骨髓间充质干细胞来源外泌体 lncRNA SNHG7 通过靶向 miR-485-5p/FSP1 轴减轻骨关节炎引起的软骨细胞铁死亡和炎症反应。
Exosomes-Shuttled lncRNA SNHG7 by Bone Marrow Mesenchymal Stem Cells Alleviates Osteoarthritis Through Targeting miR-485-5p/FSP1 Axis-Mediated Chondrocytes Ferroptosis and Inflammation.
机构信息
Rheumatology and Immunology Department, The First Hospital of Nanchang, No. 128, North Xiangshan Road, Nanchang, 330008, Jiangxi Province, China.
出版信息
Tissue Eng Regen Med. 2024 Dec;21(8):1203-1216. doi: 10.1007/s13770-024-00668-8. Epub 2024 Oct 3.
BACKGROUND
Osteoarthritis (OA), a degenerative joint disorder, is a major reason of disability in adults. Accumulating evidences have proved that bone marrow mesenchymal stem cells (BMSCs)-carried exosomes play a significant therapeutic effect on OA. However, the precise regulatory network remains unknown.
METHODS
OA and normal cartilage samples were acquired from patients, and chondrocytes were exposed to IL-1β to conduct a cellular OA model. Exosomes prepared from BMSCs were identified using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Cell viability was determined with CCK-8 assay. Inflammatory injury was assessed by LDH and inflammatory factors (TNF-α and IL-6) using corresponding ELISA kits, respectively. Ferroptosis was evaluated by GSH, MDA and iron levels using corresponding kits, and ROS level with DCFH-DA. The expressions of genes/proteins were determined with RT-qPCR/western bolt. RNA immunoprecipitation and luciferase activity assay were conducted for testing the interactions of small nucleolar RNA host gene 7 (SNHG7)/ferroptosis suppressor protein 1 (FSP1) and miR-485-5p.
RESULTS
The expressions of SNHG7 and FSP1 were both reduced in IL-1β-induced chondrocytes and OA cartilage tissues, and there was a positive correlation between them in clinical level. Moreover, SNHG7 was enriched in BMSCs-derived exosomes (BMSCs-Exos) and could be internalized by chondrocytes. Functional analysis illustrated that BMSCs-Exos administration repressed inflammatory injury, oxidative stress and ferroptosis in IL-1β-induced chondrocytes, while these changes were reinforced when SNHG7 was overexpressed in BMSCs-Exos. Notably, FSP1 silencing in chondrocytes abolished the beneficial effects mediated by exosomal SNHG7.
CONCLUSIONS
Exosomal SNHG7 released from BMSCs inhibited inflammation and ferroptosis in IL-1β-induced chondrocytes through miR-485-5p/FSP1 axis. This work suggested that BMSCs-derived exosomal SNHG7 would be a prospective target for OA treatment.
背景
骨关节炎（OA）是一种退行性关节疾病，是成年人残疾的主要原因。越来越多的证据证明骨髓间充质干细胞（BMSCs）来源的细胞外囊泡对 OA 具有显著的治疗作用。然而，确切的调控网络尚不清楚。
方法
从患者中获取 OA 和正常软骨样本，并将软骨细胞暴露于 IL-1β 中构建细胞 OA 模型。使用纳米颗粒跟踪分析（NTA）和透射电子显微镜（TEM）鉴定 BMSCs 来源的细胞外囊泡。通过 CCK-8 测定法测定细胞活力。分别使用相应的 ELISA 试剂盒测定 LDH 和炎性因子（TNF-α 和 IL-6）评估炎性损伤。使用相应试剂盒测定 GSH、MDA 和铁水平评估铁死亡，并用 DCFH-DA 测定 ROS 水平。通过 RT-qPCR/western bolt 测定基因/蛋白的表达。通过 RNA 免疫沉淀和荧光素酶活性测定进行小核仁 RNA 宿主基因 7（SNHG7）/铁死亡抑制蛋白 1（FSP1）和 miR-485-5p 相互作用的检测。
结果
在 IL-1β 诱导的软骨细胞和 OA 软骨组织中，SNHG7 和 FSP1 的表达均降低，且在临床水平上呈正相关。此外，SNHG7 在 BMSCs 来源的细胞外囊泡（BMSCs-Exos）中富集，并可被软骨细胞内化。功能分析表明，BMSCs-Exos 给药抑制了 IL-1β 诱导的软骨细胞的炎症损伤、氧化应激和铁死亡，而当 BMSCs-Exos 中 SNHG7 过表达时，这些变化得到增强。值得注意的是，在软骨细胞中沉默 FSP1 消除了外泌体 SNHG7 介导的有益作用。
结论
BMSCs 释放的细胞外囊泡 SNHG7 通过 miR-485-5p/FSP1 轴抑制 IL-1β 诱导的软骨细胞中的炎症和铁死亡。这项工作表明，BMSCs 来源的细胞外囊泡 SNHG7 可能成为 OA 治疗的一个有前景的靶点。
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