MiR-21-5p通过下调电压门控钾离子通道Kv1.1表达减轻大鼠三叉神经痛  

MiR-21-5p alleviates trigeminal neuralgia in rats through down-regulation of voltage-gated potassium channel Kv1.1

作　　者：周雪雯 郭刚文[1] 余珊子 胡蓉[1] ZHOU Xuewen;GUO Gangwen;YU Shanzi;HU Rong(Department of Pain,Third Xiangya Hospital,Central South University,Changsha 410013;Department of Anesthesiology,First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310009;Department of Anesthesiology,Hunan Provincial People’s Hospital(First Hospital Affiliated with Hunan Normal University),Changsha 410002,China)

机构地区：[1]中南大学湘雅三医院疼痛科,长沙410013 [2]浙江大学医学院附属第一医院麻醉科,杭州310009 [3]湖南省人民医院(湖南师范大学附属第一医院)麻醉科,长沙410002

出　　处：《中南大学学报（医学版）》2024年第1期29-39,共11页Journal of Central South University ：Medical Science

基　　金：湖南省自然科学基金(2019JJ40457)。

摘　　要：目的:三叉神经痛(trigeminal neuralgia,TN)是一种临床上常见的神经病理性疼痛。电压门控性钾通道(voltage-gated potassium channel,Kv)已被证实参与TN的发生、发展,但具体机制仍不明确。微RNA(microRNA,miR)可通过调节三叉神经节(trigeminal ganglion,TG)上Kv通道的表达及神经元兴奋性,参与神经病理性疼痛。本研究旨在探索TN模型中TG上Kv1.1和miR-21-5p的关系,评估miR-21-5p是否对Kv1.1有调控作用,为TN的治疗提供新的靶点。方法:将48只SD大鼠随机分为6组:1)假手术组(sham组,n=12),大鼠仅在术侧切口缝合,不结扎神经;2)Sham+agomir NC组(n=6),sham大鼠通过脑立体定位注射方法于术侧TG微量注射agomir NC;3)Sham+miR-21-5p agomir组(n=6),sham大鼠通过脑立体定位注射方法于术侧TG微量注射miR-21-5p agomir;4)TN组(n=12),采用铬肠线慢性缩窄性眶下远端神经损伤(chronic constriction injury of the distal infraorbital nerve,dIoN-CCI)法构建TN大鼠模型;5)TN+antagomir NC组(n=6),TN大鼠通过脑立体定位注射方法于术侧TG微量注射antagomir NC;6)TN+miR-21-5p antagomir组(n=6),TN大鼠通过脑立体定位注射方法于术侧TG微量注射miR-21-5p antagomir。检测术后各组大鼠面部机械痛阈变化。采用蛋白质印迹法和实时反转录聚合酶链反应检测术后大鼠术侧TG中Kv1.1和miR-21-5p的表达情况。利用双荧光素酶报告基因确定Kv1.1和miR-21-5p是否存在靶标关系,即miR-21-5p是否可以直接影响KCNA1的3'端非翻译区(3'-untranslated region,3'-UTR)。通过免疫荧光法测定,对脑立体定位注射的效果进行评价,随后分别将miR-21-5p的类似物(agomir)和agomir NC通过脑立体定位仪注射至sham组大鼠TG内,使miR-21-5p过表达;向TN组大鼠TG内分别注射miR-21-5p的抑制剂(antagomir)和antagomir NC,抑制miR-21-5p表达。观察给药前后大鼠行为学变化,检测干预后大鼠TG内miR-21-5p和Kv1.1表达的变化。结果:与基础痛阈值相比,TN组大鼠在术后第Objective:Trigeminal neuralgia(TN)is a common neuropathic pain.Voltage-gated potassium channel(Kv)has been confirmed to be involved in the occurrence and development of TN,but the specific mechanism is still unclear.MicroRNA may be involved in neuropathic pain by regulating the expression of Kv channels and neuronal excitability in trigeminal ganglion(TG).This study aims to explore the relationship between Kv1.1 and miR-21-5p in TG with a TN model,evaluate whether miR-21-5p has a regulatory effect on Kv1.1,and to provide a new target and experimental basis for the treatment of TN.Methods:A total of 48 SD rats were randomly divided into 6 groups:1)a sham group(n=12),the rats were only sutured at the surgical incision without nerve ligation;2)a sham+agomir NC group(n=6),the sham rats were microinjected with agomir NC through stereotactic brain injection in the surgical side of TG;3)a sham+miR-21-5p agomir group(n=6),the sham rats were microinjected with miR-21-5p agomir via stereotactic brain injection in the surgical side of TG;4)a TN group(n=12),a TN rat model was constructed using the chronic constriction injury of the distal infraorbital nerve(dIoN-CCI)method with chromium intestinal thread;5)a TN+antagonist NC group(n=6),TN rats were microinjected with antagonist NC through stereotactic brain injection method in the surgical side of TG;6)a TN+miR-21-5p antagonist group(n=6),TN rats were microinjected with miR-21-5p antagonist through stereotactic brain injection in the surgical side of TG.The change of mechanical pain threshold in rats of each group after surgery was detected.The expressions of Kv1.1 and miR-21-5p in the operative TG of rats were detected by Western blotting and real-time reverse transcription polymerase chain reaction.Dual luciferase reporter genes were used to determine whether there was a target relationship between Kv1.1 and miR-21-5p and whether miR-21-5p directly affected the 3'-UTR terminal of KCNA1.The effect of brain stereotaxic injection was evaluated by immunofluorescence assay,and

关 键 词：电压门控性钾通道 miR-21-5p 三叉神经痛 三叉神经节 

分 类 号：R745.11[医药卫生—神经病学与精神病学]