Screening And Validating Of Potential Key Genes In Regulating The Development Of Osteoarthritis

Background: Osteoarthritis(OA)is a very common degenerative disease of bone and joint,which is one of the main causes of disability in adults.The current pathological mechanism of OA is mainly defined as the change of articular cartilage.But the exact molecular mechanism is still unclear.Bioinformatics technology can help us to explore the internal mechanism of disease and create conditions for us to further understand and treat OA.Objective:The purpose of this study was to explore the potential key genes of OA and their interconnections in cartilage injury.Methods: The required OA gene database was obtained through GEO platform,and DEGs were obtained through processing and screening(screening condition:|log FC| ? 1.0 and adj.p.value <0.05).Then GO and KEGG enrichment analysis were carried out on the processed data through DAVID online website.The protein interaction network(PPI)of DEGs was constructed by Cytoscape program and the most important modules and pivotal genes were screened by the built-in program MCODE.Then,the main functions and related pathways of potential key genes were further analyzed,and the relationship of potential key genes in the destruction of OA cartilage was explored.Cartilage specimens of the fracture patients' femoral head and the knee joint of patients with OA in shenzhen second people's hospital were collected,and chondrocytes were isolated and cultured.Through the observation of cell morphology,toluidine blue staining experiment confirmed that it is indeed chondrocytes.The expression levels of COL I and COL II in chondrocytes were detected by immunofluorescence staining.The degree of chondrocyte inflammatory necrosis was detected by apoptosis staining.The expression levels of potential key genes in chondrocytes were analyzed by q RT-PCR.Results: In this study,265 DEGs were identified from the pathogenesis of OA by bioinformatics analysis and a protein network map was constructed.Then,4 important modules and 4 potential key genes were obtained through module analysis.Among the potential key genes,MAPK-14,PTPRC and PTPN12 were highly expressed in OA,while B9D1 was poorly expressed in OA.Experimental studies showed that the cytoplasm of the cultured cells was purplish blue after staining with toluidine blue,and the results were positive,indicating that the cells we cultured were chondrocytes.Compared with normal chondrocytes,OA chondrocytes had high expression of COL I protein and low expression of COL II protein.Moreover,the inflammatory necrosis of OA chondrocytes was significantly increased compared with that of normal chondrocytes,and the cell membrane was obviously destroyed.The differences in m RNA expression levels of four potential key genes in normal chondrocytes and OA chondrocytes were detected by q PCR.It was found that the expression levels of MAPK-14,PTPRC and PTPN12 in OA chondrocytes were significantly higher than that in normal chondrocytes,while the expression levels of B9D1 were significantly lower than that in normal chondrocytes.In addition,by looking up literature,MAPK-14 among the four potential key genes has been proved to play an important role in the treatment of OA.Conclusions:(1)Four potential key genes in the pathogenesis of OA were identified by bioinformatics screening,including MAPK-14,PTPRC,PTPN12 and B9D1;(2)The synthesis and catabolism of OA chondrocytes were unbalanced,and the destruction of cell membrane was increased.This may be related to the high expression of mapk-14 promoting the death of OA chondrocytes;(3)MAPK-14,PTPRC,PTPN12 and B9D1 are most closely related to the occurrence and development of OA.Regulating the expression of these key genes can provide potential reference targets for the treatment of osteoarthritis.