PPARα regulates the expression of human arylacetamide deacetylase involved in drug hydrolysis and lipid metabolism
PPARα regulates the expression of human arylacetamide deacetylase involved in drug hydrolysis and lipid metabolism
复制标题
DOI：
10.1016/j.bcp.2022.115010
复制
发表时间：
2022-03-22
影响因子：
5.8
通讯作者：
Nakajima, Miki
中科院分区：
文献类型：
--
作者：
Morikawa, Tomomi;Fukami, Tatsuki;Nakajima, Miki
人芳基乙酰胺脱乙酰基酶(AADAC)能水解酮康唑、利福平等多种含乙酰基的药物。关于人AADAC在药物代谢中的作用的知识正在积累，但其表达的调控机制尚未阐明。在小鼠中，Aadac的表达可能受过氧化体增殖物激活受体α(PPARα)的调节。这项研究考察了人类AADAC是否受到PPARα的调节，PPARα广泛地调节脂质代谢相关基因的表达。人肝癌HUH-7细胞经非诺贝酸和WY-14643、PPARα配体处理后，AADAC基因和蛋白水平显著升高。PPARα表达下调和过表达分别导致AADAC表达降低和增加。荧光素酶分析表明，AADAC启动子区域-193/-181的直接重复序列1(DR1)负责PPARα的反式激活。染色质免疫沉淀分析显示PPARα与DR1结合。因此，证实了人AADAC是由PPARα通过与DR1结合来调节的。油红O染色显示，在Huh-7细胞中过表达AADAC抑制了游离脂肪酸处理后的脂质积聚。用AADAC抑制剂二异丙基氟磷酸盐处理后，抑制作用恢复。AADAC基因敲除可恢复WY-14643介导的抑制脂质堆积的作用。这些结果表明，AADAC具有抑制细胞脂质蓄积的作用。综上所述，本研究证实了PPARα对人AADAC的调节及其在脂质蓄积中的意义。
Human arylacetamide deacetylase (AADAC) hydrolyzes various drugs containing an acetyl group, such as ketoconazole and rifampicin. Knowledge about the role of human AADAC in drug metabolism is accumulating, but the regulatory mechanism of its expression has not been elucidated. In mice, it has been suggested that Aadac expression may be regulated by peroxisome proliferator-activated receptor alpha (Ppar alpha). This study examined whether human AADAC is regulated by PPAR alpha, which widely regulates the expression of lipid metabolismrelated genes. In human hepatoma Huh-7 cells, AADAC mRNA and protein levels were significantly increased by treatment with fenofibric acid and WY-14643, PPAR alpha ligands. Knockdown and overexpression of PPAR alpha resulted in decreased and increased expression of AADAC, respectively. Luciferase assays revealed that the direct repeat 1 (DR1) at -193/-181 in the AADAC promoter region is responsible for transactivation by PPAR alpha. Chromatin immunoprecipitation assays revealed the binding of PPAR alpha to DR1. Thus, it was demonstrated that human AADAC is regulated by PPAR alpha through binding to DR1. Oil red O staining showed that overexpression of AADAC in Huh-7 cells suppressed lipid accumulation after treatment with free fatty acids. The suppression was restored by treatment with diisopropyl fluorophosphate, an AADAC inhibitor. The WY-14643-mediated suppression of lipid accumulation was restored by AADAC knockdown. These results suggested that AADAC has a role in suppressing cellular lipid accumulation. In conclusion, this study demonstrated the regulation of human AADAC by PPAR alpha and its significance in lipid accumulation.