三种中药活性成分对吗啡依赖斑马鱼脑中dre-miR-723-5p表达的影响  

Effects of three kinds of active ingredients of traditional Chinese medicine on dre-miR-723-5p expression in morphine-induced zebrafish brain

作　　者：朱晨 林汝堃 区炳雄 陈志杰[1] 罗超华[1] 莫志贤[1] 李璟 ZHU chen;LIN Ru-kun;OU Bing-xiong;CHEN Zhi-jie;LUO Chao-hua;MO Zhi-xian;LI Jing(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510515,China;Central Laboratory,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学中医药学院,广东广州510515 [2]广州中医药大学第二附属医院,广东广州510515 [3]南方医科大学中心实验室,广东广州510515

出　　处：《中国药理学通报》2021年第11期1542-1547,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81873030);广州市科技计划项目(No201707010287);广东省基础与应用基础研究基金-自然科学基金项目(No2021A1515011565)。

摘　　要：目的研究三种中药活性成分(青藤碱、钩藤碱和异钩藤碱)对吗啡依赖斑马鱼脑中dre-miR-723-5p表达的影响。方法斑马鱼腹腔注射吗啡后,进行条件性位置偏爱(CPP)训练,观察行为学;采用小RNA测序检测斑马鱼脑区miRNA表达谱;qRT-PCR验证差异表达量最大的dre-miR-723-5p并对其进行靶基因预测、GO和KEGG通路分析。结果与对照组相比,模型组斑马鱼在伴药箱中的停留时间和运动轨迹明显增加,CPP模型成功建立;给予药物处理后得以改善;qRT-PCR验证结果与小RNA测序结果一致;预测dre-miR-723-5p共同的靶基因有99个,主要富集在生物过程、细胞组成和分子功能,参与了MAPK信号通路、溶酶体、细胞因子-细胞因子受体相互作用和细胞凋亡等方面。结论三种中药活性成分可以拮抗吗啡依赖斑马鱼CPP效应和dre-miR-723-5p表达变化,dre-miR-723-5p可能与吗啡依赖形成有关。Aim To study the effect of three kinds of active ingredients of traditional Chinese medicine(sinomenine,rhynchophylline and isorhynchophylline)on dre-miR-723-5p expression in morphine-induced zebrafish brain.Methods Morphine was injected intraperitoneally to zebrafish,conditional position preference(CPP)was trained and then the behavioral of animals were observed;the miRNA expression profiles of morphine-additive zebrafish were determined by small RNA sequencing;qRT-PCR was used to verify the expression of dre-miR-723-5p,three target gene databases(miRanda,miRDB,andRNAhybrid)were used to predict the target genes of dre-miR-723-5p;Kobas 3.0 was used to perform Gene Ontology(GO)and KEGG pathway analysis of these target genes.Results Morphine-induced CPP model was established successfully.Compared with control group,the resident time and movement map in drug-pair box of zebrafish in model group significantly increased.After drug administration,the resident time and movement map in drug-pair box of zebrafish decreased.The verification results of qRT-PCR were consistent with the results of small RNA sequencing.Ninety-nine putative target genes of dre-miR-723-5p that were common to all three target gene databases,which were mainly enriched in biological process,cell composition and molecular function,involved in the positive regulation of MAPK signaling pathway,lysosome,cytokine-cytokine receptor interaction,and apoptosis.Conclusion Morphine can increase the expression of dre-miR-723-5p in the zebrafish brain,which can be reversed by sinomenine,isorhynchophylline,and rhynchophylline treatment,and dre-miR-723-5p may participate in the mechanism underlying morphine-induced damage of brain.

关 键 词：吗啡 dre-miR-723-5p 青藤碱 钩藤碱 异钩藤碱 斑马鱼 

分 类 号：R-332[医药卫生] R284.1