MicroRNA-513b-5p调控mGluR5表达对紫杉醇诱导神经病理性疼痛大鼠的影响  

Effect of MicroRNA-513b-5p on Paclitaxel-induced Neuropathic Pain in Rats by Regulating the Expression of mGluR5

作　　者：彭宇川 张世豪 柯均一 张硕雅 肖昀 PENG Yu-chuan;ZHANG Shi-hao;KE Jun-yi;ZHANG Shuo-ya;XIAO Yun(Department of Anesthesiology,Renmin Hospital,Hubei University of Medicine,Shiyan,Hubei 442000;Department of Anesthesiology,theFourth Affiliated Hospital,School of Medicine,Zhejiang University,Yiwu,Zhejiang 322000,China)

机构地区：[1]湖北医药学院附属人民医院麻醉手术中心,湖北十堰442000 [2]浙江大学医学院附属第四医院麻醉科,浙江义乌322000

出　　处：《湖北医药学院学报》2022年第6期549-554,560,共7页Journal of Hubei University of Medicine

基　　金：湖北省自然科学基金青年项目(ZRMS2018001630);湖北省卫生健康委员会项目(WJ2021Q009);十堰市科技计划项目(21Y69);湖北医药学院研究生科技创新项目(YC2021042)。

摘　　要：目的:研究MicroRNA(miR)-513b-5p对代谢型谷氨酸受体5(mGluR5)的调节关系及其对紫杉醇诱导神经病理性疼痛大鼠的影响。方法:RT-qPCR和Western blot法检测紫杉醇诱导神经病理性疼痛大鼠DRG中miR-513b-5p和mGluR5的表达变化。根据生物信息学分析、双荧光素酶报告基因实验和RNA pull-down实验验证miR-513b-5p和mGluR5相互作用。大鼠紫杉醇造模同时行髓鞘内置管,然后按不同分组进行慢病毒髓鞘内注射:⑴对照组(Lv-NC组);⑵miRNA非特异性对照组(Lv-miRNA-NC组);⑶过表达miR513b-5p组(Lv-miR513b-5p组);⑷过表达mGluR5组(Lv-mGluR5组);⑸过表达miR513b-5p混合mGluR5组(Lv-mGluR5+Lv-miR513b-5p组),1周后使用Western blot法检测各组DRG中mGluR5水平,并后续进行热痛觉阈值检测及机械痛觉敏感性检测。结果:miR-513b-5p在紫杉醇疼痛模型大鼠DRG中表达水平明显降低,而mGluR5的水平明显升高(P<0.05),并且mGluR5的蛋白水平在造模后明显上调(P<0.05)。StarBase在线分析数据库(http://starbase.sysu.edu.cn/)显示mGluR5与miR-513b-5p存在互补位点,双荧光素酶报告基因实验和RNA pull-down实验均证实miR-513b-5p能特异结合mGluR5(P<0.05);鞘内注射慢病毒Lv-miR513b-5p在降低mGluR5蛋白表达水平同时,行为学结果显示大鼠TWL及MWT明显升高(P<0.05)。结论:miR-513b-5p可以通过靶向负调控大鼠DRG中mGluR5的表达水平,从而缓解紫杉醇诱导的神经病理性疼痛。Objective To investigate the regulatory relationship of microRNA(miR)-513b-5p on metabotropic glutamate receptor 5(mGluR5)and its effect on paclitaxel-induced neuropathic pain in rats.Methods RT-qPCR and Western Blot were performed to detect the expression changes of miR-513b-5p and mGluR5 in DRG of paclitaxel-induced neuropathic pain rats.Based on bioinformatics analysis,dual luciferase reporter gene assay and RNA pull-down assay were performed to verify the miR-513b-5p and mGluR5 interaction.Rats were modeled with paclitaxel and underwent intramyelin intubation,followed by intramyelin injection of lentivirus according to different groups:⑴control group(Lv-NC group),⑵miRNA non-specific control group(Lv-miRNA-NC group),⑶Overexpression of miR513b-5p group(Lv-miR513b-5p group),⑷Overexpression of mGluR5 group(Lv-mGluR5 group),⑸Overexpression of miR513b-5p mixed mGluR5 group(Lv-mGluR5+Lv-miR513b-5p group).After one week,Western blot was used to detect the level of mGluR5 in DRG of each group,followed by thermal pain threshold detection and mechanical pain sensitivity detection.Results The expression level of miR-513b-5p in the DRG of paclitaxel-induced pain model rats was significantly decreased,while the level of mGluR5 was significantly increased(P<0.05),and the protein level of mGluR5 was significantly increased after modeling(P<0.05).StarBase online analysis database(http://starbase.sysu.edu.cn/)showed that there were complementary sites between mGluR5 and miR-513b-5p.Both dual luciferase reporter gene experiment and RNA pull-down experiment confirmed that miR-513b-5p could specifically bind mGluR5(P<0.05).Intrathecal injection of lentivirus Lv miR513b-5p decreased the expression level of mGluR5 protein,and behavioral results showed that TWL and MWT in rats were significantly increased(P<0.05).Conclusion miR-513b-5p can alleviate paclitaxel-induced neuropathic pain by negatively regulating the expression of mGluR5 in rat DRG.

关 键 词：MicroRNA-513b-5p 代谢型谷氨酸受体5 紫杉醇 化疗所致周围神经病变 神经病理性疼痛 

分 类 号：R285.5[医药卫生—中药学]