Department of Pain Medicine, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Department of Anesthesiology, Guangdong Women and Children Hospital, Guangzhou, China.
Neurol Res. 2024 Jan;46(1):54-64. doi: 10.1080/01616412.2023.2257454. Epub 2023 Dec 8.
Diabetic neuropathic pain (DNP) is a serious complication for diabetic patients involving nervous system. MicroRNAs (miRNAs) are small-noncoding RNAs which are dysregulated in neuropathic pain, and might be critical molecules for pain treatment. Our previous study has shown miR-184-5p was significantly downregulated in DNP. Therefore, the mechanism of miR-184-5p in DNP was investigated in this study.
A DNP model was established through streptozotocin (STZ). The pharmacological tools were injected intrathecally, and pain behavior was evaluated by paw withdrawal mechanical thresholds (PWMTs). Bioinformatics analysis, Dual-luciferase reporter assay and fluorescence-in-situ-hybridization (FISH) were used to seek and confirm the potential target genes of miR-184-5p. The expression of relative genes and proteins was analyzed by quantitative reverse transcriptase real-time PCR (qPCR) and western blotting.
MiR-184-5p expression was down-regulated in spinal dorsal on days 7 and 14 after STZ, while intrathecal administration of miR-184-5p agomir attenuates neuropathic pain induced by DNP and intrathecal miR-184-5p antagomir induces pain behaviors in naïve mice. Chemokine CC motif ligand 1 (CCL1) was found to be a potential target of miR-184-5p and the protein expression of CCL1 and the mRNA expression of CCR8 were up-regulated in spinal dorsal on days 7 and 14 after STZ. The luciferase reporter assay and FISH demonstrated that CCL1 is a direct target of miR-184-5p. MiR-184-5p overexpression attenuated the expression of CCL1/CCR8 in DNP; intrathecal miR-184-5p antagomir increased the expression of CCL1/CCR8 in spinal dorsal of naïve mice.
This research illustrates that miR-184-5p alleviates DNP through the inhibition of CCL1/CCR8 signaling expression.
糖尿病性神经病理性疼痛（DNP）是糖尿病患者的一种严重并发症，涉及神经系统。microRNAs（miRNAs）是一种表达失调的小非编码 RNA，与神经病理性疼痛有关，可能是疼痛治疗的关键分子。我们之前的研究表明，miR-184-5p 在 DNP 中显著下调。因此，本研究旨在探讨 miR-184-5p 在 DNP 中的作用机制。
通过链脲佐菌素（STZ）建立 DNP 模型。通过鞘内注射药理学工具，通过爪撤回机械阈值（PWMT）评估疼痛行为。生物信息学分析、双荧光素酶报告基因检测和荧光原位杂交（FISH）用于寻找和确认 miR-184-5p 的潜在靶基因。通过定量逆转录实时 PCR（qPCR）和 Western blot 分析相对基因和蛋白的表达。
miR-184-5p 在 STZ 后第 7 天和第 14 天在脊髓背角表达下调，而鞘内给予 miR-184-5p 激动剂可减轻 DNP 引起的神经病理性疼痛，鞘内给予 miR-184-5p 拮抗剂可引起正常小鼠的疼痛行为。趋化因子 CC 基序配体 1（CCL1）被发现是 miR-184-5p 的潜在靶标，并且在 STZ 后第 7 天和第 14 天脊髓背角中 CCL1 蛋白表达和 CCR8mRNA 表达上调。荧光素酶报告基因检测和 FISH 表明 CCL1 是 miR-184-5p 的直接靶标。miR-184-5p 过表达可减轻 DNP 中 CCL1/CCR8 的表达；鞘内给予 miR-184-5p 拮抗剂可增加正常小鼠脊髓背角中 CCL1/CCR8 的表达。
本研究表明，miR-184-5p 通过抑制 CCL1/CCR8 信号表达来缓解 DNP。
