血清外泌体微小RNA-375-3p在吗啡依赖大鼠中的作用  

Effects of serum exosomal microRNA-375-3p in morphine-induced addiction rats

作　　者：李汉成 林泽鑫[1] 曾鹏 莫志贤 LI Han-cheng;LIN Ze-xin;ZENG Peng;MO Zhi-xian(School of Food and Pharmaceutical Engineering,Zhaoqing University,Zhaoqing 526061,Guangdong Province,China;School of Pharmacy,Zhaoqing Medical College,Zhaoqing 526020,Guangdong Province,China;School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,Guangdong Province,China)

机构地区：[1]肇庆学院食品与制药工程学院,广东肇庆526061 [2]肇庆医学高等专科学校药学院,广东肇庆526020 [3]南方医科大学中医药学院,广东广州510515

出　　处：《中国临床药理学杂志》2023年第11期1567-1571,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81873030、81673628);广东省重点领域研发计划基金资助项目(2021B0707010008);广东省科技厅农村科技特派员课题基金资助项目(KTP20200176);广东省教育厅青年创新人才基金资助项目(2017GkQNCX103);肇庆市科技计划基金资助项目(2022040314001);肇庆学院博士科研启动基金资助项目(220003);肇庆学院科研基金资助项目(QN202224);肇庆学院大学生创新创业训练计划基金资助项目(X202210580146)。

摘　　要：目的探讨血清外泌体微小RNA(miR)-375-3p在吗啡诱导大鼠成瘾中的可能作用机制。方法通过剂量递增法诱导吗啡依赖大鼠戒断模型。将20只大鼠随机分为正常组和模型组,模型组连续6 d皮下注射吗啡,剂量依次递增为5、10、20、40、60、80 mg·kg^(-1),正常组仅注射0.9%NaCl。用实时荧光聚合酶链反应(RT-PCR)检测血清外泌体miR-375-3p表达水平。通过Target Scan、miRDB和DIANA Tools预测miR-375-3p靶基因。用DAVID数据库对其靶基因进行基因本体(GO)及京都基因与基因组百科全书(KEGG)分析。通过STRING软件对其靶基因编码蛋白进行互作分析。通过Cytoscape软件构建miR-375-3p-靶基因-KEGG网络。结果正常组和模型组第1次戒断评分值分别为2.38±1.21和25.11±3.46,第2次分别为2.51±1.09和17.32±2.67;正常组和模型组纳洛酮第1次催促后体质量分别为(213.95±10.90)和(193.43±15.22)g,第2次分别为(217.09±14.49)和(181.23±14.76)g。模型组与正常组比较,2次戒断评分值及体质量差异均有统计学意义(P<0.01,P<0.05)。正常组和模型组血清外泌体miR-375-3p相对表达水平分别为5.70±2.33和13.56±5.18。对miR-375-3p的306个潜在靶基因集合分析发现,其分子功能主要富集于神经元的产生、中枢神经系统发育等生物学过程(P<0.05),涉及谷氨酸能突触及多巴胺能突触等多条信号转导通路(P<0.05)。miR-375-3p靶基因编码蛋白之间存在复杂的相互作用,靶基因L型电压依赖钙离子通道α1C(CACNA1C)、鸟嘌呤核苷酸结合蛋白γ11(GNG11)、谷氨酸受体红藻氨酸离子5(GRIK5)、盘状大同源物3(DLG3)等编码蛋白在互作网络中具有核心地位。结论miR-375-3p可能通过调控中枢神经系统发育等生物过程作用于谷氨酸能突触及多巴胺能突触等信号通路进而调节下游靶蛋白参与吗啡成瘾的发生过程。Objective To investigate the possible mechanism of serum exosomal microRNA(miR)-375-3p in morphine-induced addiction rats.Methods A morphine-dependent rat withdrawal model was induced by dose-escalation method.Twenty rats were randomly divided into normal and model groups,and model group was injected with morphine subcutaneously for 6 consecutive days at increasing doses of 5,10,20,40,60 and 80 mg·kg^(-1),while normal group was only injected with 0.9%NaCl.Serum exosomal miR-375-3p expression was detected by reverse transcription-polymerase chain reaction(RT-PCR).The miR-375-3p target gene was predicted by Target Scan,miRDB and DIANA Tools.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)analysis of its target genes were performed using DAVID database.Interaction analysis of the proteins encoded by its target genes was performed by STRING software.The miR-375-3p-target gene-KEGG network was constructed by Cytoscape software.Results The first withdrawal score values of normal group and model group were 2.38±1.21 and 25.11±3.46,respectively,and 2.51±1.09 and 17.32±2.67 for the second time,respectively;the body weight after the first naloxone rush was(213.95±10.90)and(193.43±15.22)g,respectively,and the second(217.09±14.49)and(181.23±14.76)g,respectively.Compared with normal group,the two withdrawal score values and weight changes were statistically different in model group(P<0.01,P<0.05).Serum exosomal miR-375-3p expression of normal group and model group was 5.70±2.33 and 13.56±5.18,respectively.Bioinformatics analysis of the set of 306 potential target genes of miR-375-3p revealed that its molecular functions were mainly enriched in biological processes such as neuronal production and CNS development(P<0.05),and involved multiple signaling pathways such as glutamatergic synapses and dopaminergic synapses(P<0.05).miR-375-3p target genes encode proteins with complex interactions,and the target genes calcium channel voltage-dependent L-type subunit alpha-1C(CACNA1C),guanine nucleotide-bind

关 键 词：吗啡 药物依赖 微小RNA-375-3p 血清外泌体 生物功能分析 

分 类 号：R971[医药卫生—药品]