The primer sequences for ApoB100 (forward
primer 5-AGTAGTGGTGCGTCTTGGATCCA-3′ and reverse primer 5-ACTCTGCAGCAAGCTGTTGAATGT-3′) were derived from the Rattus norvegicus genome (National Center for Biotechnology Information GenBank, accession number NM_019287) and were constructed using the Primer-BLAST Program (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). The forward and reverse primer sequences for LDL-R and HMG CoA-R were obtained from published nucleotide Quizartinib purchase sequences [35], as were those for glyceraldehyde-3-phosphate dehydrogenase [36]. All primers were synthesized by Invitrogen Life Technologies (São Paulo, Brazil). The reactions were performed using an ABI Prism 7000 Sequence Detector (Applied Biosystems) under the following conditions: 50°C for 2 minutes, 95°C C59 wnt chemical structure for 10 minutes, and 40 cycles of 95°C for 15 seconds, and 60°C for 1 minute. The specificity of the products obtained was confirmed by analyzing the dissociation curves of the amplified product.
As an internal control, the expression of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene was used. The data obtained were analyzed using the comparative cycle threshold method. All analyses were performed in triplicate. The normality of the data was tested using the Kolmogorov-Smirnov test. Data (Table 2, Table 3 and Table 4) consistent with a normal distribution were subjected to 2-way analysis of variance in which the classification factors were diet (C + CA × H + HA), açaí (CA + HA × C + H), and the interaction between diet and açaí (C × CA × H × HA). The Bonferroni t test was used for multiple comparisons among the means. Data that did not fit the normal distribution were analyzed using a Kruskal-Wallis nonparametric test and Dunn posttest. The differences were considered statistically significant when P < .05. For the remaining analyses ( Fig.), Student unpaired t test was used. The results are expressed as means and SDs or as medians and interquartile ranges. The minimum sample size needed to detect
a statistically Sitaxentan significant difference (P < .05) was calculated based on the power of 0.9 (G*Power 3.13, statistical power analyses program; http://www.psycho.uni-deusselforf.de/aap). Statistical analyses were performed using GraphPad Prism version 4.00 for Windows (GraphPad, San Diego, CA, USA). We first examined how the addition of açaí pulp in the diet affected body weight gain, liver weight, fecal excretion, and food intake. The data in Table 2 indicate that hypercholesterolemic rats exhibited an increase in weight gain and liver weight. The addition of 2% açaí pulp to the diets did not affect these parameters. The rats of the H group ingested less food and excreted a lower amount of feces compared with the controls.