miR-218-5p改善帕金森病小鼠多巴胺能神经元变性  

MiR-218-5p Ameliorates Degeneration of Dopaminergic Neurons in MPTP-Induced Mice

作　　者：王丹蕾 薛峥 覃奇雄 李静怡 赵经纬 毛志娟 WANG Dan-lei;XUE Zheng;QIN Qi-xiong;LI Jing-yi;ZHAO Jing-wei;MAO Zhi-juan(Department of Neurology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《神经损伤与功能重建》2022年第5期249-253,共5页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金重大研究项目(No.91849121);国家自然科学基金青年科学基金(No.81901303)。

摘　　要：目的:探索miR-218-5p对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的急性帕金森病(PD)模型小鼠黑质多巴胺能神经元的保护作用及其可能机制。方法:18只8周雄性C57小鼠随机分配至MPTP组和PBS组,分别腹腔注射20 mg/kg MPTP或等体积无菌PBS,每隔2 h注射一次,共4次。另将36只8周雄性C57小鼠随机平分为(NC agomir+PBS)组(NCPBS组)、(NC agomir+MPTP)组(NCMPTP组)、(miR-218-5p agomir+PBS)组(218PBS组)、(miR-218-5p agomir+MPTP)组(218MPTP组),每组9只,分别进行双侧黑质立体定位注射miR-218-5p agomir/NC agomir,3 d后腹腔注射MPTP或PBS。免疫荧光染色和Western blot检测小鼠黑质多巴胺能神经元酪氨酸羟化酶(TH)的表达;荧光定量PCR检测小鼠黑质miR-218-5p和Wnt7a、Ctnnb1、Lef1、Birc5 mRNA的表达。结果:与PBS组相比,MPTP组小鼠黑质miR-218-5p表达显著下降(P<0.05);与NCPBS组相比,NCMPTP组TH+细胞数量和TH蛋白表达减少(P<0.05),Wnt7a、Ctnnb1、Lef1、Birc5 mRNA表达下降(P<0.05);与NCMPTP组相比,218MPTP组TH+细胞数量和TH蛋白表达增加(P<0.05),Wnt7a、Ctnnb1、Lef1、Birc5 mRNA表达上升(P<0.05)。结论:miR-218-5p可能通过调控Wnt/β-catenin信号通路逆转MPTP诱导的急性PD模型小鼠黑质多巴胺能神经元变性,提示miR-218-5p有可能成为PD的潜在治疗靶点。Objective:To investigate the protective effect of miR-218-5p on dopaminergic neurons in the substantia nigra compacta(SNc)by establishing a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced mouse model of Parkinson’s disease(PD).Methods:A total of 188-week-old male C57 mice were randomly divided into MPTP group and PBS group.Mice were injected intraperitoneally with MPTP(20 mg/kg)or with an equal amount of PBS every 2 h for a total of 4 doses.An additional 368-week-old adult male C57 mice were randomly divided into 4 groups with 9 mice in each:(NC agomir+PBS)(NCPBS)group,(NC agomir+MPTP)(NCMPTP)group,(miR-218-5p agomir+PBS)(218PBS)group,and(miR-218-5p agomir+MPTP)(218MPTP)group.Bilateral stereotaxic injections of miR-218-5p agomir or NC agomir in the SNc were performed respectively,then 3 days later,mice were injected intraperitoneally with MPTP or PBS as before.The expression of tyrosine hydroxylase(TH)protein in the SNc was detected by immunofluorescence and Western blot(WB).The expression of miR-218-5p,Wnt7a mRNA,Ctnnb1 mRNA,Lef1 mRNA,and Birc5 mRNA in the SNc was assessed with quantitative real-time PCR(qPCR).Results:Compared with the PBS group,the expression of miR-218-5p of SNc in the MPTP group decreased obviously(P<0.05).Compared with NCPBS group mice,the NCMPTP group mice showed a reduced number of TH+cells,decreased expression of TH protein,and decreased expression of Wnt7a,Ctnnb1,Lef1,and Birc5 mRNA in the SNc(P<0.05).Compared with NCMPTP group mice,the 218MPTP group mice had a greater number of TH+cells,increased expression of TH protein,and increased expression of Wnt7a,Ctnnb1,Lef1,and Birc5 mRNA(P<0.05).Conclusion:MiR-218-5p significantly alleviates MPTP-induced dopaminergic neuronal degeneration in the SNc,potentially via Wnt/β-catenin signaling.This finding may provide a potential therapeutic target for the treatment of PD.

关 键 词：帕金森病 多巴胺能神经元 miR-218-5p Wnt/β-catenin信号 

分 类 号：R741[医药卫生—神经病学与精神病学] R742.5[医药卫生—临床医学]