Rho-Kinase Inhibition Ameliorates Metabolic Disorders through Activation of AMPK Pathway in Mice
Figure 6
Fasudil Activates AMPK in Vitro via LKB1 Pathway.
(A) In C2C12 myotubes, hydroxyfasudil (30 µmol/L) increased AMPK phosphorylation with a peak at 6 h after incubation (n = 3 each). (B) In C2C12 myotubes, hydroxyfasudil increased AMPK phosphorylation in a concentration-dependent manner after 6 hours incubation (n = 3 each). (C) Hydroxyfasudil (10 µmol/L for 48 h) significantly up-regulated mRNA expression of Cpt1b and Cox4il as well as NAD+/NADH ratio, all of which were inhibited by compound C (50 µmol/L) (n = 6 each). Results are normalized by the expression of Gapdh. (D) AMPK activation by hydroxyfasudil was significantly inhibited by siRNA for LKB1 and there was no additional effect by TAK1 or STO-609 inhibition (n = 3 each). Results are expressed as mean ± SEM. *P<0.05 vs. 0 h, †P<0.05 vs. 0 µM, ‡P<0.05 vs. 1 µM, §P<0.05 vs. fasudil without compound C, ¶P<0.05 vs. without hydroxyfasudil, siLKB1, siTAK1 and STO-609, #P<0.05 vs. hydroxyfasudil.