长链非编码RNA NEAT1对神经胶质细胞功能的调控在SLE狼疮脑病发病中的作用及机制研究

Neuropsychiatric SLE (NPSLE) is a fatal complication of SLE patients and difficult to early diagnosis, the pathogenesis of NPSLE is not clear. The inflammatory chemokines such as IL-6, etc. were found abnormal elevated in cerebrospinal fluid of NPSLE patients, which may play an important role in the cause of NPSLE. It has been reported that over expression of lncRNA-NEAT1 in peripheral blood mononuclear cells of SLE patients can activate MAPK pathway to promote the secretion of cytokines IL-6 and CXCL10. The gial cells have been proven to participate in the course of encephalitis in the same way exactly. The latest study showed that the excitability of iPSC cells was closely related to the expression of lncRNA-NEAT1, suggesting that lncRNA-NEAT1 may regulate the function of glial cells in the pathogenesis of NPSLE. We launch this research to investigate the role of lncRNA NEAT1 in the regulation of glial cells and its related mechanism in the early stage of the disease with the help of functional magnetic resonance imaging, as well as to set the foundation of the early diagnosis and treatment of NPSLE.

狼疮脑病(NPSLE)是直接威胁系统性红斑狼疮患者生命的严重并发症且难以早期诊断，其发病机制尚不明确，NPSLE患者脑脊液中异常升高的炎症趋化因子如IL-6等可能在其中发挥重要作用。研究发现在系统性红斑狼疮患者的外周血单个核细胞中过表达lncRNA-NEAT1可激活MAPK通路进而促进细胞因子IL-6、CXCL10的分泌，而神经胶质细胞亦可通过相同途径参与中枢免疫调节，最新研究表明iPSC神经细胞兴奋性与lncRNA-NEAT1的表达水平密切相关，提示lncRNA-NEAT1可能通过调节神经胶质细胞的功能而在狼疮脑病的发病机制中起到重要作用。本申请项目拟在前期研究的基础上结合神经系统磁共振结构功能成像技术深入探讨lncRNA NEAT1对狼疮脑病神经胶质细胞的调控作用及其相关机制，为开展狼疮脑病的早期诊断和治疗药物的开发奠定基础。

本研究以探索SLE患者狼疮脑病发病与神经胶质细胞的相关性为核心，应用多模态MRI成像方法（DTI、ASL、MRS）探索SLE患者脑组织病理性改变，定位早期病变部位及细胞类型（病灶数量、部位、WMHIs、GCA、MTA、Scheltens评分等），总结具有高敏感性和特异性的狼疮脑病影像学标志物，本研究发现SLE患者胼胝体、额叶白质区DTI序列FA值较对照组降低；灌注成像分析示SLE患者双侧岛叶区域的CBF值相比正常对照组降低，提示其为狼疮脑病的早期表现，对临床诊断与患者预后都具有极大意义。同时建立狼疮脑模型鼠，通过影像学及病理学验证狼疮脑病的发生发展与神经胶质细胞功能的关联性，发现狼疮脑病组较对照组海马区功能学成像波普NAA峰降低，Cho峰升高，提示狼疮脑病组神经系统存在相应区域的神经纤维破坏和胶质细胞增生；小鼠脑组织免疫荧光双标MBP（髓鞘结合蛋白）及DAPI（胞核）示狼疮脑病组MBP的荧光强度低于对照组，验证了狼疮脑病组脑白质纤维受到破坏。在此基础上探索神经胶质细胞细胞株（BV-22）分泌细胞因子功能是否与LncRNA NEAT1表达相关，分别利用shRNA敲低Lnc RNA NEAT1/构建LncRNA NEAT1过表达病毒，并通过检测BV-2分泌细胞因子来反应LncRNA NEAT1对神经胶质细胞活化水平的影响，发现BV-2细胞株分泌细胞因子水平与LncRNA NEAT1正相关，证实LncRNA NEAT1具有一定调节BV-2分泌细胞因子的功能，结合既往发现狼疮脑病病人PBMC中LncRNA NEAT1升高及狼疮脑患者脑脊液中细胞因子水平显著高于正常人的研究结论，推测狼疮患者LncRNA NEAT1异常表达可能也作用于神经胶质细胞进而导致狼疮脑病的发生和发展。综上，本研究临床方面为建立基于多模态神经影像学的狼疮脑病人工智能诊断模型夯实基础，有利于提高狼疮脑病的早期诊断的敏感性；发病机制方面以LncRNA是否参与调控神经胶质细胞分泌功能为切入点，探讨LncRNA NEAT1对BV-2细胞炎症因子分泌的调控及具体机制并得出初步结论，在国内外尚未见相关报道，为探寻狼疮脑病的精准治疗靶点提供理论依据。课题组已发表标注本课题资助的论文2篇，另有3篇英文论文在投，并成功申请实用新型专利1项。