通过综合生物信息学分析和实验验证鉴定与骨科植入物维生素E混合超高分子量聚乙烯碎片相关的炎症性骨溶解关键候选基因

Identification of Key Candidate Genes Related to Inflammatory Osteolysis Associated with Vitamin E-Blended UHMWPE Debris of Orthopedic Implants by Integrated Bioinformatics Analysis and Experimental Confirmation.

作者信息

Liu Fanxiao, Dong Jun, Zhou Dongsheng, Zhang Qingyu

机构信息

Department of Orthopaedics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People's Republic of China.

出版信息

J Inflamm Res. 2021 Jul 26;14:3537-3554. doi: 10.2147/JIR.S320839. eCollection 2021.

DOI:10.2147/JIR.S320839

PMID:34345178

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8323865/

Abstract

PURPOSE

This study aims to identify differentially expressed genes (DEGs) in macrophages exposed to ultra-high-molecular-weight polyethylene (UHMWPE) or vitamin E-blended UHMWPE (VE-UHMWPE) particles, thereby providing potential targets for the treatment of inflammatory osteolysis.

METHODS

The GSE104589 dataset of genome expression in macrophages exposed to UHMWPE and VE-UHMWPE was downloaded from the Gene Expression Omnibus database to identify DEGs. Functional enrichment analysis was performed using DAVID, and the corresponding protein-protein interaction (PPI) network was constructed from the STRING database. Important modules were selected using the molecular complex detection algorithm, and hub genes were identified in cytoHubba. MicroRNAs targeting these DEGs were obtained from the TarBase, miRTarBase, and miRecords databases, while transcription factors (TFs) targeting DEGs were predicted from the ENCODE database. Finally, the top five DEGs were validated by quantitative real-time polymerase chain reaction (qRT-PCR).

RESULTS

A total of 112 DEGs (44 upregulated and 68 downregulated DEGs) were screened. Immune and inflammatory responses were significantly related in gene ontology analysis, and 18 signaling pathways were enriched according to Kyoto Encyclopedia of Genes and Genomes pathway analysis. The PPI network involving 85 nodes and 266 protein pairs indicated that IL1β, CXCL1, ICAM1, CCL5 and CCL4 showed higher degrees. qRT-PCR analysis of the top five DEGs revealed a decreasing trend in the VE-UHMWPE group compared with the UHMWPE group. Key microRNAs (hsa-miR-144, hsa-miR-21, and hsa-miR-221) and TFs (RELA and NFKB1) were predicted to be correlated with the pathogenesis of inflammatory osteolysis through microRNA-TF regulatory network analysis.

CONCLUSION

The present study helps shed light on the molecular mechanisms underlying the changes in the wear-induced inflammatory process after blending vitamin E with UHMWPE. Hub genes including IL1β, CXCL1, ICAM1, CCL5, and CCL4, key microRNAs (hsa-miR-144, hsa-miR-21, and hsa-miR-221) and TFs (RELA and NFKB1) may serve as prognostic and therapeutic targets of inflammatory osteolysis.

摘要

目的

本研究旨在鉴定暴露于超高分子量聚乙烯（UHMWPE）或维生素E混合超高分子量聚乙烯（VE-UHMWPE）颗粒的巨噬细胞中差异表达基因（DEG），从而为炎性骨溶解的治疗提供潜在靶点。

方法

从基因表达综合数据库下载暴露于UHMWPE和VE-UHMWPE的巨噬细胞基因组表达的GSE104589数据集，以鉴定DEG。使用DAVID进行功能富集分析，并从STRING数据库构建相应的蛋白质-蛋白质相互作用（PPI）网络。使用分子复合物检测算法选择重要模块，并在cytoHubba中鉴定枢纽基因。从TarBase、miRTarBase和miRecords数据库中获取靶向这些DEG的微小RNA，同时从ENCODE数据库预测靶向DEG的转录因子（TF）。最后，通过定量实时聚合酶链反应（qRT-PCR）验证前五个DEG。

结果

共筛选出112个DEG（44个上调DEG和68个下调DEG）。基因本体分析显示免疫和炎症反应显著相关，根据京都基因与基因组百科全书通路分析，有18条信号通路富集。涉及85个节点和266个蛋白质对的PPI网络表明，IL1β、CXCL1、ICAM1、CCL5和CCL4具有较高的度数。对前五个DEG的qRT-PCR分析显示，与UHMWPE组相比，VE-UHMWPE组呈下降趋势。通过微小RNA-TF调控网络分析预测关键微小RNA（hsa-miR-144、hsa-miR-21和hsa-miR-221）和TF（RELA和NFKB1）与炎性骨溶解的发病机制相关。

结论

本研究有助于阐明维生素E与UHMWPE混合后磨损诱导的炎症过程变化的分子机制。包括IL1β、CXCL1、ICAM1、CCL5和CCL4在内的枢纽基因、关键微小RNA（hsa-miR-144、hsa-miR-21和hsa-miR-221）和TF（RELA和NFKB1）可能作为炎性骨溶解的预后和治疗靶点。