Wang Xiaopeng, Pang Jun, Cui Jian, Liu Aifen, Wang Hui
Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University, Taiyuan 030032, China.
Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University, Taiyuan 030032, China.
Transpl Immunol. 2023 Feb;76:101735. doi: 10.1016/j.trim.2022.101735. Epub 2022 Nov 2.
BACKGROUND/PURPOSE: Neuropathic pain(NP) is derived from the dysfunctions of nerve system. The current research is to explore the impact and mechanism of miR-19a-3p in neuropathic pain in rats.
The NP was induced through the chronic constriction injury (CCI) surgery in rats. The pro-inflammatory factors (IL-1β, IL-6, TNF-α) in spinal cord tissues from rats were measured using Elisa kits. Moreover, the different levels of thermal hyperalgesia and mechanical allodynia in rats were examined through paw withdrawal latency (PWL) and paw withdrawal threshold (PWT). To investigate into the role of miR-19a-3p and KLF7 in NP of rats, the knockdown of miR-19a-3p alone or along with KLF7 downregulation in rats were achieved through lentivirus injection. The miR-19a-3p and KLF7 expression in spinal cord of rats on Day 3,7,14 after CCI were detected using RT-qPCR. The protein expression of KLF7 were measured by Western blot. Bioinformatics and luciferase assays were used for the prediction and verification of bindings between KLF7 and miR-19a-3p.
CCI surgery caused neuropathic pain in rats with the levels of inflammatory cytokines increased and PWL and PWT decreased. Moreover, miR-19a-3p expression was increased while the protein and mRNA levels were decreased in spinal cord tissues in rats after CCI surgery. In rat microglial cells, miR-19a-3p downregulation could promote the KLF7 in both mRNA and protein expression. In spinal cord tissues of rats, the inhibition of miR-19a-3p enhanced the KLF7 expression. Furthermore, miR-19a-3p downregulation suppressed the IL-1β, IL-6 and TNF-α concentrations, and could decrease the NP but inhibition of KLF7 could partially reverse this in CCI rats.
miR-19a-3p inhibition may alleviate NP via KLF7 in CCI rats.
背景/目的：神经性疼痛（NP）源于神经系统功能障碍。当前研究旨在探讨miR-19a-3p在大鼠神经性疼痛中的作用及其机制。
通过慢性缩窄损伤（CCI）手术诱导大鼠产生神经性疼痛。使用酶联免疫吸附测定（ELISA）试剂盒检测大鼠脊髓组织中的促炎因子（IL-1β、IL-6、TNF-α）。此外，通过爪部撤离潜伏期（PWL）和爪部撤离阈值（PWT）检测大鼠不同程度的热痛觉过敏和机械性异常性疼痛。为研究miR-19a-3p和KLF7在大鼠神经性疼痛中的作用，通过慢病毒注射分别单独敲低大鼠体内的miR-19a-3p以及同时敲低miR-19a-3p和KLF7。采用逆转录-定量聚合酶链反应（RT-qPCR）检测CCI术后第3、7、14天大鼠脊髓中miR-19a-3p和KLF7的表达。通过蛋白质免疫印迹法检测KLF7的蛋白表达。利用生物信息学和荧光素酶报告基因检测对KLF7与miR-19a-3p之间的结合进行预测和验证。
CCI手术导致大鼠出现神经性疼痛，炎症细胞因子水平升高，PWL和PWT降低。此外，CCI术后大鼠脊髓组织中miR-19a-3p表达增加，而KLF7的蛋白和mRNA水平降低。在大鼠小胶质细胞中，下调miR-19a-3p可促进KLF7的mRNA和蛋白表达。在大鼠脊髓组织中，抑制miR-19a-3p可增强KLF7的表达。此外，下调miR-19a-3p可抑制IL-1β、IL-6和TNF-α的浓度，并减轻神经性疼痛，但在CCI大鼠中抑制KLF7可部分逆转这种作用。
在CCI大鼠中，抑制miR-19a-3p可能通过KLF7减轻神经性疼痛。
