The β2-Adrenoceptor Agonist Formoterol Stimulates Mitochondrial Biogenesis
The β2-Adrenoceptor Agonist Formoterol Stimulates Mitochondrial Biogenesis
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DOI：
10.1124/jpet.112.191528
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发表时间：
2012-07
影响因子：
3.5
通讯作者：
Lauren P. Wills;Richard E Trager;G. Beeson;Christopher C. Lindsey;Y. Peterson;C. Beeson;R. Schnellmann
中科院分区：
文献类型：
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作者：
Lauren P. Wills;Richard E Trager;G. Beeson;Christopher C. Lindsey;Y. Peterson;C. Beeson;R. Schnellmann
线粒体功能障碍是疾病和器官损伤的常见介质。虽然最近的研究表明，诱导线粒体生物合成（MB）刺激细胞修复和再生，只有有限数量的化学物质是已知的诱导MB。为了检查β-肾上腺素受体（β-AR）信号传导通路对MB的影响，将原代肾近端小管细胞（RPTC）和成年猫心肌细胞暴露于多种β-AR激动剂24小时：异丙（非选择性β-AR激动剂），（±）-（R*，R*）-[4-[2-[[2-（3-氯苯基）-2-羟乙基]氨基]丙基]苯氧基]乙酸钠水合物（BRL 37344）（选择性β3-AR激动剂）和福莫特罗（选择性β2-AR激动剂）。Seahorse Biosciences（North Billerica，MA）细胞外通量分析仪用于定量羰基氰化物对三氟甲氧基苯腙（FCCP）-解偶联耗氧率（OCR），这是最大电子传递链活性的标志物。异丙肾上腺素和BRL 37244在任何检测浓度下均未改变线粒体呼吸。福莫特罗暴露导致FCCP-非偶联OCR和线粒体DNA（mtDNA）拷贝数增加。β-AR拮抗剂普萘洛尔和β2-AR反向激动剂3-（异丙基氨基）-1-[（7-甲基-4-茚满基）氧基]丁-2-醇盐酸盐（ICI-118，551）可抑制福莫特罗对RPTC中OCR的影响。暴露于福莫特罗24或72 h的小鼠表现出肾脏和心脏mtDNA拷贝数、过氧化物酶体增殖物激活受体γ共激活因子1α和参与线粒体电子传递链的多个基因（跨膜F型ATP合酶的F0亚基6、NADH脱氢酶亚基1、NADH脱氢酶亚基6和NADH脱氢酶[泛醌] 1β亚复合物亚基8）增加。化学信息学建模、虚拟化学文库筛选和实验验证从Sigma药理学活性化合物文库中鉴定出尼索西汀，并从ChemBridge DIVERSet中鉴定出两种化合物，其增加线粒体呼吸能力。这些数据为β2-AR配体用于治疗MB的使用和开发提供了令人信服的证据。
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.