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Phosphorylation of osteopontin has proapoptotic and proinflammatory effects on human knee osteoarthritis chondrocytes

机译:骨桥蛋白的磷酸化对人膝骨关节炎软骨细胞具有促凋亡和促炎作用

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摘要

The aim of the present study was to investigate the effects of phosphorylated osteopontin (p-OPN) on apoptosis and pro-inflammatory cytokine expression in human knee osteoarthritis (OA) chondrocytes. Human knee OA chondrocytes obtained from patients who underwent total knee arthroplasty were treated with p-OPN, OPN or buffer. Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) and western blot analysis were used to assess the expression levels of proinflammatory factors, including interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6 and nuclear factor (NF)-κB. Apoptosis of human knee OA chondrocytes was detected by Annexin V-fluorescein isothiocyanate/propidium iodide flow cytometry. Compared with the controls, chondrocytes treated with OPN exhibited higher mRNA and protein expression levels of proinflammatory factors (IL-1β, TNF-α, IL-6 and NF-κB), and a higher percentage of apoptotic chondrocytes. Furthermore, chondrocytes treated with p-OPN exhibited the highest mRNA and protein expression levels of proinflammatory factors (IL-1β, TNF-α, IL-6, NF-κB) and the highest percentage of apoptotic chondrocytes. p-OPN induces chondrocyte apoptosis and proinflammatory factor release, which suggests that p-OPN may contribute to OA pathogenesis, and inhibition of p-OPN may provide a novel effective strategy to slow or halt OA progression.
机译:本研究的目的是研究磷酸化骨桥蛋白(p-OPN)对人膝骨关节炎(OA)软骨细胞凋亡和促炎性细胞因子表达的影响。对接受全膝关节置换术的患者的人膝OA软骨细胞进行p-OPN,OPN或缓冲液处理。用逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹分析来评估促炎因子的表达水平,包括白介素(IL)-1β,肿瘤坏死因子(TNF)-α,IL-6和核因子(NF)-κB。通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶流式细胞术检测人膝OA软骨细胞的凋亡。与对照组相比,用OPN处理的软骨细胞表现出较高的促炎因子(IL-1β,TNF-α,IL-6和NF-κB)mRNA和蛋白表达水平,并具有较高百分比的凋亡软骨细胞。此外,经p-OPN处理的软骨细胞表现出促炎因子(IL-1β,TNF-α,IL-6,NF-κB)的最高mRNA和蛋白质表达水平,以及最高百分比的凋亡软骨细胞。 p-OPN诱导软骨细胞凋亡和促炎因子释放,这表明p-OPN可能导致OA发病机理,而p-OPN的抑制则可能提供一种新的有效策略来减缓或阻止OA的发展。

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