miR‐29a‐3p promotes the regulatory role of eicosapentaenoic acid in the NLRP3 inflammasome and autophagy in microglial cells

Abstract

AbstractEicosapentaenoic acid (EPA) has been reported to play an anti‐inflammatory and antioxidative stress role in a series of human diseases, including major depressive disorder. However, its exact mechanism is still largely unknown. Mouse BV‐2 cells were treated with lipopolysaccharide (LPS) to induce an in vitro inflammatory cell model of depression. Cytotoxic effects were assessed with MTT and lactate dehydrigebase release assays. Cytokine mediators were elevated by western blot and enzyme‐linked immunosorbent assays. Autophagy‐relators were determined by immunofluorescence and western blot analyses. Interaction relationships among molecules were evaluated utilizing chromatin immunoprecipitation and dual luciferase assays. Methylated miR‐29a‐3p was detected via methylation‐specific polymerase chain reaction. EPA treatment at 60 μM had no cytotoxic effects on BV2 cells and significantly inhibited the LPS‐induced inflammatory response and NLRP3 inflammasome but activated autophagy, while all these effects were reversed by the autophagy inhibitor 3‐MA. Importantly, miR‐29a‐3p exhibited a role similar to that of EPA in LPS‐treated BV2 cells. Mechanistically, EPA treatment elevated miR‐29a‐3p by repressing its promoter methylation. MAPK8 was a direct target of miR‐29a‐3p. Inhibition of miR‐29a‐3p greatly diminished the regulatory roles mediated by EPA in LPS‐treated BV2 cells, while these roles were further impeded after MAPK8 silencing. To conclude, our data demonstrated that EPA treatment alleviated LPS‐induced NLRP3 inflammasomes by activating autophagy via regulation of miR‐29a‐3p/MAPK8 signaling, which further elucidates the potential antidepressant mechanism of EPA.