2 μl/injection site) into the LPBN. They were then deeply anaesthetised with sodium thiopental (CRISTALIA, Itapira, SP, Brazil, 80 mg/kg of b.w.) and perfused transcardially with saline followed by 10% formalin. The brains were removed, fixed in 10% formalin, frozen, cut coronally into 60 μm sections and stained with Giemsa stain. Only animals with injections into the LPBN were considered for statistical analysis. The results are reported as means ± standard error of the mean (SEM). Statistical analysis was performed using two-way analysis of variance (ANOVA) with repeated measures followed by Student–Newman–Keuls post hoc tests to determine significant differences between
groups. IL-6 and TNF-α levels and alveolar bone loss were analysed by the Student Galunisertib price t-test. Differences were considered significant at P < 0.05. The software used to analyse the data was SigmaStat
this website for Windows, version 2.03 from SPSS Inc. Alveolar bone analysis revealed that rats with periodontal disease had more bone loss than rats in the control group (1.29 ± 0.04 vs. CN group: 0.50 ± 0.02 mm, P < 0.001, Fig. 1), showing that the induction of periodontal disease was effective. There were no statistically significant differences between the ingestion (ml/24 h) of water and 0.3 M NaCl for both groups (control and PD rats) in any of the evaluation periods (3 and 16 days) after ligature-induced periodontal disease (Fig. 2). ANOVA showed significant differences among treatments and times for water intake (F(18,126) = 17.4; P < 0.001, Fig. 3C and D) and 0.3 M NaCl intake (F(18,126) = 5.4; P < 0.001, Fig. 3A and B) when fluid-replete rats had simultaneous access to water and 0.3 M NaCl. Compared with saline injections into the LPBN, the cumulative ingestion of 0.3 M NaCl and water significantly increased after injections of muscimol (0.5 nmol/0.2 μl at each site, n = 8) into the LPBN after 120 min until the end of the test in control and PD rats ( Fig. 3A and
C). Post PRKACG hoc tests showed that ligature-induced PD attenuated the effects of muscimol on water intake ( Fig. 3C and D) without changing 0.3 M NaCl intake ( Fig. 3A and B). ANOVA showed significant differences among treatments and times for water intake (F(18,126) = 6.9; P < 0.001, Fig. 4C and D) and 0.3 M NaCl intake (F(18,126) = 4.7; P < 0.001, Fig. 4A and B) when FURO + CAP-treated rats (control and PD) that received muscimol or saline in the LPBN had simultaneous access to water and 0.3 M NaCl. In control rats, the cumulative ingestion of 0.3 M NaCl after injections of muscimol (0.5 nmol/0.2 μl at each site, n = 8) into the LPBN was significantly different from ingestion after saline injections into the LPBN from 90 to 180 min of the test, with P values ranging from P < 0.05 at 90 min to P < 0.005 from 120 to 180 min (Newman–Keuls post hoc test) ( Fig. 4A and B).