种植体周围炎炎性组织差异表达基因的筛选及验证  

Screening and verification of differentially expressed genes in inflammatory tissues with peri-implantitis

作　　者：韦雨杏 董皓[2] 韦惠平[2] 谢诚 陀杨娟 覃浩 曹勇[1,2] Wei Yuxing;Dong Hao;Wei Huiping;Xie Cheng;Tuo Yangjuan;Qin Hao;Cao Yong(College of Stomatology/Hospital of Stomatology,Guangxi Medical University,Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Guangxi Clinical Research Center for Craniofacial Deformity,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of Oral and Maxillofacial Surgery,Guangxi Academy of Medical Sciences/People’s Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学口腔医学院/附属口腔医院,广西口腔颌面修复与重建研究重点实验室,广西颅颌面畸形临床医学研究中心,广西壮族自治区南宁市530021 [2]广西医学科学院/广西壮族自治区人民医院口腔颌面外科,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2023年第30期4844-4849,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金(2018GXNSFAA050088),项目负责人:曹勇;广西医疗卫生适宜技术开发与推广应用项目(S2021048),项目负责人:曹勇。

摘　　要：背景:种植体周围炎严重影响着口腔种植修复的成功,目前仍然没有有效的治愈方法,其发病机制也尚未阐明。目的:寻找种植体周围炎发生相关的核心差异表达基因,并进行实验验证。方法:利用生物信息技术从GEO数据库中筛选种植体周围炎牙龈和健康牙龈的差异表达基因,并对差异基因进行富集分析。利用String数据库构建蛋白互作网络,并使用cytoscape软件筛选关键基因,采用RT-PCR和免疫组化检测对关键基因C3AR1表达进行验证。采用RT-PCR法检测RAW264.7细胞破骨向分化后C3AR1 mRNA的表达水平。结果与结论:①将GSE33774数据集中种植体周围炎牙龈和健康牙龈的基因数据行差异分析,得到有意义的上调基因33个,下调基因17个,共50个差异表达基因;GO和KEGG结果显示差异表达基因主要富集在免疫受体活性、C-C趋化因子受体活性功能,以及破骨细胞分化相关信号通路上;②共筛选出C3AR1、CD14、TLR4、CCR1、CYBB和FCGR2A 6个关键基因,RT-PCR、免疫组化实验结果证实,C3AR1在种植体周围炎组织中表达上调,且C3AR1在巨噬细胞破骨向分化后表达上调;③结果显示,C3AR1在种植体周围炎中高表达,可能是种植体周围炎骨质破坏的关键基因。BACKGROUND:Peri-implantitis seriously affects the success of dental implant restorations.There is still no effective cure and its pathogenesis has not yet been elucidated.OBJECTIVE:To find key differentially expressed genes related to the occurrence of peri-implantitis and experimentally verify them.METHODS:Differentially expressed genes between the peri-implantitis and the healthy gums were screened from the GEO database using bioinformatics technology and were analyzed by enrichment analysis.A protein interaction network was constructed using the String database.Key genes were screened by Cytoscape software.Expression of key gene C3AR1 was validated using RT-PCR and immunohistochemistry.The expression level of C3AR1 mRNA in RAW264.7 cells after osteoclast differentiation was detected by RT-PCR.RESULTS AND CONCLUSION:(1)The gene data of peri-implantitis and the healthy gums in the GSE33774 data set were analyzed for differences.A total of 50 differentially expressed genes were obtained,including 33 upregulated genes and 17 downregulated genes.GO functional enrichment and KEGG pathway enrichment analysis showed that differentially expressed genes mainly played a role in immune receptor activity,C-C chemokine receptor activity,and the signaling pathway of osteoclast differentiation.(2)Six key target genes were identified based on the protein interaction network,including C3AR1,CD14,TLR4,CCR1,CYBB,and FCGR2A.The results of RT-PCR and immunohistochemistry experiments showed that the expression of C3AR1 at mRNA and protein levels was significantly increased in peri-implantitis tissue.Moreover,C3AR1 was upregulated after osteoclastic differentiation of macrophages.(3)It is concluded that C3AR1 showed high expression in peri-implantitis,which may be a key gene for bone destruction in peri-implantitis.

关 键 词：种植体周围炎 C3AR1 生物信息学 差异表达基因 关键基因 单核巨噬细胞 破骨分化 骨质破坏 

分 类 号：R459.9[医药卫生—治疗学] R364[医药卫生—临床医学] R780.2