| BACKGROUND:Peri-implantitis,which is characterized by dense inflammatory infiltrates and increased osteoclast activity,can lead to alveolar bone destruction and implantation failure.However,the pathogenesis of peri-implantitis has not been completely clarified.With the development of high-throughput sequencing technology,the application of transcriptome research methods provides new ideas for studying the mechanism of diseases.This suggests that we could further reveal the pathogenesis of peri-implantitis from the perspective of transcriptome studies.OBJECTIVE:By studying the mRNA and miRNA genome expression profiles of the surrounding soft tissues of peri-implantitis,We constructed the miRNA-mRNA co-expression network.The potential promoter factors associated with immunity to peri-implantitis were explored,and the key genes were screened out,providing the theoretical basis for subsequent experimental verification.METHOD:1.Soft tissue samples of peri-implantitis patients and normal patients admitted to the department of stomatology of Guangxi Medical University were collected.The mRNA and miRNA libraries were established for the next step of PCR fragment amplification for the total RNA that passed the quality test.Then,Illumina Hiseq platform was used to carry out 2×150 double-terminal sequencing for the sample libraries that passed the quality control.2.The mRNA and miRNA differential gene expression profiles obtained by sequencing were used for screening of differentially expressed genes(DEGs and miDEGs),gene ontology,KEGG pathway enrichment analysis,protein-protein network interaction analysis,screening of key genes and network module analysis,and further correlation analysis of miRNA-mRNA differential genes.RESULTS:1.Results of mRNA differential expression gene analysis(1)A total of 1949 DEGs(861 upregulated and 1088 downregulated)were identified in the peri-implantitis samples compared with normal ones.(2)The GO terms of upregulated DEGs were mainly associated with reaction to molecules of bacterial origin,cell adhesion and immune response,extracellular space,endoplasmic reticulum cavity,cytokine activation and peroxidase(POD)activation.The downregulated DEGs were mainly enriched in the skin development,epidermal development,tissue development,intrinsic components of the plasma membrane,extracellular matrix containing collagen and the activation of ion transmembrane transporters.(3)Four pathways that were significantly enriched by upregulated DEGs were cytokine and receptor interactions,staphylococcus aureus infection,osteoclast differentiation and chemokines.The downregulated DEGs were mainly enriched in circadian rhythm and transforming growth factor-signaling pathway.(4)the PPI network was constructed with 1,398 nodes and 1,537 protein pairs.GNGT2,GNG7,C3,PPBP,IL6 and HSP90B was the hub genes in the PPI network.GNGT2,GNG7,C3 and PPBP,hub genes with higher node degrees in module 1,were significantly enriched in immune response of biological processes.In addition,IL6 and HSP90B were enriched in bone remodeling regulation and bone resorption of biological process in module 6.2.Analysis results of miRNA differentially expressed genes(1)A total of 204 miDEGs(114 upregulated and 90 downregulated)were identified in the peri-implantitis samples compared with normal ones.(2)The GO terms of miRNA target genes showed that the upregulated target genes were mainly associated with the cascade regulation of ERK1/ERK2,intercellular adhesion and other biological processes related to immunity.Downregulated target genes are mainly enriched in biological processes such as regulation and cell differentiation.(3)KEGG pathway showed that the unregulated target genes were significantly enriched in the pathways related to immunity and inflammation,such as lysosome,natural killer cell-mediated cytotoxicity and osteoclast differentiation.Down-regulated target genes were considerably enriched in cancer-related signaling pathways.(4)The results of enrichment analysis revealed that miR-3679-5p and miR-508-5p may play important roles in peri-implantitis and are worth further investigation.3.Correlation analysis of miRNA-mRNA(1)The mRNA-miRNA interaction pairs consisted of 142 interaction pairs between 109 DEGs and 50 miRNAs.(2)miR-4446-3p is the center of the whole network and may inhibit peri implant osteogenesis by reversely regulating its target gene FGFR2.CONCLUSION:1.GNGT2,GNG7,C3,PPBP,IL6 and HSP90B were predicted to be biomarkers of peri-implantitis.2.miR-4446-3p,miR-3679-5p and miR-508-5p may be involved in the pathogenesis of peri-implantitis by reversely regulating their target genes. |
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