24 Digital images were analysed using Sigma-Scan 2.0 software. The distance between the cemento–enamel junctions up to the height of alveolar bone of the first mandibular molar on the mesial side of the rat was recorded. Samples were homogenised in Trizol reagent (Invitrogen) for 1 min using a tissue homogenizer (Polytron-Agrgregate, Kinematica, Littau/Luzern, Switzerland) at maximum speed. Total RNA was isolated according to the manufacturer’s guidelines and quantified by a spectrophotometer. Cytoskeletal Signaling inhibitor The integrity of RNA was verified by agarose gel electrophoresis. Complementary DNA was prepared using 2 μg of total RNA and a reverse transcriptase. The primers used
in the experiments were the standard TaqMan (Applied Biosystems, Foster City, CA, USA) brand. The gene analysed were TNF-α
(primers: sense 5′ GGC ATG GAT CTC AAA GAC AAC C-3′ and antisense 5′-CAA ATC GGC TGA CGG TGT G-3′). Glyceraldehyde-3-phosphate dehydrogenase (GenBank NM_017008) was used as a housekeeping gene. Real-time polymerase chain reaction was carried out in the StepOne polymerase chain reaction cycler (Applied Biosystems, Foster City, CA, USA). The polymerase chain reaction conditions were 95 °C for 10 min, followed by 40 cycles at 95° for 10 s and 60 °C for 45 s. Real-time data were analysed using the Sequence Detector System 1.7 (Applied Biosystems, Foster City, CA, USA). Results are expressed as fold inductions compared with controls. Results www.selleckchem.com/products/Cyclopamine.html are presented as means ± SEM for the number of rats (n) indicated. The data were analysed by the unpaired Student’s t-test for two mean comparisons and one-way ANOVA (with Bonferroni post hoc test) for bone reabsorption and TNF-α expression. The level of significance was set at P < 0.05. Table 1 shows that the body weight and naso-anal length were 29% and 15% respectively, Dipeptidyl peptidase lower in MSG groups when compared with CTL (P < 0.05), however, the Lee Index was 8% higher in the MSG rats (P < 0.003). The retroperitoneal and perigonadal fat pads weight doubled in
MSG rats when compared with CTL rats (P < 0.0001, Fig. 1A and B). The neonatal MSG treatment did not influence the plasma concentration of glucose, NEFA and total CHOL (P > 0.05). However, in the MSG group plasma and TG concentrations were 3.0 and 4.0 times higher (P < 0.0001 and P < 0.0002), respectively than, CTL group ( Table 2). According to Fig. 2, alveolar bone resorption was 44% lower in obese-MSG group compared with CTL group (P < 0.01). In the presence of ligature, there was a significant increase in alveolar bone resorption in both groups CTL L and MSG L compared with CTL and MSG group respectively (P < 0.001). However, alveolar bone resorption in the MSG L animals was similar to that occurring in the CTL group (P > 0.05) ( Fig. 3A–D). The TNF-α gene expression in periodontal tissue was similar in MSG and CTL animals in the absence of ligature (P > 0.