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Effects of Fatty Acids on CYP2A5 and Nrf2 Expression in Mouse Primary Hepatocytes

机译:脂肪酸对小鼠原代肝细胞CYP2A5和Nrf2表达的影响

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Abnormal fatty acid metabolism is observed throughout nonalcoholic fatty liver disease (NAFLD) pathogenesis, and fatty acid storage is an important inducing factor in insulin resistance, lipid oxidation, hepatic cell damage, and inflammation. During NAFLD pathogenesis, changes in blood and liver contents of different fatty acid types also vary. Cytochrome P450 2A5 (CYP2A5), an important enzyme in mouse liver, metabolizes many drugs and activates multiple pro-carcinogens with widely varying structures. According to the changes in liver fatty acid profiles observed in NAFLD animal models developed in our laboratory and others, saturated (PA/palmitic, and SA/stearic acids) and unsaturated (OA/oleic, LA/linoleic, ALA/alpha-linolenic and AA/arachidonic acids) fatty acids were selected to induce mouse primary hepatocytes, at concentrations under 1 mM, as detected by MTT assay. After 24 h treatment with various fatty acid concentrations and types, CYP2A5 mRNA and protein amounts, and enzyme activity were determined by real-time PCR, Western blot, and Coumarin 7-hydroxylation, respectively. Meanwhile, Nrf2 mRNA and protein levels were evaluated by real-time PCR and Western blot. The results indicated that saturated fatty acids are more potent in inducing CYP2A5 than unsaturated ones, except arachidonic acid. In addition, the changes in CYP2A5 expression were consistent with the alterations observed in Nrf2 expression, indicating that Nrf2 might play a regulatory role in CYP2A5 expression.
机译:在非酒精性脂肪肝疾病(NAFLD)的整个发病机理中均观察到异常的脂肪酸代谢,并且脂肪酸存储是胰岛素抵抗,脂质氧化,肝细胞损伤和炎症的重要诱导因素。在NAFLD发病机理中,不同脂肪酸类型的血液和肝脏含量变化也不同。细胞色素P450 2A5(CYP2A5)是小鼠肝脏中的一种重要酶,可代谢多种药物并激活多种具有多种结构的致癌物。根据在我们实验室和其他实验室开发的NAFLD动物模型中观察到的肝脏脂肪酸谱变化,饱和(PA /棕榈酸和SA /硬脂酸)和不饱和(OA /油酸,LA /亚油酸,ALA /α-亚麻酸和如通过MTT测定所检测的,选择浓度低于1mM的AA /花生四烯酸)脂肪酸以诱导小鼠原代肝细胞。用各种脂肪酸浓度和类型处理24小时后,分别通过实时PCR,Western印迹和香豆素7-羟基化测定CYP2A5 mRNA和蛋白质含量以及酶活性。同时,通过实时PCR和蛋白质印迹法评估Nrf2 mRNA和蛋白质水平。结果表明,除了花生四烯酸外,饱和脂肪酸比不饱和脂肪酸更有效地诱导CYP2A5。另外,CYP2A5表达的变化与在Nrf2表达中观察到的变化一致,表明Nrf2可能在CYP2A5表达中起调节作用。

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