芯片技术筛选夹脊穴预处理心肌缺血再灌注大鼠Wnt通路相关差异表达基因

发布时间：2018-08-14 18:41

【摘要】：目的:通过基因芯片技术筛选心肌缺血再灌注损伤(MIRI)大鼠差异表达的基因,富集分析关键通路,寻找电针夹脊穴预处理可能的保护途径。方法:72只大鼠,随机分为对照组、模型组、夹脊组、内关组、阳陵泉组、曲池组,每组12只。各针刺组电针预处理7 d后,制备MIRI模型。提取缺血区心肌组织总RNA并评估质量后,进行样品标记和芯片杂交,使用Agilent Feature Extraction软件获得芯片图,Gene Spring GX v12.1软件进行Quantile标准化及数据处理,使用标准的富集计算方法进行GO分析和Pathway分析。并采用荧光定量PCR对筛选出的差异基因进行部分验证。结果:电针预处理能够调控缺血再灌注大鼠的基因表达,其中夹脊穴组对Wnt通路相关基因影响显著,共涉及10个基因:上调细胞外因子Wnt9b、Wnt10b,蛋白受体Fzd2,磷脂酶C家族Plch1、plch2,DKK家族中Dkkl1及APC家族基因Amer1和Amer3;同时下调磷脂酶C家族Pclb1及Sapcd家族基因Sapcd2。这些基因在内关、阳陵泉、曲池组无显著性差异。PCR结果与芯片结果相符。结论:多条信号通路参与电针夹脊穴预处理对心肌缺血再灌注的调控,涉及多个基因。从分子水平阐明了其对MIRI的保护作用机制。
[Abstract]:Objective: to screen differentially expressed genes in (MIRI) rats with myocardial ischemia-reperfusion injury by gene chip technique, to enrich and analyze the key pathways, and to find out the possible protective pathway of electroacupuncture Jiaji point preconditioning. Methods Seventy-two rats were randomly divided into control group, model group, Jiaji group, Neiguan group, Yanglingquan group and Quchi group with 12 rats in each group. The MIRI model was prepared by electroacupuncture pretreatment for 7 days in each acupuncture group. After total RNA was extracted from myocardial tissue of ischemic area and the quality was evaluated, sample labeling and chip hybridization were performed, and gene Spring GX v12.1 software was used to obtain Agilent Feature Extraction software for Quantile standardization and data processing. Go analysis and Pathway analysis were carried out using the standard enrichment calculation method. The differentially screened genes were partially verified by fluorescence quantitative PCR. Results: electroacupuncture preconditioning could regulate the gene expression in ischemia-reperfusion rats, and Jiaji acupoint group had a significant effect on the genes related to Wnt pathway. There were 10 genes involved: up-regulation of the extracellular factor Wnt9bT10b, protein receptor Fzd2, Dkkl1 and APC family genes Amer1 and Amer3 in the Plch1plch2DKK family of phospholipase C, and down-regulation of Pclb1 gene of phospholipase C family and Sapcd2 gene of Sapcd family at the same time. There was no significant difference between these genes in Neiguan, Yanglingquan and Quchi groups. PCR results were consistent with microarray results. Conclusion: multiple signal pathways are involved in the regulation of myocardial ischemia reperfusion by electroacupuncture Jiaji point preconditioning. The mechanism of its protective effect on MIRI was elucidated at molecular level.
【作者单位】：上海市第七人民医院;山东中医药大学针灸推拿学院;山东中医药大学附属医院针灸科;
【基金】：国家自然科学基金项目(81373722) 山东省自然科学基金项目(ZR2014HL100)
【分类号】：R245

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