BACKGROUND Neuropathic pain (NP) is the comorbidity in spinal cord injury(SCI), which is the hardest to cure. Non-coding RNA dysregulations are related to the development of NP. NEAT1(nuclear paraspeckle assembly transcript 1) is a new type of lncRNA. This study explores the role and specific mechanism of NEAT1 in SCI-mediated NP. METHODS Firstly, the NEAT1 expression in SCI rats and the control group was detected with RT-PCR to analyze the relationship between NEAT13 and NP symptoms. Then, SCI rats were intrathecally injected with NEAT13 overexpressing and knocking down lentiviruses. Afterward, ELISA was utilized to assess the expression of IL-6, IL-1β and TNFα in rats. Subsequently, immunohistochemistry was adopted to verify the activation of microglial cells. After that, bioinformatics analysis was employed to further predict the downstream target genes of NEAT1, while RT-PCR and Western blot were conducted to determine the relative expression of miR-128-3p and aquaporin-4(AQP4). Meanwhile, a dual-luciferase reporter assay was performed to further study the targeting relationship between NEAT1 and miR-128-3p, and miR-128-3p and AQP4. RESULTS SCI rats showed distinctly higher NEAT1 expression compared with that of the control group. ELISA experiment confirmed that the over-expression of NEAT1 enhanced the expression of IL-6, IL-1β, and TNFα in SCI rats. Other related mechanism studies revealed that NEAT13 targeted and inhibited miR-128-3p as its competing endogenous RNA (ceRNA), and enhanced AQP4 expression, while miR-128-3p targeted AQP4 to regulate its expression. SUMMARY NEAT1 affects AQP4 signaling pathway to alleviate the spinal cord injury-induced NP via promoting miR-128-3p expression.

中文翻译：

---

LncRNA NEAT1/miR-128-3p/AQP4轴调节脊髓损伤诱导的神经病理性疼痛进展

背景神经病理性疼痛（NP）是脊髓损伤（SCI）中最难治愈的并发症。非编码 RNA 失调与 NP 的发生有关。NEAT1（nuclear paraspeckle组装转录本1）是一种新型的lncRNA。本研究探讨了 NEAT1 在 SCI 介导的 NP 中的作用和具体机制。方法首先采用RT-PCR检测SCI大鼠和对照组NEAT1的表达，分析NEAT13与NP症状的关系。然后，SCI 大鼠鞘内注射 NEAT13 过表达和击倒慢病毒。之后，ELISA被用来评估IL-6、IL-1β和TNFα在大鼠中的表达。随后，采用免疫组织化学来验证小胶质细胞的活化。之后，采用生物信息学分析进一步预测NEAT1下游靶基因，RT-PCR和Western blot检测miR-128-3p和水通道蛋白4(AQP4)的相对表达。同时，通过双荧光素酶报告基因检测进一步研究了NEAT1与miR-128-3p、miR-128-3p与AQP4之间的靶向关系。结果 与对照组相比，SCI 大鼠表现出明显更高的 NEAT1 表达。ELISA实验证实NEAT1的过表达增强了SCI大鼠IL-6、IL-1β和TNFα的表达。其他相关机制研究表明，NEAT13靶向并抑制miR-128-3p作为其竞争性内源RNA（ceRNA），增强AQP4表达，而miR-128-3p靶向AQP4以调节其表达。