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2022, Vol. 26 ›› Issue (11): 1680-1685Quantitative analysis of transcriptome and proteome of knee joint synovial cells with osteopontin knockdownXu Renjie1, 2, Zhong Qiao1, Liu Yubo1, Yu Xiao1, Yan Yongqing3, Saijilafu2, Yang Huilin2, Chen Guangxiang1
Abstract: BACKGROUND: The mRNA and protein levels of osteopontin in articular cartilage and synovial fluid of patients with osteoarthritis are significantly higher than those of normal controls. However, the role and specific mechanism of osteopontin in the development of knee osteoarthritis have not been clarified. OBJECTIVE: To investigate the level of osteopontin in synovial cells of knee joint osteoarthritis patients and its relationship with the severity of osteoarthritis, and to further elucidate the mechanism that affects the progression of osteoarthritis through transcriptome and proteome sequencing. METHODS: A total of 42 patients with primary knee osteoarthritis diagnosed and treated in 2019 were divided into unicompartment knee osteoarthritis group and bicompartment knee osteoarthritis group based on the involvement of the tibiofemoral joint. Synovial and joint fluid specimens were harvested. PCR method was used to measure the abundance of osteopontin gene expression in synovial specimens, and ELISA method was used to measure osteopontin protein level in synovial fluid specimens. Enzymatic digestion method was used to obtain and culture human knee osteoarthritis synovial fibroblast-like cells. After osteopontin-siRNA intervention for 24 hours, the samples were tested for transcriptome and proteome. RESULTS AND CONCLUSION: Compared with the unicompartment knee osteoarthritis group, the osteopontin levels in the synovium and synovial fluid of the bicompartment knee osteoarthritis group increased significantly (P < 0.05). In in vitro cultured human knee osteoarthritis synovial fibroblast-like cells, after osteopontin knockdown by siRNA, quantitative studies of the transcriptome and proteome identified 14 662 transcripts and 3 608 proteins, respectively. Among them, the transcriptome and proteome analysis results of five proteins showed a simultaneous decrease, and the transcriptome and proteome analysis results of two proteins showed a simultaneous increase. To conclude, osteopontin has an adverse effect on chondrocytes. The mechanism may be through the pro-inflammatory effect and the negative effect of insulin-like growth factor-binding protein 3 on cartilage. Key words:knee joint, osteoarthritis, osteopontin, synovial cells, transcriptome, proteome, mechanism |