摘要
目的探讨低氧诱导的骨髓间充质干细胞(BMSCs)来源的外泌体对髓核细胞(NPCs)增殖及修复的影响。方法采用低氧环境(3%O2)对大鼠BMSCs进行预处理并提取外泌体。通过透射电子显微镜(TEM)、纳米颗粒追踪分析(NTA)和蛋白质免疫印迹法对外泌体进行鉴定。将NPCs与外泌体共培养,进行细胞增殖检测,分析细胞外基质的合成情况,并通过脂多糖建立炎性模型,检测肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的表达水平,通过实时荧光定量逆转录聚合酶链式反应(RT-qPCR)及免疫荧光分析低氧诱导的BMSCs来源的外泌体对炎性反应的影响。结果低氧诱导的BMSCs来源的外泌体可促进NPCs的增殖,并增强细胞外基质的修复能力,使Ⅰ型、Ⅲ型胶原和聚集蛋白聚糖(ACAN)的表达增加。低氧诱导的BMSCs来源的外泌体还可显著抑制炎性因子TNF-α和IL-1β的表达,减少NPCs的凋亡。结论低氧诱导的BMSCs来源的外泌体能够促进NPCs增殖、增强细胞外基质修复,并在炎性环境中减少NPCs凋亡,为其在椎间盘退行性变治疗中的潜在应用提供了理论基础。
Objective To investigate the effects of exosomes derived from bone marrow mesenchymal stem cells(BMSCs)induced by hypoxia on proliferation and repair of nucleus pulposus cells(NPCs).Methods Rat BMSCs were pretreated in hypoxic environmen(t 3%O2),and exosomes were extracted.Exosomes were identified by transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA)and Western bloting.NPCs were co-cultured with exosomes for cell proliferation detection,the synthesis of extracellular matrix was analyzed,and an inflammatory model was established through lipopolysaccharide to detect the expression levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β),and effect of exosomes secreted by hypoxic BMSCs on the inflammatory response was analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)and immunofluorescence.Results Exosomes derived from BMSCs induced by hypoxia promoted the proliferation of NPCs and enhanced the repair ability of extracellular matrix,and increased the expression of CollagenⅠ,CollagenⅢand aggrecan(ACAN).Exosomes derived from BMSCs induced by hypoxia also significantly inhibited the expression of inflammatory factors TNF-α and IL-1β,reduced the apoptosis of NPCs.Conclusion Exosomes derived from BMSCs induced by hypoxia can promote the proliferation of NPCs,enhance extracellular matrix repair,and reduce the apoptosis of NPCs in inflammatory environment,providing a theoretical basis for its potential application in the treatment of intervertebral disc degeneration.
作者
袁佳滨
宫峰
罗毓钦
石志才
谷晓川
Yuan Jiabin;Gong Feng;Luo Yuqin;Shi Zhicai;Gu Xiaochuan(Department of Orthopaedics,Changhai Hospital,Naval Medical University,Shanghai 200433,China;Department of Plastic Surgery,Changhai Hospital,Naval Medical University,Shanghai 200433,China)
出处
《脊柱外科杂志》
2025年第4期233-238,259,共7页
Journal of Spinal Surgery
关键词
间质干细胞
旁分泌细胞细胞间通路
细胞增殖
细胞分化
椎间盘退行性变
Mesenchymal stem cells
Paracrine communication
Cell proliferation
Cell differentiation
Intervertebral disc degeneration
