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Abstract
Background
Osteoarthritis (OA) is a common joint disease characterized by progressive cartilage degradation. Circular RNAs (circRNAs) are involved in the initiation and development of OA. This study aimed to explore the potential role and mechanism of circRNA protein kinase C eta (circ-PRKCH) in OA.
Methods
A total of 30 cartilage specimens were collected from OA patients or normal subjects. Human chondrocytes (CHON-001) were stimulated with interleukin-1β (IL-1β) to establish an in vitro OA model. The expression levels of circ-PRKCH, microRNA-502-5p (miR-502-5p) and circ-PRKCH or A disintegrin and metalloproteases metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) in cartilage specimens and IL-1β-treated chondrocytes were detected by quantitative real-time PCR or Western blot, and their correlation in OA cartilage specimens was analysed by Spearman’s correlation coefficient. The targeted relationship between miR-502-5p and circ-PRKCH or ADAMTS5 was verified by dual-luciferase reporter assay and RNA Immunoprecipitation (RIP) assay. Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2′-deoxyuridine (EDU), flow cytometry, wound healing and enzyme-linked immunosorbent assay (ELISA) assays were applied to evaluate cell proliferation, apoptosis, migration and inflammatory response in IL-1β-treated chondrocytes. Exosomes were identified by transmission electron microscope (TEM) and Western blot.
Results
Circ-PRKCH and ADAMTS5 expression levels were up-regulated, while miR-502-5p expression was down-regulated in OA cartilage tissues and IL-1β-treated chondrocytes. Depletion of circ-PRKCH relieved IL-1β-treated chondrocyte cell phenotypic changes by promoting cell proliferation and migration, as well as inhibiting apoptosis and inflammatory response. Mechanically, circ-PRKCH acted as a sponge for miR-502-5p to regulate ADAMTS5 expression, thereby contributing to IL-1β-treated chondrocyte cell phenotypic changes. Moreover, exosomes derived from IL-1β-treated chondrocytes could transfer circ-PRKCH across cells.
Conclusion
Circ-PRKCH contributed to IL-1β-treated cell phenotypic changes in chondrocytes via modulating miR-502-5p/ADAMTS5 pathway, which might provide a promising biomarker for OA treatment.
Ethical approval
The research related to human use has been complied with all the relevant national regulations, and has been approved by the Ethics Committee of Affiliated Hospital of Chifeng University
Author contributions
Zhongxing Liu and Jian Cao participated in the design of the work, methodology, data interpretation and drafted the manuscript. Limin Zhang and Jinlong Li participated in the collection of data and analysis for the work, carried out the statistical analysis. Tinghan Yan participated in data interpretation and methodology. Peng Zhou interpreted the data, provided Resourse and Data curation. Sidi Zhang participated in the methodology and Formal Analysis.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Data availability statement
Data analysed for this study will be available on a reasonable request.