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Protective effects of mercaptoethylguanidine,a selective inhibitor of inducible nitric oxide synthase,in ligature-induced periodontitis in the rat
Authors:Zsolt Lohinai  Péter Benedek  Erzsébet Fehér  Adrienn Gy?rfi  László Rosivall  árpád Fazekas  Andrew L Salzman  Csaba Szabó
Affiliation:*Experimental Research Department and 2nd Institute of Physiology, Semmelweis Univ. Med. School, Budapest, Hungary;?Department of Pathophysiology, Semmelweis Univ. Med. School, Budapest, Hungary;?Department of Anatomy, Semmelweis Univ. Med. School, Budapest, Hungary;#Clinic of Conservative Dentistry, Semmelweis Univ. Med. School, Budapest, Hungary;§Division of Critical Care, Children''s Hospital Medical Center, Cincinnati, U.S.A.
Abstract:
  1. Excessive production of nitric oxide (NO), and the generation of peroxynitrite have been implicated in various proinflammatory conditions. In the present study, using mercaptoethylguanidine (MEG), a selective inhibitor of iNOS and a peroxynitrite scavenger, we investigated the role of iNOS and peroxynitrite in a rat model of periodontitis.
  2. Periodontitis was produced in rat by a ligature of 2/0 braided silk placed around the cervix of the lower left 1st molar. Animals were then divided into two groups: one group of rats was treated with MEG (30 mg kg−1, i.p., 4 times per day for 8 days), animals in the other group received vehicle. At day 8, the gingivomucosal tissue encircling the mandibular 1st molars was removed on both sides from ligated and sham operated animals for inducible nitric oxide synthase (iNOS) activity assay and for immunocytochemistry with anti-iNOS serum. Plasma extravasation was measured with the Evans blue technique. Alveolar bone loss was measured with a videomicroscopy.
  3. Ligation caused a significant, more than 3 fold increase in the gingival iNOS activity, whereas it did not affect iNOS activity on the contralateral side, when compared to sham-operated animals. Immunohistochemical analysis revealed iNOS-positive macrophages, lymphocytes and PMNs in the connective tissue and immunoreactive basal layers of epithelium on side of the ligature, and only a few iNOS-reactive connective tissue cells on the contralateral side. Ligation significantly increased Evans blue extravasation in gingivomucosal tissue and alveolar bone destruction compared to the contralateral side. MEG treatment significantly reduced the plasma extravasation and bone destruction.
  4. The present results demonstrated that ligature-induced periodontitis increases local NO production and that MEG treatment protects against the associated extravasation and bone destruction. Based on the present data, we propose that enhanced formation of NO and peroxynitrite plays a significant role in the pathogenesis of periodontitis.
Keywords:Ligature   inducible nitric oxide synthase   nitric oxide   periodontitis   gingivitis   lipopolysaccharide   cytokine   tissue destruction
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