Skip to main content
. 2014 Oct 20;21(12):1741–1758. doi: 10.1089/ars.2013.5587

FIG. 1.

FIG. 1.

H2S donors robustly enhanced AMPK activation in BV2 microglial cells. (A) Western blot analysis of ADT-OH effects on AMPK activation (phosphorylation) in BV2 cells without LPS stimulation at the indicated time points post ADT-OH treatment (n=3). Compared with nontreated control cells, ADT-OH-treated BV2 cells displayed significantly enhanced AMPK activation, as indicated by increased ratios of phosphorylated AMPK (p-AMPK)/AMPK (*p<0.05; **p<0.01). (B–D) Western blot analysis of effects of H2S donors ADT-OH (B), NaHS (C), and GYY4137 (D) on AMPK activation (phosphorylation) in BV2 cells in the presence of LPS stimulation (n=3). Compared with control cells without LPS and H2S donor treatment, LPS-treated cells displayed a significant decrease in AMPK activation for approximately 4 h post LPS stimulation (*p<0.05 or **p<0.01). ADT-OH (B), NaHS (C), and GYY4137 (D) significantly enhanced AMPK phosphorylation in LPS-stimulated BV2 cells. ##p<0.01, compared with cells treated with LPS alone (LPS) at the indicated time points. (E) Western blot analysis of ADT-OH effects on phosphorylation of ACC, a substrate of activated AMPK, in BV2 cells (n=4). Phosphorylation of ACC (p-ACC) was assessed to indicate AMPK activity. In the absence of LPS, compared with vehicle (Con), ADT-OH enhanced the ratios of p-ACC/ACC (*p<0.05). ADT-OH also remarkably enhanced p-ACC/ACC ratios in LPS-stimulated cells. ##p<0.01, compared with cells treated with LPS alone (LPS). ADT-OH enhancement of p-ACC/ACC ratios was significantly attenuated by the AMPK inhibitor compound C (CC) at 2.5 μM. $p<0.05, compared with cells treated with LPS and ADT-OH (LPS+ADT-OH). ACC, acetyl-CoA carboxylase; ADT-OH, 5-(4-hydroxyphenyl)-3H-1,2-dithiocyclopentene-3-thione; AMPK, AMP-activated protein kinase; GYY4137, (p-methoxyphenyl) morpholino-phosphinodithioic acid; H2S, hydrogen sulfide; LPS, lipopolysaccharide; NaHS, sodium hydrosulfide.