Bioinformatic analysis and experimental validation of ferroptosis in osteoarthritis

doi:10.12307/2023.437

Abstract

Abstract: BACKGROUND: Regulating ferroptosis by targeting iron overload genes may be a fast and effective way to delay the degeneration of osteoarthritis. However, the molecular mechanisms and gene targets related to ferroptosis in osteoarthritis are still unclear.
OBJECTIVE: To analyze the key genes and pathways of ferroptosis in osteoarthritis by bioinformatics analysis, verify the marker genes of ferroptosis in combination with in vitro experiments, and explore the potential role of ferroptosis in osteoarthritis.
METHODS: “Osteoarthritis” was used as a keyword. Relevant public data from the GEO database between January 1, 2010 and January 1, 2021 were retrieved. A gene microarray dataset GSE55235 was yielded and differentially expressed genes were identified after data correction analysis of the GSE55235 data set. Ferroptosis-related genes retrieved by the FerrDb database were intersected with differentially expressed genes in the GSE55235 dataset, and th intersected genes were used for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis and the protein-protein interaction network was mapped. Ferroptosis-related HUB genes in osteoarthritis were obtained and ferroptosis marker genes were then selected. Human chondrocytes were divided into control group (normal human chondrocytes) and experimental group (cellular osteoarthritic model). The mRNA expression of ferroptosis marker genes in the two groups was detected by quantitative real-time fluorescence PCR.
RESULTS AND CONCLUSION: A total of 36 ferroptosis-related genes in osteoarthritis were obtained. Gene Ontology enrichment analysis showed that these differentially expressed genes were mainly involved in oxidative stress, corticosteroids response, steroid hormone response, glucocorticoid response, and reactive oxygen metabolism, and play a role in NAD(P)H oxidase and oxidoreductase activity, heme binding, and tetrapyrrole binding that produce superoxides. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that ferroptosis-related genes in osteoarthritis were mainly involved in NOD-like receptor, interleukin-17, hypoxia-inducible factor-1, tumor necrosis factor, forkhead box transcription factor class O. After constructing the protein-protein interaction network, ferroptosis-related HUB genes, including vascular endothelial growth factor A, interleukin-6, JUN, PTGS2, DUSP1, were identified. Four ferroptosis marker genes, including vascular endothelial growth factor A, interleukin-6, PTGS2, and DUSP1, were selected. In vitro experiments confirmed significant differences between the control and experimental groups (P < 0.05). The overall findings indicate that ferroptosis may act on osteoarthritis through oxidative stress, redox and induced inflammation, which may be related to NAD(P) H stimulation by signaling pathway such as hypoxia-inducible factor 1. Vascular endothelial growth factor A, interleukin-6, PTGS2, and DUSP1 can be used as biomarkers of ferroptosis in osteoarthritis.

Key words: osteoarthritis, ferroptosis, gene, signaling pathway, bioinformatics, chondrocyte, cellular experiment, biomarker, oxidative stress, inflammation

CLC Number:

Li Zhe, Yuan Changshen, Guan Yanbing, Xu Wenfei, Liao Shuning, Rong Weiming, Mei Qijie, Duan Kan. Bioinformatic analysis and experimental validation of ferroptosis in osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(17): 2637-2643.

Figures/Tables 9

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