摘要
背景腹主动脉瘤(AAA)是一种老年常见高死亡率的心血管系统疾病,发病机制不明确,临床上无有效治疗药物。本课题组前期研究发现,KLF5基因在人和小鼠AAA组织中表达升高,但是对于详细的KLF5基因调控炎症相关基因表达的网络分析未见报道。目的利用基因芯片技术对髓系KLF5基因表达缺失(LyKLF5-/-)小鼠AAA的差异表达基因进行生物信息学分析,探讨KLF5基因在AAA中的调控机制。方法研究时间为2015年1月-2019年4月。将SPF级KLF5-flox小鼠和SPF级LysM-cre小鼠杂交培育的雄性髓系遗传性LyKLF5-/-小鼠与同笼野生型(WT)C57BL/6小鼠各3只作为实验动物。通过CaPO4诱导小鼠AAA模型,并提取肾下腹主动脉组织总RNA行m RNA芯片筛查。对于筛选出的差异表达基因,利用在线数据库Metascape进行GO功能富集分析和KEGG富集分析,再利用STRING在线数据库进行蛋白互作网络分析。结果 mRNA芯片共筛选到上调表达基因646个,下调表达基因1 410个(|FC|≥2,P<0.05)。与WT小鼠比较,抑制炎症的基因在LyKLF5-/-小鼠表达增加,Bcas3、Hck基因表达分别上调3.96、3.14倍;促进炎症的基因在LyKLF5-/-小鼠表达降低,Retnla、Olig1、Tnfrsf11a分别下调7.68、5.07、3.66倍。GO功能富集分析显示上调差异基因主要参与多肽抗原合成和表达的调控、核苷二磷酸代谢过程、肌节组织的正向调节等过程;下调差异基因主要参与生长发育、细胞对葡萄糖刺激反应的胰岛素分泌调节、平滑肌细胞迁移的正调控等过程。KEGG富集分析显示上调差异基因主要影响破骨细胞分化、吞噬小体、氨基酸的生物合成等通路;下调差异基因主要影响血管平滑肌收缩、泛素介导的蛋白水解作用以及肌动蛋白细胞骨架的调控等通路。蛋白互作网络分析证实,小鼠髓系LyKLF5-/-导致参与血管炎症相关基因Bcas3、Hck表达上调和Retnla、Olig1、Tnfrsf11a表达下调。结论 Bcas3、Hck、Retnla
Background Abdominal aortic aneurysm(AAA) is a common cardiovascular disease with high mortality in the elderly.Its pathogenesis remains unclear and there is no special effective pharmacological treatment.Our previous research found increased KLF5 expression in human and mouse AAA tissues,but a detailed network analysis of KLF5 regulating inflammatory gene expression is still unreported.Objective To investigate the regulating role of KLF5 in AAA by bioinformatically analyzing the aberrant expressed genes in CaPO4-injured AAA tissues from LyKLF5-/-and WT mice using gene chip technology.Methods Between January 2015 and April 2019,SPF grade KLF5-flox mice and SPF grade LysMcre mice were cross-bred to produce male mice with LyKLF5-/-and WT C57 BL/6,respectively,each group for 3 mice.Extraction of total RNA in CaPO4-injured abdominal aorta from AAA model was used for mRNA microarray screening.The ectopic gene expression was analyzed by functional annotation(GO) and KEGG.Protein interaction network analysis was conducted by STRING.Results A total of 646 up-regulated genes and 1 410 down-regulated genes were detected(|FC| ≥ 2,P<0.05).Compared with WT group,the expression of anti-inflammatory genes was significantly increased in LyKLF5-/-group,Bcas3 and Hck genes were obviously up-regulated about 3.96 and 3.14 times,respectively.On the other hand,among the decreased proinflammatory genes,Retnla,Olig1 and Tnfrsf11 a were obviously down-regulated by 7.68,5.07 and 3.66 times.GO analysis showed that up-regulated genes were mainly enriched in the regulation of polypeptide antigen processing and presentation,the nucleoside diphosphate metabolic process,and the positive regulation of sarcomere organization.Down-regulated genes were enriched in the development and growth,regulation of glucose-stimulated insulin secretion and smooth muscle cell migration.KEGG signal pathway analysis showed that up-regulated differential genes mainly affected osteoclast differentiation,phagosome,biosynthesis of amino acids and other pathways.Dow
作者
常全
张娜娜
马冬
CHANG Quan;ZHANG Nana;MA Dong(School of Public Health,North China University of Science and Technology,Tangshan 063210,China)
出处
《中国全科医学》
CAS
北大核心
2020年第6期692-698,共7页
Chinese General Practice
基金
国家自然科学基金资助项目（81541149）
关键词
主动脉瘤
腹
髓系细胞
KLF5基因
表达谱芯片
小鼠
Aortic aneurysm,abdominal
Myeloid cells
KLF5 gene
Expression spectrum chip
Mice
