【摘要】 目的:本研究旨在探讨CD73在脊髓损伤中的调控机制。方法:采用CD73基因敲除型(KO)和野生型(WT)C57BL/6小鼠建立脊髓损伤(SCI)模型。采用HE染色、Nissl染色和IHC染色分析CD73、IL-1β、TNF-α和Caspase-3的表达以评估脊髓的病理改变,TUNEL染色检测神经元凋亡;qPCR检测CD73、IL-6、IL-1β、TNF-α和Caspase-3的mRNA水平,RNA测序鉴定差异表达基因。结果:SCI组IL-6、IL-1β、TNF-α、Caspase-3 mRNA表达较正常组明显升高。CD73 KO小鼠神经元凋亡增多,AMPK明显上调, mTOR受到抑制。RNA测序结果显示,4Ebp1在WT-SCI小鼠中显著上调。Western blot结果显示WT组小鼠4Epb1较KO小鼠组明显升高。结论:在SCI小鼠模型中,CD73能够抑制脊髓继发性损伤。下调CD73可通过调节AMPK/mTOR信号通路诱导更严重的神经元凋亡。CD73可能是SCI治疗靶点之一。更多还原

【Abstract】 Objective:Purpose of this study was to explore the regulatory mechanism of CD73 in spinal cord injury.Methods:Spinal cord injury(SCI) models were established using CD73 gene knockout(KO) and wild type(WT) C57 BL/6 mice.HE staining,Nissl staining,and IHC staining were used to evaluate expressions of CD73,IL-1β,TNF-α,and Caspase-3 to assess pathologic changes in spine.TUNEL staining was used to detect neuronal apoptosis;qPCR was used to evaluate mRNA of CD73,IL-6,IL-1β,TNF-α and Caspase-3.RNA sequencing was used to identify differentially expressed genes.Results:Expressions of IL-6,IL-1β,TNF-α,and Caspase-3 mRNA in SCI group were significantly higher than those in normal group.CD73 KO mice had increased neuronal apoptosis,significantly upregulated AMPK,and inhibited mTOR.Results of RNA sequencing showed that 4 Ebp1 was significantly up-regulated in WT-SCI mice.Western blot showed that 4 Epb1 in WT mice was significantly higher than that in KO mice.Conclusion:In SCI mice model,CD73 can inhibit secondary spinal cord injury.Down-regulation of CD73 can induce more severe neuronal apoptosis by regulating the AMPK/mTOR signaling pathway.CD73 may be one of the targets for SCI treatment.更多还原