subunit 6 of transmembrane F-type ATP synthase, NADH dehydrogenase subunit 1, NADH dehydrogenase subunit 6, and NADH dehydrogenase [ubiquinone] 1β subcomplex subunit 8). Cheminformatic modeling, virtual chemical library screening, and experimental validation identified nisoxetine from the Sigma Library of Pharmacologically Active Compounds and two compounds from the ChemBridge DIVERSet that increased mitochondrial respiratory capacity. These data provide compelling evidence for the use and development of β2-AR ligands for therapeutic MB.
Footnotes
L.P.W. is funded by the National Institutes of Health National Cancer Institute [Grant T32 CA119945-04] and the National Institutes of Health National Institute of Environmental Health Sciences [Grant F32 ES020103-01]. R.G.S. is funded by the National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grants DK062028, DK071997], the National Institutes of Health National Institute of Environmental Health Sciences [Grant ES012878], the National Institutes of Health National Institute of General Medical Sciences [Grant GM084147], and the Biomedical Laboratory Research and Development Program of the Department of Veterans Affairs. Animal facilities were funded by the National Institutes of Health National Center for Research Resources [Grant C06 RR-015455].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
ABBREVIATIONS:
- MB
- mitochondrial biogenesis
- AFC
- adult feline cardiomyocytes
- ATP6
- F0 subunit 6 of transmembrane F-type ATP synthase
- β-AR
- β-adrenoceptor
- CB
- ChemBridge
- Ct
- threshold cycle
- DMSO
- dimethyl sulfoxide
- FCCP
- carbonylcyanide p-trifluoromethoxyphenylhydrazone
- HPLC
- high-performance liquid chromatography
- LOPAC
- Library of Pharmacologically Active Compounds
- mtDNA
- mitochondrial DNA
- ND1
- NADH dehydrogenase subunit 1
- ND6
- NADH dehydrogenase subunit 6
- NDUFB8
- NADH dehydrogenase [ubiquinone] 1β subcomplex subunit 8
- OCR
- oxygen consumption rate
- PCR
- polymerase chain reaction
- PGC-1α
- peroxisome proliferator-activated receptor γ coactivator 1-α
- RMSD
- root mean square deviation
- RPTC
- renal proximal tubule cells
- SRT1720
- N-[2-[3-(piperazin-1-ylmethyl)imidazo[2,1-b][1,3]thiazol-6-yl]phenyl]quinoxaline-2-carboxamide
- TC
- Tanimoto Coefficient
- BRL 37344
- (±)-(R*,R*)-[4-[2-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]propyl]phenoxy] acetic acid sodium hydrate
- ICI-118,551
- 3-(isopropylamino)-1-[(7-methyl-4-indanyl)oxy]butan-2-ol hydrochloride.
- Received January 5, 2012.
- Accepted April 5, 2012.
- U.S. Government work not protected by U.S. copyright
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