Department of orthopedics, Jincheng People's Hospital, China.
Department of orthopedics, Jincheng People's Hospital, China.
Tissue Cell. 2023 Jun;82:102046. doi: 10.1016/j.tice.2023.102046. Epub 2023 Feb 21.
Osteoarthritis is a main cause of deformity in aging people. The chondrogenesis of human adipose-derived stem cells (hADSCs) has a positive effect on the cure of osteoarthritis. However, the regulatory mechanism of hADSC chondrogenesis still needs further exploration. This research investigates the role of interferon regulatory factor 1 (IRF1) in the chondrogenesis of hADSCs.
hADSCs were purchased and cultured. The interaction between IRF1 and hypoxia inducible lipid droplet associated (HILPDA) was predicted by bioinformatics analysis, and verified through dual-luciferase reporter and chromatin immunoprecipitation assays. The expressions of IRF1 and HILPDA in osteoarthritis cartilage samples were measured through qRT-PCR. After hADSCs were transfected or further induced for chondrogenesis, the chondrogenesis was visualized by Alcian blue staining, and the expressions of IRF1, HILPDA and chondrogenesis-related factors (SOX9, Aggrecan, COL2A1, MMP13, MMP3) were determined through qRT-PCR or Western blot.
HILPDA bound to IRF1 in hADSCs. IRF1 and HILPDA levels were up-regulated during the chondrogenesis of hADSCs. Overexpressions of IRF1 and HILPDA promoted the chondrogenesis of hADSCs with the up-regulation of SOX9, Aggrecan and COL2A1 and the down-regulation of MMP13 and MMP3; however, IRF1 silencing generated the opposite effects. Besides, HILPDA overexpression reversed the effects of IRF1 silencing on inhibiting chondrogenesis of hADSCs and regulating the expressions of chondrogenesis-related factors.
IRF1 promotes the chondrogenesis of hADSCs through up-regulating HILPDA level, providing novel biomarkers for treating osteoarthritis.
骨关节炎是老年人畸形的主要原因。人脂肪来源干细胞（hADSCs）的软骨生成对骨关节炎的治疗有积极作用。然而，hADSC 软骨生成的调节机制仍需要进一步探索。本研究探讨干扰素调节因子 1（IRF1）在 hADSC 软骨生成中的作用。
购买并培养 hADSCs。通过生物信息学分析预测 IRF1 与缺氧诱导的脂滴相关（HILPDA）之间的相互作用，并通过双荧光素酶报告和染色质免疫沉淀检测进行验证。通过 qRT-PCR 测量骨关节炎软骨样本中 IRF1 和 HILPDA 的表达。在 hADSCs 转染或进一步诱导软骨生成后，通过 Alcian 蓝染色观察软骨生成，通过 qRT-PCR 或 Western blot 测定 IRF1、HILPDA 和软骨生成相关因子（SOX9、聚集蛋白聚糖、COL2A1、MMP13、MMP3）的表达。
HILPDA 在 hADSCs 中与 IRF1 结合。IRF1 和 HILPDA 水平在 hADSCs 的软骨生成过程中上调。IRF1 和 HILPDA 的过表达促进 hADSCs 的软骨生成，同时上调 SOX9、聚集蛋白聚糖和 COL2A1，下调 MMP13 和 MMP3；然而，IRF1 沉默则产生相反的效果。此外，HILPDA 的过表达逆转了 IRF1 沉默对抑制 hADSCs 软骨生成和调节软骨生成相关因子表达的作用。
IRF1 通过上调 HILPDA 水平促进 hADSCs 的软骨生成，为治疗骨关节炎提供了新的生物标志物。
