甲强龙抑制脊髓损伤后内源性神经干细胞增殖中LncRNA-NRA的关键调控作用及机制研究

Spinal cord injury (SCI) is a serious central nervous system trauma that causes disability. Although methylprednisolone (MP) can relieve secondary SCI, the neuroprotective effection is counteracted by the side effects that inhibit eNSCs’ proliferation and neurorestoration, which leading MP does not improve the long-term neural function. Our research group has confirmed that MP can inhibit the proliferation of stem cells by down-regulating the Notch pathway, but the pathway is not the key target to reverse the side effects. Further research found that MP can up-regulate LncRNA-NRA, then down-regulate Notch receptor 1 expression and Notch pathway in eNSCs, and ultimately inhibit eNSCs proliferation. LncRNA-NRA may be a key regulator in MP inhibits eNSCs proliferation, but further confirmation is needed, and the mechanism of LncRNA-NRA down-regulating the Notch pathway is unknown. We propose to study the mechanism of LncRNA-NRA regulates Notch Receptor-1 expression and Notch pathway, and key regulatory role of LncRNA-NRA in MP inhibits eNSCs proliferation, and provide an effective antagonistic targets to reverse the MP inhibits eNSCs proliferation with the experimental technique such as Gene chips, RNA pull-down, RIP, Protein mass spectrometry analysis, Biological information analysis etc. The study will provide a new theoretical basis for the application of MP in acute spinal cord injury.

脊髓损伤是严重的中枢神经系统创伤。过去认为大剂量甲强龙冲击可减轻急性期继发性脊髓损伤，但新近研究证实其治疗效果有限，不能改善远期神经功能。我们前期证实甲强龙通过下调内源性神经干细胞（eNSCs）中Notch受体1（NR1）表达，从而抑制eNSCs增殖，最终阻碍脊髓损伤后神经修复，但甲强龙作用eNSCs后导致细胞NR1表达下调的具体机制不清。预实验结果发现甲强龙可上调eNSCs中长链非编码RNA-NRA（LncRNA-NRA）表达，这是导致NR1表达下调、抑制eNSCs增殖的关键所在。我们拟通过基因芯片、RNA pull-down、RIP、生物学信息分析等手段明确LncRNA-NRA调控NR1表达的具体机制，及其在甲强龙抑制eNSCs增殖中的关键调控作用，为解决甲强龙在急性脊髓损伤中应用瓶颈提供新的理论依据。

脊髓损伤是严重的中枢神经系统创伤。过去认为大剂量甲强龙冲击可减轻急性期继发性脊.髓损伤，但新近研究证实其治疗效果有限，不能改善远期神经功能。我们前期证实甲强龙通过下调内源性神经干细胞（eNSCs）中Notch受体1（NR1）表达，从而抑制eNSCs增殖，最终阻碍脊髓损伤后神经修复，但甲强龙作用eNSCs后导致细胞NR1表达下调的具体机制不清。预实验结果发现甲强龙可上调eNSCs中长链非编码RNA-NRA（LncRNA-NRA）表达，这是导致NR1表达下调、抑制eNSCs增殖的关键所在。我们通过基因芯片、RNA pull-down、RIP、生物学信息分析等手段明确LncRNA-NRA调控NR1表达的具体机制，及其在甲强龙抑制eNSCs增殖中的关键调控作用，为解决甲强龙在急性脊髓损伤中应用瓶颈提供新的理论依据。我们筛选出甲强龙影响细胞增殖过程中，起关键作用的LncRNA：9430037G07Rik，并筛选LncRNA起作用的的关键转录因子E2F1。并通过表达和敲低9430037G07Rik，验证其在甲强龙抑制细胞增殖中的重要作用，提供了新的lncRNA 9430037G07Rik和E2F1转录调控与甲强龙抑制神经干细胞增殖的证据。