Comparative Study On The Pathogenesis Of Peri-implantitis And Periodontitis Based On The Preliminary Analysis Of Whole Transcriptome Sequencing

BACKGROUND:Although periimplantitis and periodontitis share similar features,particularly pathogeny,clinical features and treatment,however,more and more evidences show that there are some differences they are two different diseases and should be analyzed separately.With the rapid development of high-throughput sequencing technology and bioinformatics,non codingRNA(ncRNA)has abnormal expression in a variety of inflammatory diseases,including periodontitis,and has a good prognostic value,but there are few related studies in peri-implantitis.It is suggested that we can make a further comparative study on the pathogenesis of peri-implantitis and periodontitis from the perspective of transcriptome.OBJECTIVE:Based on the whole transcriptome sequencing technology,this study constructed an endogenous competitive network(ceRNA)mechanism analysis by studying the expression profile of the whole transcriptome in peri-implantitis and periodontitis lesions,and revealed the different effects of peri-implantitis and periodontitis from two aspects of molecular and biological processes,and preliminarily explored its metabolism,signal transduction,cell fate and epigenetic regulation The different pathological processes and possible mechanisms of periodontitis provide new ideas for further study of the similarities and differences between peri-implantitis and periodontitis.METHODS:1.The gingival samples of peri-implantitis(PI),periodontitis patients(P)and healthy individuals(HI)were collected.TotalRNAs and miRNA libraries were established for PCR amplification,and then the qualified sample libraries were sequenced 2 × 150 by using Illumina hiseq platform.2.The gene expression profiles of mRNA,miRNA and lncRNA were obtained by sequencing.R software and Metascape were used to screen differentially expressed genes,to analyze GO function enrichment,to analyze KEGG pathway enrichment and to analyze protein interaction network.3.Combined with the analysis of differentially expressed genes,database prediction and correlation,the mRNA-miRNA-lncRNA relationship pair was constructed,and the Cytoscape was used to construct the ceRNA interaction network.Finally,gene set enrichment analysis(GSEA)was used to predict the function of key nodes in the CeRNA network.RESULTA:1.In this study,a total of 15 gingival samples were collected,including 5cases of peri-implantitis(PI),5 cases of periodontitis(P)and 5 cases of healthy individuals(HI).2.Differential expression analysis of mRNAs、miRNAs、lncRNAs FromRNA-seq.(1)In this study,139 significantly up-regulated mRNAs and 2753down-regulated mRNAs,165 significantly up-regulated miRNAs and 37down-regulated miRNAs,and 631 significantly up-regulated lncRNAs and 2901down-regulated lncRNAs in the PI-vs-P group.(2)Muscle filament sliding,oxygen transport and regulation of neurological system process,immune system response were enrichment of the DEGs in PI-vs-P.Further analysis of the relationship between significant membership term and gene expression showed that PI-vs-HI group and PI-vs-P group were significantly enriched in the pathway and process of metal ion response and innate immune response.P-vs-HI,PI-vs-HI and PI-vs-P groups were significantly enriched in defense response and reactive oxygen species.In these four pathways and processes(response to metal ions,innate immune response,defense response and reactive oxygen species response),the expression level and distribution of different genes involved in PI and P groups were significantly different(P < 0.05),and each pathway and process involved in The level of gene expression was significantly higher than that of P group.(3)Through the construction of protein interaction network,181 nodes and300 protein pairs were obtained.10 modules were identified by MCODE analysis.Further analysis showed that PI-vs-P group was significantly enriched in modules 1,2,5,6 and 8,module 1 was mainly involved in the biological process of cytochrome P450,the first stage functionalization of compounds and biological oxidation;module 2 and module 5 were mainly involved in myofilament sliding,actin myosin filament sliding,actin mediated cell contraction and striated muscle contraction;module 6 was mainly involved in MHC class II Preparation and expression of antigens,preparation of MHC class II antigens,presentation of peptide antigens,antigen treatment and presentation of peptide or polysaccharide antigens via MHC class II;module 8 is mainly involved in the biological process of endocytosis and membrane transport mediated by grid proteins.3.Analysis of the ceRNA network(1)Construction of ceRNA network: in this study,593 pairs ofRNAs were obtained,including 16 miRNAs,91 lncRNAs and 377 mRNA nodes.(2)Topology analysis of ceRNA network: 6 mRNAs(FAM126B,SORL1,PRLR,CPEB2,RAP2 C,YOD1),5 lncRNAs(lnc-CORO2B-1,lnc-MBL2-7,lnc-TRIM45-1,lnc-CHST10-2,lnc-TNP1-6),7 miRNAs(hsa-mi R-544 a,hsa-mi R-9-5p,hsa-mi R-650,hsa-mi R-338-5p,hsa-mi R-3179,hsa-mi R-149-5p,hsa-mi R-196a-5p).The aboveRNA molecules and their interaction are the potential regulatory mechanisms of ceRNA in peri-implantitis and periodontitis.(3)The GSEA results of six key mRNAs in the ceRNA network showed that fam12 b and CPEB were mainly involved in T cell receptor signaling,Wnt signaling,toll like receptor signaling and nod signaling,JAK / MAPK signaling,RIG-I signaling and apoptosis related pathways.Rap 2C and yod1 not only participate in the regulatory pathways similar to fam12 b and CPEB,but also are significantly related to the Notch signaling pathway.SORL1 and PRLR were significantly correlated with Wnt signaling pathway and oxidative phosphorylation process.CONCLUSION1.There were significant differences expression profiles of mRNA,miRNA and lncRNA between peri-implantitis patients and periodontitis patients.2.Six mRNAs(FAM126B,SORL1,PRLR,CPEB2,RAP2 C,YOD1)and five lncrnas(lnc-CORO2B-1,lnc-MBL2-7,lnc-TRIM45-1,lnc-CHST10-2,lnc-TNP1-6),7 miRNAs(hsa-mi R-544 a,hsa-mi R-9-5p,hsa-mi R-650,hsa-mi R-338-5p,hsa-mi R-3179,hsa-mi R-149-5p,hsa-mi R-196a-5p)of ceRNA,and their lncRNA-miRNA-mRNA interaction can be used as the potential regulatory mechanism of ceRNA in peri-implantitis and periodontitis.The interaction of lncRNA-miRNA-mRNA involves the regulation of Wnt,Notch,Hippo and other proliferation signaling pathways,ROS related processes,Toll like receptor signaling,NOD signaling pathways and host stress processes.Unlike periodontitis,these signaling pathways and biological processes may be the main factors affecting the pathogenesis of peri-implantitis.