| Objective: In this study we quantitatively analysis of the levels of tumor necrosis factor(TNF)α,interleukin(IL)6,IL-10 and high mobility group box1(HMGB-1)in serum from the patients,and the correlation of organoleptic rating(OR)and dental plaque index(PLI)of the patients with severe chronic periodontitis with halitosis;and the effects of porphyromonas gingivalis lipopolysaccharide(Pg-LPS)on the secretion of TNF-α,IL-1β,IL-6,IL-10,HMGB-1 and expression of corresponding cytokine mRNA in peripheral blood monocytes/macrophages from the patients with severe chronic periodontitis were discussed.The aim of this study is to understand the possible mechanism of chronic periodontal inflammation and stubborn halitosis,and to explore new therapeutic targets for chronic periodontitis and stubborn halitosis.Methods: The research subjects were included into the following five groups:(1)8 healthy subjects without periodontitis and halitosis(control group);(2)20 patients with severe chronic periodontitis and halitosis(group I);(3)12 cases with simple halitosis(group II);(4)12 cases with mild chronic periodontitis without halitosis(group III);(5)24 patients with severe chronic periodontitis were randomly divided into 2 subgroups: chronic periodontitis+Pg-LPS stimulation group(experimental group I)and chronic periodontitis group(experimental group II).Peripheral blood mononuclear cells were separated by density gradient centrifugation and followed by dish adherence for monocytes/macrophages.The OR was determined by organoleptic method and PLI was measured by plaque disclosing agent.The levels of TNF-α,IL-1β,IL-6,IL-10 and HMGB-1 in peripheral blood and culture supernatant of monocytes/macrophages were examined with double antibody sandwich ELISA.The relative quantity for mRNA of TNF-α,IL-1β,IL-6,IL-10 and HMGB-1 in monocytes/macrophages were detected by RT-PCR.The statistical analyses were completed with SPSS 17.0.Results:(1)The levels of TNF-α,IL-6 and HMGB-1 in peripheral blood from individuals of group I and III were higher than those of control group(P<0.05).The levels of TNF-α,IL-6,IL-10 and HMGB-1 in peripheral blood from individuals of group I were higher than those of group II(P<0.05).The levels of HMGB-1 in group I was higher than those of group II and group III,and group III was higher than those of group II(P<0.05).(2)The OR level in individuals of group II was higher than those in group III(P<0.05).In groups I,II and III,OR was positively correlated with PLI(P<0.05).In group I,OR and PLI were positively correlated with the levels of TNF-α,IL-6 and HMGB-1(P<0.05).In group II,PLI was positively correlated with HMGB-1(P<0.05).In group III,the levels of OR and PLI were positively correlated with HMGB-1(P<0.05).(3)The release of cytokines(TNF-α,IL-1β,IL-6 and HMGB-1)from cultured peripheral blood monocytes/macrophages and the expression of corresponding mRNAs were significantly elevated after Pg-LPS stimulation in patients with severe chronic periodontitis(P<0.05).Conclusions:(1)The levels of TNF-α,IL-6 and HMGB-1 in peripheral blood from the patients with severe chronic periodontitis and halitosis were significantly higher than those from normal subjects,and the levels were positively correlated with readings of OR and PLI.(2)The over-expression and excessive release of inflammatory cytokines including TNF-α,IL-6,HMGB-1 and the relatively low expression of anti-inflammatory cytokine IL-10 may be the important factors in the pathogenesis of severe chronic periodontitis and stubborn bad breath.(3)Monocytes/macrophages in peripheral blood from the patients with chronic periodontitis may be constantly stimulated by oral antigens that resulted in a hypersensitive state,so the extremely low concentration of Pg-LPS stimulation can make the monocytes/macrophages overexpression and release of inflammatory cytokines which might be used as risk factors for aggravating inflammation response and inducing inflammation-related diseases. |
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