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目的验证通过生物信息学技术筛选出Yes-相关蛋白(YAP)在大鼠心肌缺血再灌注损伤中的表达情况,为后期进一步研究奠定基础。方法利用生物信息学技术分析大鼠心肌缺血再灌注损伤的差异基因,应用KOBAS2.0以及KEGG数据库筛选出相关的信号通路和关键基因。将18只Sprague Dawley大鼠随机分配为正常组(n=6)、假手术组(n=6)、缺血再灌注损伤组(模型组,n=6),采用免疫组织化学、逆转录荧光定量聚合酶链反应、蛋白质印迹法检测筛选出的靶基因的表达情况。结果通过生物信息学技术筛选出差异表达基因共345条,上调181条,下调164条;筛选出的差异基因主要富集于Wnt、HIPPO、MAKP、Jak-STAT等信号通路;该课题组关注HIPPO信号通路,发现与正常组和假手术组比较,模型组中其关键蛋白YAP在基因以及蛋白表达水平增加,差异有统计学意义(P<0.05)。结论经生物信息学筛选出来的HIPPO信号通路关键蛋白YAP在大鼠心肌缺血再灌注损伤中表达增加。
Objective To verify the expression of Yes-related protein (YAP) in myocardial ischemia-reperfusion injury in rats through bioinformatics technology and lay the foundation for further study. Methods Bioinformatics techniques were used to analyze the differentially expressed genes in myocardial ischemia-reperfusion injury in rats. KOBAS2.0 and KEGG databases were used to screen out the related signaling pathways and key genes. Eighteen Sprague Dawley rats were randomly divided into normal group (n = 6), sham operation group (n = 6) and ischemia - reperfusion injury group (model group, n = 6). Immunohistochemistry and reverse transcription fluorescence Quantitative polymerase chain reaction and Western blotting were used to detect the expression of target genes. Results A total of 345 differentially expressed genes were screened out by bioinformatics techniques, with 181 up-regulated and 164 down-regulated genes. The selected genes were mainly enriched in Wnt, HIPPO, MAKP and Jak-STAT signaling pathways. The group focused on HIPPO Signal pathways. Compared with the normal group and sham operation group, the expression of YAP, a key protein in the model group, increased in gene and protein levels, with statistical significance (P <0.05). Conclusion YAP, a key protein of HIPPO signaling pathway, has been screened out by bioinformatics in myocardial ischemia-reperfusion injury in rats.