Ye Guangyao, Zhang Yu, Zhao Jingsong, Chen Yuebo, Kong Lingsi, Sheng Chaoxu, Yuan Liyong
Department of Anesthesiology, Ningbo No. 6 Hospital, Ningbo, Zhejiang, PR China.
Neurol Res. 2020 Apr;42(4):299-307. doi: 10.1080/01616412.2020.1723313. Epub 2020 Feb 26.
: To explore the potential regulation mechanisms of miR-384-5p in Neuropathic pain (NP).: Rat model of chronic constriction injury (CCI) was established to induce NP in vivo. NP levels were assessed using Withdrawal Threshold (PWT) and Paw Withdrawal Latency (PWL). qPCR and Western blotting were used to determine the relative expression of miR-384-5p and SCN3A. The inflammation response in spinal microglia cells was determined by ELISA assay. Immunofluorescence assay was used to demonstrate the co-localization of miR-384-5p with SCN3A in rat dorsal root ganglions (DRGs). The target genes of miR-384-5p were verified by dual-luciferase report assays.: In the current study, the miR-384-5p expression level was significantly downregulated in CCI rats when comparing to the sham group. In addition, miR-384-5p agomir significantly repressed mechanical allodynia and heat hyperalgesia in CCI rats. Meanwhile, the current study indicated miR-384-5p could decrease inflammation progress in spinal microglia cells incubated in lipopolysaccharide. Consistently, overexpression of miR-384-5p obviously depressed inflammation cytokine levels in CCI rats. Dual-luciferase reporter assays indicated that SCN3A is a target gene of miR-384-5p.: miR-384-5p is a negative regulator in the development of neuropathic pain by regulating SCN3A, indicating that miR-384-5p might be a promising therapeutic target in the treatment of neuropathic pain. CCI: Chronic constriction injury; ZEB1: Zinc finger E box binding protein-1; MAPK6: Mitogen-activated protein kinase 6; COX-2: cyclooxygenase-2.
探索miR-384-5p在神经性疼痛（NP）中的潜在调控机制。建立慢性压迫损伤（CCI）大鼠模型以在体内诱导NP。使用撤针阈值（PWT）和缩爪潜伏期（PWL）评估NP水平。采用qPCR和蛋白质免疫印迹法测定miR-384-5p和SCN3A的相对表达。通过ELISA检测法测定脊髓小胶质细胞中的炎症反应。采用免疫荧光检测法证明miR-384-5p与SCN3A在大鼠背根神经节（DRG）中的共定位。通过双荧光素酶报告基因检测法验证miR-384-5p的靶基因。在本研究中，与假手术组相比，CCI大鼠中miR-384-5p表达水平显著下调。此外，miR-384-5p激动剂显著抑制了CCI大鼠的机械性异常性疼痛和热痛觉过敏。同时，本研究表明miR-384-5p可降低脂多糖孵育的脊髓小胶质细胞中的炎症进程。同样，miR-384-5p的过表达明显降低了CCI大鼠中的炎症细胞因子水平。双荧光素酶报告基因检测表明SCN3A是miR-384-5p的靶基因。miR-384-5p通过调节SCN3A在神经性疼痛发展中起负调节作用，表明miR-384-5p可能是治疗神经性疼痛的有前景的治疗靶点。CCI：慢性压迫损伤；ZEB1：锌指E盒结合蛋白-1；MAPK6：丝裂原活化蛋白激酶6；COX-2：环氧化酶-2。
