Xiao Bin, Wang Guozhu, Huo Hongliang, Li Weiwei
Department of Orthopedics, Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine, Xianyang, Shaanxi 712000, P.R. China.
Exp Ther Med. 2021 Oct;22(4):1115. doi: 10.3892/etm.2021.10549. Epub 2021 Aug 4.
Kashin-Beck disease (KBD) is a chronic and endemic osteoarthropathy. The pathogenesis of KBD has yet to be fully elucidated, although previous studies have shown that its etiology may be associated with low selenium abundance and high exposure to mycotoxins, such as T-2 toxin. In the present study, the Comparative Toxicogenomics Database was used to identify key genes associated with KBD, T-2 toxin and selenium. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to identify the biological processes and pathways that key genes may be associated with. By searching the Search Tool for the Retrieval of Interacting Genes database and the Molecular Complex Detection plug-in with Cytoscape, it was possible to construct a KBD-associated protein-protein interaction (PPI) network, and screen the core modules and genes. Western blot analysis was subsequently used to verify the expression levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor A (VEGFA), two components that are associated with the HIF-1 signaling pathway in KBD disease. Via this approach, a total of 301 key genes were identified that were associated with KBD, T-2 toxin and selenium. The results of the GO and KEGG enrichment analyses demonstrated that these key genes were mainly involved in the process of apoptosis. Previous studies have demonstrated that excessive apoptosis of chondrocytes plays a crucial role in the pathophysiology of KBD, and that HIF-1α has an important role in chondrocyte apoptosis; therefore, the present study was focused on the expression level of HIF-1α in KBD. By analyzing the PPI network constructed from the key genes, a total of 10 core genes were obtained that may be associated with KBD. The results of western blotting experiments revealed that, after treating chondrocytes with different concentrations of T-2 toxin, the expression levels of HIF-1α and VEGFA were markedly downregulated. The iRegulon plug-in for Cytoscape was used to predict the transcription factors that may regulate HIF-1α and VEGFA in the HIF-1 signaling pathway. Using this approach, 10 core genes and 15 transcription factors were obtained. These results may help to clarify the pathogenesis of KBD, thereby providing further avenues for the therapeutic treatment of KBD.
大骨节病（KBD）是一种慢性地方性骨关节病。尽管先前的研究表明其病因可能与低硒含量和高暴露于霉菌毒素（如T-2毒素）有关，但大骨节病的发病机制尚未完全阐明。在本研究中，利用比较毒理基因组学数据库来鉴定与大骨节病、T-2毒素和硒相关的关键基因。采用基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析来鉴定关键基因可能涉及的生物学过程和途径。通过搜索检索相互作用基因数据库的搜索工具和Cytoscape的分子复合物检测插件，构建了与大骨节病相关的蛋白质-蛋白质相互作用（PPI）网络，并筛选出核心模块和基因。随后采用蛋白质印迹分析来验证缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子A（VEGFA）的表达水平，这两种成分与大骨节病中的HIF-1信号通路相关。通过这种方法，共鉴定出301个与大骨节病、T-2毒素和硒相关的关键基因。GO和KEGG富集分析结果表明，这些关键基因主要参与细胞凋亡过程。先前的研究表明，软骨细胞的过度凋亡在大骨节病的病理生理学中起关键作用，并且HIF-1α在软骨细胞凋亡中起重要作用；因此，本研究聚焦于大骨节病中HIF-1α的表达水平。通过分析由关键基因构建的PPI网络，共获得10个可能与大骨节病相关的核心基因。蛋白质印迹实验结果显示，用不同浓度的T-2毒素处理软骨细胞后，HIF-1α和VEGFA的表达水平显著下调。利用Cytoscape的iRegulon插件预测在HIF-1信号通路中可能调节HIF-1α和VEGFA的转录因子。通过这种方法，获得了10个核心基因和15个转录因子。这些结果可能有助于阐明大骨节病的发病机制，从而为大骨节病的治疗提供进一步的途径。
