CD73通过AMPK/mTOR信号通路调控脊髓损伤机制研究  

Mechanism of CD73 regulates spinal cord injury of through AMPK/mTOR signaling pathway

作　　者：邵明昊[1] 靳明明 周萍[1] 郑超君[1] 朱巍[1] 马晓生[1] 吕飞舟[1] SHAO Ming-Hao;JIN Ming-Ming;ZHOU Ping;ZHENG Chao-Jun;ZHU Wei;MA Xiao-Sheng;LYU Fei-Zhou(Department of Spine Surgery,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院脊柱外科,上海200040 [2]协同中心影像平台,上海健康医学院,上海201318

出　　处：《中国免疫学杂志》2020年第19期2357-2360,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金青年项目(81802145);国家自然科学基金面上项目(81871552)。

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。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.

关 键 词：AMPK CD73 MTOR 脊髓损伤 

分 类 号：R310.34[医药卫生—基础医学]