A Matricaria recutita previne a perda óssea alveolar osteoclástica induzida por ligadura em ratos via inibição das citocinas TNF-a E IL-1b

Abstract

Periodontitis is an immunoinflammatory disease in that the involvement of chemical mediators culminates in destruction of alveolar bone. Long recognized regarding its pathogenesis, however, frequently some patients do not respond insatisfactorily to conventional treatments, which makes pharmacological alternatives are sought. In this context, Matricaria recutita (MTR), known as chamomile, stands out in the literature for its anti-inflammatory and a variety of constituents, especially apigenin (APG) flavonoid. Thus, the present study evaluated the involvement of cytokines in the anti-inflammatory and antiresorptive activities of MTR in alveolar bone resorption (ABR) induced by ligature in rats. For this, we used the dry extract of MTR (apigenin content 128.5±0.99 mg/g). The ABR was induced in 90 wistar rats (199.3 ± 3.2 g) by ligation (nylon 3.0) of 2° upper left molar, and contralateral was used as control. The rats received v.o. Tween 80 (TW) or MTR (10, 30 and 90 mg/kg) daily until 11 d, when they were killed. The hemiarcadas were processed for macroscopic (mm2) or histometric, histological and immunohistochemical analyzes for the ligand of the receptor activator of nuclear factor kappa B (RANKL), osteoprotegerin (OPG) and tartrate-resistant acid phosphatase (TRAP). Blood samples were collected for measurement of bone alkaline phosphatase (BALP), while the gingival tissue was used for measuring of mieloperoxidase activity (MPO; mg/g) and of tumor necrosis factor-alpha (TNF-a) and interleukin-1β (IL-1β) levels (pg/mg) by ELISA. Systemically, serum bone alkaline phosphatase (BALP), AST/ALT, urea and creatinine, and white blood count were made, and we evaluated of macroscopic aspects of liver, kidneys and spleen, in addition to variation in body mass. Was set at p <0.05 (#) for Normal, (*) for TW and () for MTR 10 mg/kg; Ethical aspects: the Ethics Committee for Animal Use-UFC 70/13. It was found that ligation for 11 days caused intense ABR with furcation lesion pronounced, resorption of alveolar bone and cementum in the region between the first and second molars, reduction of serum BALP, intense leukocyte infiltrate in the periodontium these animals, increasing significant MPO, TNF-, IL-1β in challenged area underlying gingival tissue, and increased to RANKL and TRAP immunostaining, and reduced to OPG. Systemically, there was leukocytosis with a predominance of mononuclear cells. No major changes in organs and weight of animals were observed. MTR prevented, significantly, the ligature-induced ABR [TW=5.5±0.2; MTR (10)=4.4±0.1*; (30)=2.9±0.1*; (90)=2.8±0*], corroborating the reduction of furcation lesions [Normal=10.4±0.8; TW=137.4±23.3#; MTR (90)=81.0±9.6*#] and the preservation of the alveolar bone and cementum [(Normal=0(0-0); TW=3(1-3)#, MTR (90)=1(1-3)#*] compared to the TW group while no bone anabolic activity was showed because MTR dit not prevent the reduction of serum BALP induced by ligature [Normal=99.4±3.4; TW=61.3±2.6#; MTR (10)=70.6±3.6#; (30)=74.5±3.7#; (90)=78.5±2.8#); p>0.05]. However, MTR significantly prevented the leukocyte infiltration and the increase of MPO activity [Normal=3.6±0.5; TW=9.4±0.9#; MTR (10)=10.2±3.3; (30)=4.5±0.8*; (90)=4.2±0.7*], of TNF-a [Normal=0.2±0; TW=1.2±0.2#; MTR (10)=0.4±0.2*; (30)=0.2±0.1*; (90)=0.1±0*] and of IL-1β [Normal=1.5±0.3; TW=8.0±1.4#; MTR (10)=8.9±1.9#; (30)=1.8±1.0*; (90)=1.5±0.9*] levels caused by ligature, and reduced immunostaining for RANKL and TRAP, and increased for OPG, comparing to TW group. Additionally, MTR prevented the leukocytosis caused by ligation and did not alter liver, kidney, spleen conditions or the variation of body mass. In short, the MTR prevented the ABR by reducing TNF-a and IL-1β, thus preventing the osteoclast activation due RANK-RANKL-OPG axis, without interfering with bone anabolism.