Slc7a11 stimulates glutathione synthesis to preserve fatty acid metabolism in primary hepatocytes

ABSTRACT

Primary hepatocytes are widely used as a tool for studying metabolic function and regulation in the liver. However, the metabolic properties of primary hepatocytes are gradually lost after isolation. Here, we illustrated that fatty acid metabolism is the major compromised metabolic process in isolated primary hepatocytes, along with drastically decreased GSH and ROS content, while lipid peroxidation is increased. Gain- and loss-of-function studies revealed that Slc7a11 expression is critical in maintaining fatty acid metabolism and facilitating hormone-induced fatty acid metabolic events, which is synergistic with dexamethasone treatment. Intriguingly, Slc7a11 expression and dexamethasone treatment cooperatively upregulated AKT and AMPK signaling and mitochondrial complex expression in primary hepatocytes. Furthermore, direct treatment with reduced GSH or inhibition of ferroptosis is sufficient to drive protective effects on fatty acid metabolism in primary hepatocytes. Our results demonstrate that Slc7a11 expression in isolated primary hepatocytes induces GSH production, which protects against ferroptosis, to increase fatty acid metabolic gene expression, AKT and AMPK signaling and mitochondrial function in synergy with dexamethasone treatment, thereby efficiently preserving primary hepatocyte metabolic signatures, thus providing a promising approach to better reserve primary hepatocyte metabolic activities after isolation to potentially improve the understanding of liver biological functions from studies using primary hepatocytes.

KEYWORDS:

• Slc7a11
• GSH
• ferroptosis
• primary hepatocytes
• fatty acid metabolism
• dexamethasone
• AKT/AMPK signaling‌
• mitochondrial homeostasis

Acknowledgments

We thank the members of the Zhao lab for their delicate guidance and advice on this study.

Author contributions

Y.L., Y.F., K.W., W.L. and X.Y.Z. conceived the project and designed the research. Y.L., K.W. and Y.F. performed the studies. Y.L. W.L. and X.Y.Z. analyzed the data and wrote the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The authors confirm that the datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

Additional information

Funding

This work was funded by the National Key R&D Program of China (2020YFA0803603 to X.Y.Z), the National Natural Science Foundation of China (82070894 to X.Y. Z), Science and Technology Commission of Shanghai Municipality (21ZR1436500, 22ZR1479800 to X.Y.Z), Innovative research team of high-level local universities in Shanghai (SHSMU-ZDCX20212501 to X.Y.Z), Shanghai Frontiers Science Center of Cellular Homeostasis and Human Diseases.