白花香莲解毒颗粒抑制HBV的miRNA表达谱筛选  

Screening of microRNAs expression profile of Baihua Xianglian Jiedu Granule in inhibiting HBV

作　　者：黄鹏 邱华[2] 刘雪梅[1] 范春娇 李家焕 牙程玉 覃秀容 胡晓 吴聪[1] HUANG Peng;QIU Hua;LIU Xue-mei;FAN Chun-jiao;LI Jia-huan;YA Cheng-yu;QIN Xiu-rong;HU Xiao;WU Cong(Guangxi University of Chinese Medicine,Nanning Guangxi 530001,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning Guangxi 530023,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《时珍国医国药》2020年第7期1551-1555,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81660827;81860889);中国博士后科学基金面上资助项目(2019M653312);广西重点研发计划项目(2017AB45166);广西中医药大学研究生教育创新计划项目(xjyb066);广西中药制剂共性技术研发重点实验室项目(ZJGX2016006;ZJGX201502001);第二批广西高层次骨干人才培养139计划(桂卫科教发[2018]22号)

摘　　要：目的筛选出白花香莲解毒颗粒抑制乙型肝炎病毒(HBV)复制的miRNA表达谱,旨在发现及鉴定新的具有抗HBV活性的miRNA。方法采用miRNA芯片和基因芯片技术检测HepG2细胞空白对照组、HBV 1.3P侵染HepG2细胞模型组和白花香莲解毒颗粒干预组的表达谱,通过TargetScan以及miRDB数据库预测miRNA的靶基因,并对靶基因进行GO和KEGG Pathways分析,用qRT-PCR方法对筛选出的miRNA进行验证。结果 miRNA芯片筛选出与白花香莲解毒颗粒抑制HBV相关的20个发生显著变化的miRNA,其中在模型组表达降低,同时在白花香莲解毒颗粒干预组表达升高的有16个;在模型组表达升高,同时在白花香莲解毒颗粒干预组表达降低的有4个;qRT-PCR验证hsa-miR-106a-5p、hsa-miR-181a-3p、hsa-miR-20a-5p、hsa-miR-141-3p以及hsa-miR-372-3p的差异表达,结果与芯片一致。结论白花香莲解毒颗粒对肝细胞内的miRNA表达具有干预作用,能通过调控miRNA发挥抑制HBV的作用。Objective To screen out the expression profiles of microRNAs inhibiting hepatitis B virus(HBV) replication by Baihua Xianglian Jiedu Granule in order to discover and identify novel microRNAs with anti-HBV activity. Methods The expression profiles of HepG2 cells in blank control group, HBV1.3-D infected HepG2 cell model group and Baihua Xianglian Jiedu Granule intervention group were detected by microarray and gene chip technology. Target Scan and microarray DB databases were used to predict the target genes of microRNAs. GO and KEGG Pathways were used to analyze the target genes, and qRT-PCR was used to screen them. The selected microRNAs were validated.Results Twenty significantly altered microRNAs related to the inhibition of HBV by Baihua Xianglian Jiedu Granule were screened by microarray, including 16 in the model group and 16 in the intervention group of Baihua Xianglian Jiedu Granule, and 4 in the model group and the intervention group of Baihua Xianglian Jiedu Granule. QRT-PCR was used to verify the differential expression of hsa-mir-106 a-5 p, has-mir-181 a-3 p, hsa-mir-20 a-5 p, hsa-mir-141-3 p and hsa-mir-372-3 p. The results were consistent with the chip. Conclusion Baihua Xianglian Jiedu Granule can interfere with the expression of microRNAs in hepatocytes and inhibit HBV by regulating the expression of microRNAs.

关 键 词：壮医药 白花香莲解毒颗粒 乙型肝炎病毒 微小RNA 复制 表达谱 

分 类 号：R284.1[医药卫生—中药学]