Short-chain C2 ceramide induces heme oxygenase-1 expression by upregulating AMPK and MAPK signaling pathways in rat primary astrocytes

• C2 ceramide has antioxidant and cytoprotective effects in rat primary astrocytes.
• C2 ceramide increases phase II antioxidant enzyme gene expression in astrocytes.
• C2 ceramide increases Nrf2/ARE pathway signaling in astrocytes.
• AMPK and MAPKs are involved in C2 ceramide-induced HO-1 expression via Nrf2/ARE.
• C2 ceramide has therapeutic potential for neurodegenerative diseases.

Ceramide belongs to the group of sphingolipid metabolites that are produced in the brain and peripheral systems and act as intracellular second messengers. Although some physiological roles of ceramide have been reported in the brain, the role of ceramide in astrocytes has not been clearly demonstrated. In the present study, we investigated the antioxidant effects of the cell-permeable short-chain C2 ceramide in rat brain astrocytes. C2 ceramide inhibited hydrogen peroxide-induced reactive oxygen species generation and subsequent cell death in rat primary astrocytes. C2 ceramide increased the expression of phase II antioxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinine oxidoreductase 1 (NQO1), and superoxide dismutase (SOD) that are under the control of Nrf2/ARE signaling pathways. Detailed mechanistic studies revealed that C2 ceramide increased the nuclear translocation and DNA binding of nuclear factor-E2-related factor 2 (Nrf2) and c-Jun to the antioxidant response element (ARE), and increased ARE-mediated transcriptional activity. Moreover, C2 ceramide increased the interaction between Nrf2 and c-Jun as shown by antibody co-immunoprecipitation assay. Further analysis of signaling pathways revealed that AMPK and MAP kinases are involved in HO-1 expression by modulating ARE-mediated transcriptional activity. Therefore, the upregulation of antioxidant enzymes by C2 ceramide may be a potential therapeutic modality for neurodegenerative diseases that are accompanied by oxidative stress.