VSMC源KLF5对腹主动脉瘤形成中作用及机制的研究  

The Role of VSMCs-derived KLF5 in Abdominal Aortic Aneurysm Formation and the Underlying Mechanisms

作　　者：孟丽 朱艳 徐彦楠[2] 马冬 周晨明 MENG Li;ZHU Yan;XU Yannan(Electron Microscopy Center,Hebei Medical University,Shijiazhuang City,Hebei Province 050017;不详)

机构地区：[1]河北医科大学电镜实验中心,河北省石家庄市050017 [2]河北医科大学教学实验中心,河北省石家庄市050017 [3]华北理工大学公共卫生学院

出　　处：《医学理论与实践》2022年第12期1981-1984,1989,共5页The Journal of Medical Theory and Practice

基　　金：国家自然科学基金资助项目(81541149)。

摘　　要：目的:应用血管平滑肌细胞特异性敲除KLF5基因小鼠(VSMC^(KLF5-/-))、基因芯片和生物信息学技术,探讨VSMC源KLF5在腹主动脉瘤(AAA)形成中的作用及机制。方法:通过CaPO_(4)构建VSMC^(KLF5-/-)与野生型小鼠(WT)腹主动脉瘤模型,采用HE染色检测VSMC中KLF5基因缺失对AAA膨胀变化影响;通过RNA的提取及mRNA表达谱的检测对mRNA表达谱进行分析;采用差异表达基因的富集分析探讨VSMC中KLF5调节差异表达基因在AAA形成中的生物学功能;采用Western blot检测Stk3、Rassf4、Wwtr1蛋白表达。结果:HE染色显示两组小鼠腹主动脉管壁变薄,且VSMC^(KLF5-/-)组血管膨胀更为明显。mRNA芯片筛查结果显示,与WT组比较,VSMC^(KLF5-/-)组血管组织中共上调表达基因479个,下调基因514个;GO分析显示上调基因主要富集在转录共激活因子活性、细胞骨架重塑等,下调基因富集在前体代谢产物和能量的产生。KEGG信号通路分析显示上调基因主要富集在Hippo信号通路,下调基因富集于细胞能量代谢相关信号通路。Western blot验证VSMC^(KLF5-/-)促进CaPO_(4)诱导血管组织中Stk3、Rassf4、Wwtr1的表达升高。结论:血管平滑肌细胞中转录因子KLF5可能通过调控Hippo通路维持血管稳态并抑制AAA的形成。Objective:To investigate the role and mechanism of VSMC-derived KLF5 in the formation of abdominal aortic aneurysm(AAA)in vascular smooth muscle cell-specific knockout mice(VSMC^(KLF5-/-)),gene chip and bioinformatics techniques.Methods:VSMC^(KLF5-/-) and wild-type mouse(WT)abdominal aortic aneurysm models were constructed by CaPO_(4).The influence of KLF5 gene deletion on AAA swelling in VSMC was detected by HE staining.The mRNA expression profile was analyzed by RNA extraction and mRNA expression profile detection.The biological function of KLF5-regulated genes in the formation of AAA in VSMC was investigated by enrichment analysis of differentially expressed genes.The protein expressions of Stk3,Rassf4 and Wwtr1 were detected by Western blot.Results:HE staining showed the pathological changes of abdominal aortic wall thinning in the two groups,the VSMC^(KLF5-/-) group was more obvious.The results of mRNA chip screening showed that compared with WT group,479 up-regulated genes and 514 down-regulated genes were expressed in vascular tissues of VSMC^(KLF5-/-) group.GO analysis showed that up-regulated gene were mainly concentrated in chromosome segregation,transcription coactivator activity and microtubule plus-end;down-regulated genes were concentrated in generation of precursor metabolites and energy,cofactor binding,sarcomere and other aspects.Analysis of KEGG pathway showed that up-regulated genes were mainly concentrated in Hippo pathway,while down-regulated genes were concentrated in cell energy metabolism related signaling pathway.Western blot assay confirmed that VSMCs-specific deletion of KLF5 in AAA tissues promotes the expression of Stk3,Rassf4 and Wwtr1.Conclusion:The transcription factor KLF5 in vascular smooth muscle cells may maintain vascular homeostasis and inhibit the formation of AAA by regulating Hippo pathway.

关 键 词：腹主动脉瘤 血管平滑肌 KLF5 生物信息学 Hippo通路 

分 类 号：R34[医药卫生—基础医学]