Fig. 5.

Effect of KBH-1 on leptin-resistant cortical neuron cells. Cells were cultured in neurobasal medium supplemented with 2% B27, 0.5 mM L-glutamine, and 25 μM glutamate. After 24 h, the culture medium was replaced with neurobasal medium without glutamate. For the resistance experiments, primary neurons were pretreated with 0.5 mM FFA (oleic acid:palmitic acid = 2:1) overnight prior to leptin treatment. For treatment studies, neurons were treated with KBH-1 for 1 h, then treated with 10 nM leptin. (a) Cell viability was determined by the CCK assay. (b) The activation of AMPK, RAS, cRaf, ERK, STAT3 and JAK2 were measured using SDS-PAGE and immunoblotting. β-actin was used for normalization (c, n = 3). The data are expressed as the mean ± SEM. Significant differences from each control (no KBH-1 treatment) are indicated by *p < 0.05 or **p < 0.001