钩藤碱对甲基苯丙胺依赖大鼠位置偏爱和血清外泌体miRNAs表达的影响  

Effects of rhynchophylline on conditioned place preference and expression of serum exosome miRNAs in methamphetamine-dependent rats

作　　者：李汉成 林泽鑫[1] 蒋昭 莫志贤 LI Han-cheng;LIN Ze-xin;JIANG Zhao;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8期1137-1141,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：目的观察钩藤碱对甲基苯丙胺依赖大鼠位置偏爱效应及其血清外泌体中miRNAs表达的影响。方法通过甲基苯丙胺诱导大鼠条件性位置偏爱(CPP)模型。将30只大鼠随机分为正常组、模型组和实验组。模型组和实验组用2 mg·kg^(-1)甲基苯丙胺皮下注射进行造模,正常组仅注射0.9%NaCl,同时实验组用60 mg·kg^(-1)钩藤碱腹腔注射进行干预治疗。用蛋白质印迹法检测其生物标志物的表达情况,用基因芯片技术检测血清外泌体miRNAs表达谱。通过sRNA bench软件计算并校正miRNAs的表达量,用sugar-edge study软件分析各组间存在差异表达的miRNAs。结果CPP测试显示,实验组、模型组和正常组在伴药箱的停留时间分别为(246.70±9.28)、(457.90±12.20)和(206.80±9.45)s,运动路程分别为(3536.90±110.54)、(4436.10±115.49)和(3410.40±94.54)cm;模型组在伴药箱的停留时间及运动路程与正常组和实验组比较,差异均有统计学意义(均P<0.01)。蛋白质印迹结果显示,外泌体特异性表达TSG101和CD63。基因芯片结果显示,甲基苯丙胺可上调276个血清外泌体miRNAs表达,其中50个差异均有统计学意义(均P<0.05),可下调25个miRNAs表达,其中2个差异均有统计学意义(均P<0.05);钩藤碱可下调228个miRNAs表达,其中32个差异均有统计学意义(均P<0.05),可上调51个miRNAs表达,其中9个差异均有统计学意义(均P<0.05)。有189个miRNAs经甲基苯丙胺诱导后表达上调,但经钩藤碱治疗后出现下调。结论差异表达的血清外泌体miRNAs可能是甲基苯丙胺成瘾的重要原因之一,钩藤碱可以明显减轻甲基苯丙胺引起的大鼠位置偏爱及血清外泌体miRNAs表达的改变。Objective To observe the effects of rhynchophylline on conditioned place preference and expression of serum exosome miRNAs in methamphetamine-dependent rats.Methods Conditioned place preference(CPP)rat model was established by methamphetamine.30 rats were randomly divided into three groups:normal group,model group and experimental group.2 mg·kg^(-1) methamphetamine was used for modeling in model and experimental groups by subcutaneous injection,and only 0.9%NaCl was injected in normal group,while 60 mg·kg^(-1) rhynchophylline was used for intervention in experimental group by intraperitoneal injection.The expression of biomarker in serum exosomes was determined by Western blot.The expression profile of miRNAs in ser um exosomes was determined by gene chip.The expression of miRNAs was calculated and corrected by sRNA bench software,and miRNAs expressed differentially between groups were analyzed by sugar-edge study software.Results The CPP test showed that the residence time in the companion box was(246.70±9.28),(457.90±12.20)and(206.80±9.45)s;the distance of movement was(3536.90±110.54),(4436.10±115.49)and(3410.40±94.54)cm for experimental group,model group and normal group,respectively.Compared with normal group and experimental group,model group had statistically significant differences of residence time and moving distance in the companion box(all P<0.01).Western blot showed that TSG101 and CD63 were specifically expressed in exosomes.The analysis of serum exosome miRNAs expression profiles indicated that methamphetamine could up-regulate 276 miRNAs expressions,of which 50 were statistically different(all P<0.05),and also down-regulate 25 miRNAs,among which 2 were statistically significant(all P<0.05).Rhynchophylline could down-regulate 228 miRNAs,of which 32 were statistically different(all P<0.05),and also up-regulate 51 miRNAs,among which 9 were statistically different(all P<0.05).189 miRNAs were up-regulated after induction by methamphetamine and down-regulated after treatment with rhynchophylline.

关 键 词：钩藤碱 甲基苯丙胺 药物依赖 中药神经药理 血清外泌体 miRNA 

分 类 号：R28[医药卫生—中药学]