miR-146a alleviates the apoptosis of hippocampal neurons induced by microglia activation via targeting TRAF6

Objective: To identify the role of miR-146a and tumor necrosis factor receptor-associated factor 6 (TRAF6) for improving the apoptosis of hippocampal neurons induced by microglia activation. Methods: Mouse microglial cell line (BV2 cell) was employed and treated with lipopolysaccharide. Mouse hippocampal nerve cell line (HT22 cell) was then grown in BV2 conditioned medium, and miR-146a overexpression and silencing cell lines were constructed. CCK8 and clone formation test were utilized to evaluate the proliferation ability of the transfected cells, and the level of inflammatory factors was measured by ELISA. Apoptosis was determined extensively by flow cytometry. The apoptosis-related protein and TRAF6 protein expressions were verified by Western blot. TRAF6 was identified to be the target gene of miR-146a based on double Luciferase Report. Finally, both TRAF6 and miR-146a were used to treat HT22 cells and the above indexes were detected repeatedly. Results: Interleukin (IL)-1β, tumor necrosis factor-α, and IL-6 expressions in BV2 cells increased significantly. miR-146a overexpression distinctly increased the cell proliferation ability and B-cell lymphoma-2 expression ((Bcl-2, p < 0.05); meanwhile, the apoptosis rate of cells, apoptosis-related proteins (Bcl-2 associated X and cleaved caspase-3), and TRAF6 gene and protein expressions were significantly decreased ( p < 0.05). However, these above results were reversed for miR-146a silence. There is a targeting relationship between miR-146a and TRAF6. Silencing TRAF6 gene can promote HT22 cells’ proliferation and inhibit apoptosis. The effect of miR-146a on HT22 cells was reversed by adding TRAF6 mimics to miR-146a overexpression cells. Conclusion: miR-146a can inhibit the apoptosis of hippocampal neurons caused by microglia activation via targeting TRAF6 and down-regulating its expression.