采用基因芯片技术研究丹红注射液对急性心肌梗死模型大鼠基因表达谱的影响
x

请在关注微信后,向客服人员索取文件

篇名: 采用基因芯片技术研究丹红注射液对急性心肌梗死模型大鼠基因表达谱的影响
TITLE:
摘要: 目的:探讨丹红注射液(DHI)对急性心肌梗死(AMI)模型大鼠基因表达谱的影响。方法:将雄性SD大鼠随机分为假手术组、模型组和DHI组(0.76 mL/kg),每组10只。模型组和DHI组采用冠状动脉左前降支结扎法复制AMI模型。造模后,假手术组和模型组大鼠均肌内注射等容生理盐水,DHI组大鼠肌内注射相应药物,每日1次,连续14 d。末次给药后,分离大鼠梗死边缘区心肌组织,采用基因芯片技术检测基因表达谱的变化情况,以相对表达量差异倍数为指标,筛选差异表达微RNA(miRNA)。在检索其对应基因的基础上,利用DAVID生物信息学资源数据库和KEGG通路数据库分别进行基因本体(GO)和KEGG通路富集分析;借助TargetScan数据库预测差异表达miRNA对应的靶基因信使RNA(mRNA),采用Cytoscape 3.6.1软件构建miRNA- mRNA网络并进行分析,采用Agilent GeneSpring GX v11.5软件筛选上述网络中与炎症相关的靶基因和miRNA。结果:与假手术组比较,模型组差异表达miRNA共22个,其中上调5个、下调17个;与模型组比较,DHI组差异表达miRNA共26个,均为上调;与DHI治疗AMI有关的差异表达miRNA包括rno-let-7a-5p、rno-let-7d-5p、rno-let-7f-5p、rno-miR-26b-5p、rno-miR-29b-3p、cel-miR- 39-3p、cel-miR-39-5p、rno-miR-142-5p、rno-miR-191a-5p、rno-miR-409a-3p。GO和KEGG通路富集分析结果显示,差异表达miRNA对应基因主要集中在膜结合细胞器、细胞质、内膜系统等细胞组分中,通过解剖结构发育、多细胞组织发育、发育过程等生物过程来发挥蛋白结合、离子结合等分子功能;其主要富集于钙信号通路,过氧化物酶体增殖物激活受体(PPAR)信号通路,血管内皮生长因子(VEGF)信号通路,细胞凋亡,糖基磷脂酰肌醇锚定生物合成,缬氨酸、亮氨酸和异亮氨酸降解等信号通路上。miRNA-mRNA网络分析结果显示,与差异表达miRNA对应的靶基因mRNA共25个,与之关联的miRNA共24个;该网络中与炎症相关的靶基因共6个(IL6、IL1b、TNF、TLR4、CRP、CXCL12),涉及差异表达miRNA共19个。结论:DHI对AMI的治疗作用可能与调节相关miRNA的表达,影响钙离子、PPAR、VEGF等通路的信号转导,调控白细胞介素、趋化因子、C反应蛋白等炎症标志物的分泌有关。
ABSTRACT: OBJECTIVE: To investigate the effects of Danhong injection (DHI) on gene expression profile of acute myocardial infarction (AMI) model rats. METHODS: Male SD rats were randomly divided into sham operation group, model group and DHI group (0.76 mL/kg), with 10 rats in each group. AMI model was established by ligation of left anterior descending coronary artery in model group and DHI group. After modeling, sham operation group and model group were given constant volume of normal saline intramuscularly, and DHI group was given relevant medicine intramuscularly, once a day, for consecutive 14 days. After last administration, myocardial tissue in the marginal zone of infarction was separated. The change of gene expression profile was detected by gene chip technique. Using fold-change of relative expression as index, differentially expressed microRNA (miRNA) were screened. On the basis of retrieving their corresponding genes, gene ontology (GO) and KEGG pathway enrichment analysis were carried out by using DAVID bioinformatics resource database and KEGG pathway database, respectively. TargetScan database was used to predict the target gene messenger RNA (mRNA) corresponding to differentially expressed miRNA. Cytoscape 3.6.1 software was used to construct and analyze the miRNA-mRNA network. Agilent GeneSpring GX v11.5 software was used to screen target genes and miRNA related to inflammation in the above networks. RESULTS: Compared with sham operation group, there were 22 differentially expressed miRNAs in model group, 5 up-regulated and 17 down-regulated. Compared with model group, there were 26 differentially expressed miRNAs in DHI group, and all of them were up-regulated. The differentially expressed miRNAs related to DHI therapy for AMI included rno-let-7a-5p, rno-let-7d-5p, rno-let-7f-5p, rno-miR-26b-5p, rno-miR-29b-3p, cel-miR-39-3p, cel-miR-39-5p, rno-miR-142-5p, rno-miR-191a-5p, rno-miR-409a-3p. Results of GO analysis and KEGG pathway enrichment analysis showed that differentially expressed miRNAs were mainly concentrated in membrane-bound organelles, cytoplasm, endometrial system and other cell components. The molecular functions such as protein binding and ion binding were exerted through biological processes such as anatomical structure development, multicellular tissue development and development process,which were mainly enriched in calcium signaling pathway, PPAR signaling pathway, VEGF signaling pathway, cell apoptosis, glycosylphosphatidylinositol anchored biosynthesis, valine, leucine and isoleucine degradation, etc. miRNA-mRNA network analysis showed that there were 25 target gene mRNAs corresponding to differentially expressed miRNA and 24 miRNAs related to it. There were 6 inflammation-related target genes (IL6, IL1b, TNF, TLR4, CRP, CXCL12) in this network, involving 19 differentially expressed miRNAs. CONCLUSIONS: The therapeutic effect of DHI on AMI may be related to regulating the expression of related miRNA, affecting signal transduction of calcium ion, PPAR and VEGF pathways, and regulating the secretion of inflammatory markers such as interleukin, chemokine and C-reactive protein.
期刊: 2019年第30卷第22期
作者: 施洋,樊官伟,候宝林,樊登峰,张伟,鲁西亮,陈晓黎,何敏
AUTHORS: SHI Yang,FAN Guanwei,HOU Baolin,FAN Dengfeng,ZHANG Wei,LU Xiliang,CHEN Xiaoli,HE Min
关键字: 基因芯片技术;丹红注射液;急性心肌梗死;差异表达;微RNA;信使RNA;信号通路;炎症;大鼠
KEYWORDS: Gene chip technique; Danhong injection; Acute myocardial infarction; Differential expression; MicroRNA; Messenger RNA; Signaling pathway; Inflammation; Rat
阅读数: 75 次
本月下载数: 4 次

* 注:未经本站明确许可,任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作!