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

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Abstract

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/α-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.

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Abbreviations

Nrf2:

Nuclear factor erythroid 2-like 2

P450:

Cytochrome P450

NAFLD:

Nonalcoholic fatty liver disease

FAs:

Fatty acids

PA:

Palmitic acid

SA:

Stearic acid

OA:

Oleic acid

LA:

Linoleic acid

ALA:

α-linolenic acid

AA:

Arachidonic acid

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Acknowledgments

We thank Dr H. Raunio (University of Kuopio, Kuopio, Finland) for generously providing antibodies against Cyp2a5. This work was supported by the Natural Science Foundation of China, Grant No. 31172369 and the Natural Science Foundation of Heilongjiang Province in China, Grant No.C201444.

Author Contributions

All authors participated in the design, interpretation of the studies, and analysis of the data and review of the manuscript; CYZ, WQJ, YX, and ZXY conducted the experiments, CYZ and WQJ wrote the manuscript, and ZXY contributed.

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    Authors and Affiliations

    1. College of Veterinary Medicine, Northeast Agricultural University, No.59 Mucai Street, Xiangfang District, Harbin, 150030, Heilongjiang, People’s Republic of China

      Yizhe Cui, Xing Yi & Xiuying Zhang

    2. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, No. 2 Xinyang Road, Sartu District, Daqing, 163319, Heilongjiang, People’s Republic of China

      Yizhe Cui & Qiuju Wang

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    1. Yizhe Cui
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    Correspondence to Xiuying Zhang.

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    Yizhe Cui and Qiuju Wang are contributed equally to this work.

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    Cui, Y., Wang, Q., Yi, X. et al. Effects of Fatty Acids on CYP2A5 and Nrf2 Expression in Mouse Primary Hepatocytes. Biochem Genet 54, 29–40 (2016). https://doi.org/10.1007/s10528-015-9697-6

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