Thus, a better understanding of negative regulatory mechanisms of JAK/STAT pathway during inflammatory response may lead to important information on periodontal disease pathogenesis and also provide a therapeutic selleck chemical perspective based on the modulation of pro-inflammatory gene expression. This study evaluated the kinetics of SOCS1 and SOCS3 expression in ligature-induced model of periodontal disease in rats. We also evaluated the

mRNA expression of TNF-α, IL-6 and IL-10 that are direct targets of SOCS proteins and the mRNA expression of RANKL, OPG and that were shown to be relevant for pathogenesis of periodontal disease and may be indirect targets of SOCS proteins. Male adult Wistar (Norvegicus albinus) rats (N = 36) were obtained from the S6 Kinase inhibitor Multidisciplinary Center for Biological Investigation (CEMIB-UNICAMP). The animals, weighing approximately 250 g each, were maintained with food and water ad libitum. The experimental protocol was approved by the Ethical Committee on Animal Experimentation (protocol number 23/2007) of the School of Dentistry at Araraquara –

UNESP and performed in accordance with the guidelines from the Brazilian College for Animal Experimentation (COBEA). General anesthesia was induced with intramuscular injections of ketamine and xylazine chloridrate at 0.08 mL/100 g body weight and 0.04 mL/100 g body weight, respectively. The animals were divided into two experimental groups: A – sham-operated group (n = 9) – animals were anaesthetised but no ligatures were placed on the lower molars B – experimental group (n = 27) – a cotton thread ligature was placed around the cervical area of the lower first molars bilaterally to induce experimental periodontal disease. After 7, 15, and 30 days of the ligature placement (baseline), 3 animals from the control group and 9 animals from the experimental group were sacrificed per period by anesthetic overdose. The mandibular jaws were hemisected, and half of the block samples including molars with their surrounding tissues were submitted to routine

histological processing to be used in the stereometric Thalidomide evaluation. The other half of the blocks had the gingival tissue around the first molars carefully dissected for extraction of total RNA and protein for RT-qPCR and western blot analysis. After dissection of the gingival tissues, the samples were immersed in 3% hydrogen peroxide for 24 h to remove remaining soft tissues. Subsequently, these samples were stored in 70% ethanol and used for the macroscopic assessment of bone resorption. The area of bone resorption in the lingual surface of the first molars was measured macroscopically. Briefly, the pieces were removed from alcohol, dried, immersed for 5 min in a solution containing 0.7 g/L of methylene blue and washed with tap water to remove the excess dye.