Fig. 7

AMPK inhibition blunted rhFGF21-induced attenuation of inflammation and oxidative stress in PA-treated HK-2 cells. Phosphorylated AMPK was inhibited using Compound C (a ATP competitive AMPK inhibitor, 10 μM) in human renal tubular epithelial cells (HK-2), the cells were then pretreated with rhFGF21 (50 ng/ml) for 2 h and subsequently stimulated with overdoes palmitate acid (PA, 1 mM) for 24 h. a Western blot analysis showed the effects of rhFGF21 and Compound C pretreatment on PA-induced AMPK phosphorylation in HK-2 cells. Image J was used to quantify the ratio of p-AMPK/AMPK. GAPDH was used as the internal control. b and c The nuclear translocation levels of NF-κB p65 and Nrf2 in nuclear fractions were detected by Western blot. Quantification of the nuclear protein expression of NF-κB p65 and Nrf2 was analyzed by Image J. Lamin B was used as the internal control. d and e The expression of antioxidant associated proteins HO-1 and NQO-1 were measured by Western blot. Quantifications of proteins expression of HO-1 and NQO-1 were analyzed by Image J. GAPDH was used as loading and normalization control. f and g Quantitative RT-PCR determination of IL-6 and TNF-α mRNA and values normalized to GAPDH. h and i ELISA analysis for the concentrations of IL-6 and TNF-α in HK-2 cells supernatants. The values are expressed as the mean ± SD. *P < 0.05, **P < 0.01 versus the control group; ^#P < 0.05, ^##P < 0.01 versus the PA group; ^&P < 0.05, ^&&P < 0.01 versus the rhFGF21 group