Valproic Acid Is a Novel Activator of AMP-Activated Protein Kinase and Decreases Liver Mass, Hepatic Fat Accumulation, and Serum Glucose in Obese Mice
Valproic Acid Is a Novel Activator of AMP-Activated Protein Kinase and Decreases Liver Mass, Hepatic Fat Accumulation, and Serum Glucose in Obese Mice
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DOI：
10.1124/mol.113.089755
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发表时间：
2014-01-01
影响因子：
3.6
通讯作者：
Bumpus, Namandje N.
中科院分区：
文献类型：
--
作者：
Avery, Lindsay B.;Bumpus, Namandje N.
丙戊酸（VPA）是一种广泛用于治疗癫痫的抗惊厥药。在这里，我们证明了丙戊酸钠是一种新的激活剂的AMP激活的蛋白激酶（AMPK），细胞代谢的关键调节，使用原代小鼠和人类肝细胞。用VPA孵育原代小鼠肝细胞导致磷酸化AMPK和乙酰辅酶A羧化酶（ACC）水平升高。使用原代人肝细胞重现了这一发现。用AMPK的小分子抑制剂化合物C（6-[4-（2-哌啶-1-基乙氧基）苯基]-3-吡啶-4-基吡唑并[1，5-a]嘧啶）预处理小鼠肝细胞，消除了用VPA处理后ACC的磷酸化。细胞色素P450抑制剂1-氨基苯并三唑阻断VPA刺激的AMPK磷酸化，表明VPA生物转化的要求。与此一致，与VPA相比，用VPA代谢物处理肝细胞导致AMPK/ACC磷酸化增加。用VPA处理ob/ob小鼠14天导致肝脏质量、肝脏脂肪蓄积和血清葡萄糖降低。这些结果与二甲双胍给药小鼠中观察到的结果一致。此外，基于靶向质谱的代谢组学试验显示，与溶剂处理小鼠相比，VPA处理ob/ob小鼠血清中的几种小分子丰度存在差异。这些研究首次确立了VPA及其代谢产物作为AMPK的体外激活剂。
Valproic acid (VPA) is a widely prescribed anticonvulsant for the treatment of epilepsy. Here we demonstrate that VPA is a novel activator of AMP-activated protein kinase (AMPK), a key regulator of cellular metabolism, using primary mouse and human hepatocytes. Incubation of primary mouse hepatocytes with VPA resulted in increased levels of phosphorylated AMPK and acetylCoA carboxylase (ACC). This finding was recapitulated using primary human hepatocytes. Pretreatment of mouse hepatocytes with a small-molecule inhibitor of AMPK, Compound C (6-[4-(2-piperidin-1-ylethoxy)phenyl]-3-pyridin-4-ylpyrazolo[1,5-a]pyrimidine), abrogated the phosphorylation of ACC following treatment with VPA. The cytochrome P450 inhibitor 1-aminobenzotriazole blocked the VPA-stimulated phosphorylation of AMPK, suggesting a requirement for biotransformation of VPA. In line with this, treatment of hepatocytes with metabolites of VPA resulted in increased phosphorylation of AMPK/ACC as compared with VPA. Treatment of ob/ob mice with VPA for 14 days resulted in decreased liver masses, hepatic fat accumulation, and serum glucose. These results paralleled those observed in mice treated with metformin. In addition, a targeted mass spectrometry-based metabolomics assay revealed several small molecules that were differentially abundant in the serum of ob/obmice treated with VPA as compared with vehicle-treated mice. These studies are the first to establish VPA and its metabolites as in vitro activators of AMPK.