Fig. 5.

2-Hydroxyglutarate (2HG) activates adenosine 5’-monophosphate (AMP)-activated protein kinase (AMPK) in microglial cells. a BV-2 cells were incubated with lipopolysaccharide (LPS) (100 ng/mL) for the indicated times after a 30 min 2HG (1 mM) pretreatment. The ATP/ADP ratio was measured as described in the metabolic assays section. *P < 0.05 and **P < 0.01 compared to Con. ^#P < 0.05 compared to LPS. b Cells were stimulated with LPS or 2HG, and the LPS-induced initial oxygen consumption rate (OCR) was recorded every 10 min in the XF-24 analyzer. *P < 0.05 and **P < 0.01 compared to Con. ^#P < 0.05 and ^##P < 0.01 compared to LPS. c, d. Cells were treated with LPS or 2HG, and the levels of the indicated phosphorylated (p-) and total proteins were analyzed using Western blotting. The levels of all proteins were quantified using densitometry and normalized to α-tubulin. *P < 0.05 compared to Con. ^#P < 0.05 compared to LPS. e Cells were pretreated with the AMPK antagonist Compound C (Com C), and the anti-inflammatory effect of 2HG was partially reversed. **P < 0.01 compared to LPS. ^#P < 0.05 and ^##P < 0.01 compared to 2HG + LPS. All results are presented as the means ± SD (n = 3) and are representative of three independent experiments