Overexpression of PKCζ and PKCι/λ impaired the insulin-regulated gene expression in ZL primary rat hepatocytes.

<p>Primary hepatocytes of ZL rats fed chow for 8 weeks were isolated and seeded onto dishes as described in the Materials and Methods. Purified Ad-β-gal, Ad-Prkcz and Ad-Prkci were added in medium A during the overnight pretreatment period to allow the overexpression of β-GAL, PKCζ and PKCι/λ, respectively. The primary hepatocytes were then incubated in fresh medium A with increasing concentrations of insulin (0nM to 100nM) for 6 hours. Total RNA was extracted, and subjected to real-time PCR analysis. The transcript level of each indicated gene in primary hepatocytes transfected with Ad-β-gal in the absence of insulin treatment was arbitrarily set to 1 for that gene. The mRNA levels of (A) <i>Gck</i>, (B) <i>Srebp-1c</i>, (C) <i>Pck1</i>, (D) <i>Igfbp-1</i>, (E) <i>G6pc</i> and (F) <i>Pklr</i> were expressed as fold inductions (mean ± SEM; n = 4; all p<0.05; af, g>h, i>j, k>l, m>n, o>p, q>r for comparing the insulin treatment groups of hepatocytes overexpressing Ad-β-gal, Ad-Prkcz or Ad-Prkci using one-way ANOVA; * for comparing fold inductions among Ad-β-gal, Ad-Prkcz and Ad-Prkci groups at the indicated insulin concentrations using one-way ANOVA).