Department of Pain Medicine, Shenzhen People's Hospital, The First Affiliated Hospital of Southern University of Science and Technology, Shenzhen, China.
Department of Anesthesiology, The Affiliated Wuxi No.2 People's Hospital of Nanjing Medical University, Wuxi, China.
Ann Palliat Med. 2021 Nov;10(11):11566-11577. doi: 10.21037/apm-21-2651.
This study investigated the role and molecular mechanisms of the long intergenic non-protein coding RNA 472 (LINC00472) in neuropathic pain using a chronic constrictive injury (CCI) rat model.
CCI rat model was established and PC12 cells were induced by LPS to simulate neuropathological injury in vivo and in vitro. The levels of LINC00472, miR-300, and high mobility group box protein 1 (HMGB1) in the spinal cord tissue of CCI rats and PC12 pheochromocytoma cells were assessed by qRT-PCR and western blot. The effects of LINC00472 on neuropathic pain in the CCI rats were observed by their pain behavior. ELISA was used to detect the levels of inflammatory cytokines in rat tissues and cells. The molecular mechanisms of LINC00472 were verified by luciferase experiments, RNA immunoprecipitation, and RNA pull down assays.
The expression of LINC00472 and HMGB1 were upregulated, and the expression of miR-300 was downregulated in the spinal cord tissues of CCI rats and in PC12 cells. The upregulation of LINC00472 in CCI rats significantly induced the occurrence of neuropathic pain. In addition, downregulation of LINC00472 inhibited the inflammatory response of CCI rats and PC12 cells. This study identified miR-300 as a target gene of LINC00472, and HMGB1 as the target gene of miR-300. Further experiments confirmed that the expression of anti-miR-300 could partially reverse the anti-inflammatory effects and the reduction of neuropathic pain induced by low expression of LINC00472.
LINC00472 promotes the progression of neuropathic pain by reducing miR-300 expression and increasing HMGB1 expression. The LINC00472/miR-300/HMGB1 axis may be a novel therapeutic target for neuropathic pain.
本研究通过慢性压迫性损伤（CCI）大鼠模型，探讨了长链非编码 RNA 472（LINC00472）在神经病理性疼痛中的作用和分子机制。
建立 CCI 大鼠模型，用 LPS 诱导 PC12 细胞，在体内和体外模拟神经病理损伤。采用 qRT-PCR 和 Western blot 检测 CCI 大鼠脊髓组织和 PC12 嗜铬细胞瘤细胞中 LINC00472、miR-300 和高迁移率族蛋白 1（HMGB1）的水平。通过 CCI 大鼠的疼痛行为观察 LINC00472 对神经病理性疼痛的影响。ELISA 检测大鼠组织和细胞中炎症细胞因子的水平。通过荧光素酶实验、RNA 免疫沉淀和 RNA 下拉实验验证 LINC00472 的分子机制。
CCI 大鼠脊髓组织和 PC12 细胞中 LINC00472 和 HMGB1 的表达上调，miR-300 的表达下调。CCI 大鼠 LINC00472 的上调显著诱导了神经病理性疼痛的发生。此外，下调 LINC00472 抑制了 CCI 大鼠和 PC12 细胞的炎症反应。本研究确定 miR-300 是 LINC00472 的靶基因，HMGB1 是 miR-300 的靶基因。进一步的实验证实，抗 miR-300 的表达可以部分逆转低表达 LINC00472 诱导的抗炎作用和神经病理性疼痛的减轻。
LINC00472 通过降低 miR-300 的表达和增加 HMGB1 的表达促进神经病理性疼痛的进展。LINC00472/miR-300/HMGB1 轴可能是治疗神经病理性疼痛的新靶点。
