Study On The Effects Of Interleukin-23 And Interleukin-35 In Chronic Periodontitis And Clinical Case Report

Chronic periodontitis(CP)is a chronic inflammatory disease characterized by destruction of periodontal attachment apparatus including periodontal ligament,gingiva,alveolar bone and cementum with microbial infection.At present,the main indicators for the severity of periodontitis are:probing depth(PD),sulcus bleeding index(SBI)and X-ray,but these indicators can only represent a single lesion of the final results,and can not accurately reflect the occurrence,development and change of periodontitis.During the occurrence and development of periodontitis,in addition to bacterial microbial infection,the immune regulation of host was important,especially the immunomodulation effect of cytokines.Cytokines are biologically active proteins secreted by activated immune cells or other cells.They can activate cell signaling pathways,regulate cellular physiological functions,mediate inflammatory responses,and participate in immune responses and tissues repair and so on,by targeting cell receptors.Cytokines play an important role in promoting the inflammation of gum tissues,accelerating the destruction of periodontal connective tissue and the absorption of alveolar bone.So,to find effective cytokines which related to periodontitis development is very important for us to reveal the pathogenesis of periodontitis.This is also an important issues that needs to be solved urgently in early diagnosis and treatment of the periodontitis.Therefore,this project intends to combine previous and pre-work on the basis of periodontitis to study the role of IL-12 family members,IL-23 and IL-35,in periodontitis development and its related mechanisms,and to further understand the pathogenesis of periodontitis to lay the foundation for finding an effective periodontitis diagnosis and treatment technology.ObjectivesThe purpose of this study,we wondered whether the elevated IL-35 affected cytokine production from host innate immune cells in CP patients.We assay the expression of IL-23 and IL-35 in gingival tissue of patients with CP.In addition,the relationships among the aberrant expression of IL-35,IL-23 and sCD14 in gingival crevicular fluid(GCF)and serum from the CP patients after non-surgical periodontal therapy matched with the healthy volunteers were evaluated.The effect and related mechanism of IL-35,IL-23 in periodontitis development were also discussed.Materials and methodsThe research content of this study was divided into the following two parts:Part 1:Expression of IL-23 and IL-35 subunits in gingival tissues of patients with CP and its significance.1.The mRNA expression of IL-23(p19,p40)and IL-35(p35,EBIB)subunit in gingival tissue of patients with CP were detected by Real-time fluorescence quantitative PCR.2.The expression of IL-23(p19,p40)and IL-35(p35,EBI3)subunit in gingival tissue of patients with CP were detected by Western bloting.3.The relationship among IL-23,IL-35 and various clinical indicators were analyzed.Part 2:Detection of IL-23,IL-35 and sCD14 in gingival crevicular fluid and serum in patients with CP and its significance.1.The changes of clinical indicators before and after treatment were recorded.2.The levels of IL-23,IL-35 and sCD14 in gingival crevicular fluid and serum before and after treatment were measured.3.Relationship between IL-35,IL-23 and sCD14 levels and disease severity in CP patients were analyzed.4.Relationship of IL-35,IL-23 and sCD 14 levels between in gingival crevicular fluid and serum in CP patients were analyzed.5.The correlation among IL-35,IL-23 and sCD14 levels in CP patients were analyzed.Results1.The relative expression of pi 9,p35,p40 and EBI3 mRNA were higher on gingival tissue of CP patients than that of HV control.2.The expression of p19,p35,p40 and EBI3 protein were higher on gingival tissue of CP patients than that of HV control.3.The relative expression of p40 and EBI3 mRNA on gingival tissue of severe CP patients were higher than that of moderate CP.4.The relative expression of p35 mRNA was positive correlation with the relative expression of EBI3 mRNA on gingival tissue of CP patients.5.The relative expression of p19 mRNA was positive correlation with the relative expression of p40 mRNA on severe CP patients.6.The levels of IL-35,IL-23 and sCD14 were higher on GCF and blood of CP patients than HV control.7.The levels of IL-35,IL-23 and sCD14 were related to the degree of disease of CP patients.8.The levels of IL-35,IL-23 and sCD14 on GCF were positive correlation with those from blood before periodontal therapy on CP patients.9.Significant correlation between IL-35 and sCD14 were detected on GCF of CP patients.ConclusionsIn this study,we found that the levels of p19,p40,p35 and EBI3 were abnormalities in gingival tissues of CP patients.Especially,the levels of p19,p40,p35 and EBI3 mRNA and protein were higher in gingival tissue of CP patients than that of HV control.This results suggestted that p19,p40,p35 and EBI3 may mediated inflammatory response to involve in CP pathogenesis,and are associated with the severity of the disease.P40 is an important constituent subunit of IL-12 family of proinflammatory cytokines IL-12 and IL-23,and EBI3 is an important constituent subunit of IL-12 family of inflammatory cytokines IL-27 and IL-35.The relative expression of p40mRNA and EB3 mRNA in gingival tissues of patients with severe CP was significantly higher than that of patients with moderate CP.This result also suggestted that gingival inflammation and anti-inflammatory signals were active in severe CP patients.The relative expression of p19 mRNA,p40mRNA,p35mRNA and EBI3 mRNA in gingival tissue of 30 patients with CP,only p35mRNA was positively correlated with the relative expression of EBI3 mRNA.P35 and EBI3 are IL-35 subunits,suggesting that IL-35 mediated CP development,especially in patients with severe CP.IL-35 may be an important anti-inflammatory signal in gingival tissue of CP patients.P19 is a unique subunit of IL-23 and a high expression of p19 in CP patients,and the correlation shows that p19 was associated with another subunit p40 that makes up IL-23.However,in severe CP patients,p19 mRNA and p40 mRNA expression was positively correlated with the relative expression of IL-23,suggesting that IL-23 may also mediate the local inflammatory process in gingival tissue of CP patients.In addition,we found that the levels of IL-35,IL-23 and sCD14 were elevated in CP patients.IL-35,IL-23 and sCD14 from GCF positive correlation with those from blood before periodontal therapy on CP patients.Importantly,elevated IL-35-related sCD14 but not IL-23 is associated with the severity of CP.Together,our data suggested that IL-35 may facilitate sCD 14 to induce innate immune cells mediated inflammatory response to involve in CP pathogenesis.