Figure 4.

Liposome containing short-chain ceramide C6 treatment in MCD diet increases AMPK activation/phosphorylation and enhances the endogenous anti-oxidative stress signaling pathway without inducing apoptosis. (A) Representative Western blot analysis demonstrated an upregulation in activation/phosphorylation of AMPK by Lip-C6 treatment in MCD-fed mice in comparison with MCD-fed mice (n = 2 for each condition). (B) Inter-variability was assessed by protein analysis of pooled samples of each condition and densitometry scanning. MCD-fed mice have reduced AMPK protein levels in comparison with control diet-fed mice. Absolute levels of phosphorylated AMPK (P-AMPK) were highly induced in MCD-fed mice treated with Lip-C6 relative to MCD-fed mice and MCD-fed Lip-G treated mice. (C) Representative Western blot analysis demonstrated that Keap-1 protein expression was absent in MCD-fed mice, whereas Nrf2 and NQO1 protein expression were upregulated in MCD-fed mice in comparison with CD-fed mice (n = 2 for each condition). (D) Lip-C6 treatment in MCD-fed mice did not induce apoptosis by JNK activation/phosphorylation. (E) Cleaved poly (ADP-ribose) polymerase (PARP) and cleaved Caspase 3 protein expression showed to be absent in MCD-fed mice with no changes observed in Lip-C6 treated mice (n = 2 for each condition).