下调长链非编码 RNA DLEU1 通过靶向 miR-133a-3p/SRPK1 轴减轻大鼠慢性缩窄性损伤诱导的神经病理性疼痛敏感性。
Downregulation of long noncoding RNA DLEU1 attenuates hypersensitivity in chronic constriction injury-induced neuropathic pain in rats by targeting miR-133a-3p/SRPK1 axis.
机构信息
Department of Anesthesiology, Hunan Provincial Maternal and Child Health Care Hospital, Changsha, 410008, Hunan, China.
Department of Burn and Plastic Surgery, Xiangya Hospital Central South University, No. 87 Xiangya Road, Kaifu District, Changsha, 410008, Hunan, China.
出版信息
Mol Med. 2020 Nov 10;26(1):104. doi: 10.1186/s10020-020-00235-6.
BACKGROUND
Neuropathic pain belongs to chronic pain and is caused by the primary dysfunction of the somatosensory nervous system. Long noncoding RNAs (lncRNAs) have been reported to regulate neuronal functions and play significant roles in neuropathic pain. DLEU1 has been indicated to have close relationship with neuropathic pain. Therefore, our study focused on the significant role of DLEU1 in neuropathic pain rat models.
METHODS
We first constructed a chronic constrictive injury (CCI) rat model. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were employed to evaluate hypersensitivity in neuropathic pain. RT-qPCR was performed to analyze the expression of target genes. Enzyme-linked immunosorbent assay (ELISA) was conducted to detect the concentrations of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-1β. The underlying mechanisms of DLEU1 were investigated using western blot and luciferase reporter assays.
RESULTS
Our findings showed that DLEU1 was upregulated in CCI rats. DLEU1 knockdown reduced the concentrations of IL-6, IL-1β and TNF-α in CCI rats, suggesting that neuroinflammation was inhibited by DLEU1 knockdown. Besides, knockdown of DLEU1 inhibited neuropathic pain behaviors. Moreover, it was confirmed that DLEU1 bound with miR-133a-3p and negatively regulated its expression. SRPK1 was the downstream target of miR-133a-3p. DLEU1 competitively bound with miR-133a-3p to upregulate SRPK1. Finally, rescue assays revealed that SRPK1 overexpression rescued the suppressive effects of silenced DLEU1 on hypersensitivity in neuropathic pain and inflammation of spinal cord in CCI rats.
CONCLUSION
DLEU1 regulated inflammation of the spinal cord and mediated hypersensitivity in neuropathic pain in CCI rats by binding with miR-133a-3p to upregulate SRPK1 expression.
背景
神经病理性疼痛属于慢性疼痛，由躯体感觉神经系统的原发功能障碍引起。长链非编码 RNA（lncRNA）已被报道可调节神经元功能，并在神经病理性疼痛中发挥重要作用。DLEU1 已被证明与神经病理性疼痛密切相关。因此，我们的研究重点是 DLEU1 在神经病理性疼痛大鼠模型中的重要作用。
方法
我们首先构建了慢性压迫性损伤（CCI）大鼠模型。利用足底缩足反射阈值（PWT）和足底缩足潜伏期（PWL）评估神经病理性疼痛的敏感性。采用 RT-qPCR 分析靶基因的表达。酶联免疫吸附试验（ELISA）检测白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）和白细胞介素 1β（IL-1β）的浓度。通过 Western blot 和荧光素酶报告基因实验研究 DLEU1 的潜在机制。
结果
我们的研究结果表明，DLEU1 在 CCI 大鼠中上调。DLEU1 敲低降低了 CCI 大鼠中 IL-6、IL-1β 和 TNF-α的浓度，表明 DLEU1 敲低抑制了神经炎症。此外，DLEU1 敲低抑制了神经病理性疼痛行为。此外，证实 DLEU1 与 miR-133a-3p 结合并负调控其表达。SRPK1 是 miR-133a-3p 的下游靶标。DLEU1 与 miR-133a-3p 竞争性结合，上调 SRPK1。最后，挽救实验表明，SRPK1 过表达挽救了沉默 DLEU1 对 CCI 大鼠神经病理性疼痛和脊髓炎症敏感性的抑制作用。
结论
DLEU1 通过与 miR-133a-3p 结合上调 SRPK1 表达，调节脊髓炎症，介导 CCI 大鼠神经病理性疼痛的敏感性。
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