目的:骨关节炎(OA)的发病机制尚不清楚,导致缺乏针对性的治疗。我们旨在探讨载脂蛋白D(APOD)的作用,我们之前通过生物信息学分析研究的关键基因,在体外成纤维细胞样滑膜细胞(FLS)和软骨细胞上,确认其对OA进展延迟的重要作用。
方法:从OA患者的滑膜和软骨中提取原代FLS和软骨细胞,体外用白细胞介素1β(IL-1β)刺激。APOD干预后,检测FLS和软骨细胞的活力和增殖。随后,qRT-PCR检测两种细胞的炎症因子,ELISA和Westernblot,同时测定细胞凋亡和自噬相关物质。最后,检测FLS和软骨细胞的氧化水平。
结果:APOD逆转了IL-1β刺激的FLS和软骨细胞中基因表达的变化。APOD减轻FLS的增殖,促进软骨细胞的增殖,降低炎症因子的表达。此外,APOD促进FLS细胞凋亡和软骨细胞自成像,同时减少软骨细胞的凋亡。最后,APOD干预后观察到两个细胞的活性氧(ROS)水平降低,以及抗氧化相关基因的表达增加。
结论:APOD对FLS和软骨细胞的增殖有影响,可能通过细胞凋亡、自体照相术以及降低氧化应激,延缓OA的进展。本文受版权保护。保留所有权利。
The pathogenesis of osteoarthritis (OA) is still unclear, leading to the lack of targeted treatment. We aimed to probe into the effect of apolipoprotein D (
APOD), the key gene from our previous study through bioinformatics analysis, on fibroblast-like synoviocyte (FLS) and chondrocytes in vitro to confirm its potential roles on the delay of OA progression. Primary FLS and chondrocytes were extracted from synovium and cartilage of OA patients and stimulated with interleukin 1β (IL-1β) in vitro. After
APOD intervention, viability and proliferation of FLS and chondrocytes were detected. Subsequently, the inflammatory factors of the two cells were detected by quantitative reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and western blot, and the apoptosis and autophagy-related substances were determined at the same time. Finally, the oxidation level in FLS and chondrocytes were detected.
APOD reversed the change of gene expression stimulated by IL-1β in FLS and chondrocytes.
APOD alleviated the proliferation of FLS while promoted proliferation of chondrocytes, and reduced the expression of inflammatory factors. Moreover, APOD promoted apoptosis of FLS and autography of chondrocytes, while reduced apoptosis of chondrocytes. Finally, decrease level of reactive oxygen species (ROS) in both cells were observed after APOD intervention, as well as the increased expression of antioxidant-related genes.
APOD had effects on the proliferation of FLS and chondrocytes through apoptosis and autography as well as the reduction of oxidative stress, delaying the progress of OA.