0022, respectively). The highest levels of serum TG, T-CHOL, and HDL-C were found in the TFA+MSG Ivacaftor group, with increases of 39.3, 10.5, and 42.8%, respectively (P �� 0.05). Fasting blood glucose, insulin, leptin, adiponectin, and RBP4 were measured (Table 2). There was no significant difference in fasting glucose and adiponectin levels. Fasting insulin was significantly elevated in the MSG group (P < 0.05, n = 6). Serum leptin levels were markedly elevated in the TFA and TFA+MSG diet groups (P = 0.0138, n = 6). Conversely, serum levels of RBP4 were elevated in all three diet groups relative to control (P �� 0.001). Hepatic histology, lipid staining, and TG content Increased hepatic lipid deposition as depicted with Oil-Red-O staining was apparent in 32-week MSG- and TFA-treated animals compared with control (Fig.

1A). The livers of TFA-treated animals contained larger vesicular structures consistent with macrovesicular steatosis (Fig. 1A, B). Conversely, microvesicular steatosis was apparent in the livers of MSG-treated animals, as indicated by Oils-Red-O (Fig. 1A) and HE staining (Fig. 1B) of mouse liver tissue. Interestingly, in the TFA+MSG animals, microvesicular steatosis was predominant, and there was an attenuation of macrovesicular structures in the livers from this diet group (Fig. 1B). Higher magnification revealed an increase in hepatic cell volume and nuclear size in MSG-treated livers (Fig. 1C). While the weight of livers from control, MSG, and TFA diet group mice did not differ significantly, livers from TFA+MSG-treated mice weighed an average 1.

54-fold higher than control (Fig. 1D; P �� 0.05, n = 10). Intrahepatic TG levels were quantified (Fig. 1E). The highest level of hepatic TG occurred in the TFA diet group with a 1.43-fold increase in hepatic lipid content compared with control (Fig. 1E; P �� 0.05, n = 6). Fig. 1. Diet-induced changes in liver histology and TG content. A: Increased lipid deposition, as indicated by Oil-Red-O lipid staining in liver sections from mice on the MSG, TFA, and TFA+MSG diet groups at 32 weeks, compared with control. Red staining indicates … Hepatic gene expression To detect early changes in gene expression that might account for the hepatic steatosis and dyslipidemia seen in 32-week-old mice, total RNA samples were isolated from liver tissue obtained from four individual 16-week-old mice from each of the diet groups and pooled (n = 2) for applying onto two individual Affymetrix Mouse Genome 1.

0 ST Array Expression Set GeneChips containing 28,853 genes. Statistical analysis of these two sets of paired samples using a t-test (P �� 0.05, n = 4) identified a group of 84 hepatic genes whose expression was significantly changed within the four diet groups. Figure 2 shows the heat map and clustering dendogram of the differentially Brefeldin_A expressed genes. Table 3 shows 84 genes falling into 12 different functional ontologies, which were differentially expressed within the four groups, using a 1.