SIRT1对CCM内皮细胞EndMT及细胞增殖和迁移的调节作用  

Regulatory effect of SIRT1 on EndMT and cell proliferation,migration of endothelial cells of cerebral cavernous malformations

作　　者：曹沅 熊子瑜 李莹[1] 闫聪[1] 王煜文 门春阳 叶永青 毕锐 何志怡 段前鹏 杜家培 呼斯楞图 石长斌[1] CAO Yuan;XIONG Ziyu;LI Ying;YAN Cong;WANG Yuwen;MEN Chunyang;YE Yongqing;BI Rui;HE Zhiyi;DUAN Qianpeng;DU Jiapei;HU Silengtu;SHI Changbin(Department of Neurosurgery,First Affiliated Hospital of Harbin Medical University,Harbin 150007,China)

机构地区：[1]哈尔滨医科大学附属第一医院神经外科,哈尔滨150007

出　　处：《山西医科大学学报》2025年第3期267-274,共8页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(82071298)。

摘　　要：目的探讨沉默信息调节因子2同源物1(SIRT1)对脑海绵状血管畸形(CCM)内皮细胞中内皮-间充质转化(EndMT)相关转录因子及细胞表型的影响。方法免疫荧光染色法检测正常脑组织和CCM病变组织血管内皮细胞SIRT1的表达。慢病毒转染方法敲低人脑微血管内皮细胞(hCMEC)中的PDCD10基因,构建CCM细胞模型。用SIRT1激动剂SRT-1720分别处理正常内皮细胞和CCM内皮细胞48 h,设立4个实验组:shBlank组、shBlank+SRT-1720组、shPDCD10组、shPDCD10+SRT-1720组。qPCR检测内皮细胞PDCD10敲低效率、SIRT1及EndMT相关转录因子(SNAI1、SNAI2、SNAI3、TWIST1、TWIST2)表达水平;Western blot分析PDCD10敲低效率及SIRT1的蛋白表达;EdU细胞增殖实验评估细胞增殖情况;细胞划痕实验检测细胞迁移能力。结果与正常脑组织相比,CCM病变组织中的内皮细胞SIRT1表达下调。与shBlank组比较,shPDCD10组PDCD10和SIRT1的mRNA及蛋白水平显著降低(P<0.05),SNAI1、SNAI2、SNAI3、TWIST1的mRNA水平显著升高(P<0.05),TWIST2的mRNA水平显著下降(P<0.05),细胞增殖和迁移能力显著增强(P<0.05)。与shPDCD10组相比,shPDCD10+SRT-1720组SIRT1的mRNA及蛋白水平显著上调(P<0.05),SNAI2、SNAI3、TWIST1的mRNA水平显著降低(P<0.05),TWIST2的mRNA水平显著升高(P<0.05),细胞增殖和迁移能力显著减弱(P<0.05)。结论SIRT1抑制CCM内皮细胞中EndMT相关转录因子异常表达,减缓CCM内皮细胞的异常增殖和迁移,提示SIRT1可能成为治疗CCM的新靶点。Objective To explore the effect of silent information regulator 1(SIRT1)on endothelial-mesenchymal transition(EndMT)-related transcription factors and cell phenotypes in endothelial cells of cerebral cavernous malformations(CCM).Methods Immunofluorescence staining was used to detect the expression of SIRT1 in vascular endothelial cells of normal brain tissue and CCM lesion tissue.Lentiviral transfection was used to knock down the PDCD10 gene in human cerebral microvascular endothelial cells(hCMEC)to establish a CCM cell model.SIRT1 agonist SRT-1720 was used to treat normal endothelial cells and CCM endothelial cells for 48 h,named as shBlank group,shBlank+SRT-1720 group,shPDCD10 group,and shPDCD10+SRT-1720 group,respectively.qPCR was performed to measure the knockdown efficiency of PDCD10,and the expression levels of SIRT1 and EndMT-related transcription factors(SNAI1,SNAI2,SNAI3,TWIST1,TWIST2).Western blot was used to analyze PDCD10 knockdown efficiency and protein expression of SIRT1.EdU assay was used to evaluate the cell proliferation.Cell scratch assay was used to test the cell migration.Results Compared with normal brain tissue,the expression of SIRT1 in endothelial cells of CCM lesion tissue was downregulated.Compared with shBlank group,the mRNA and protein levels of PDCD10 and SIRT1 were significantly reduced in shPDCD10 group(P<0.05),the mRNA levels of SNAI1,SNAI2,SNAI3,TWIST1 were significantly increased(P<0.05),the mRNA level of TWIST2 was significantly decreased(P<0.05),and the cell proliferation and migration abilities were significantly enhanced(P<0.05).Compared with shPDCD10 group,the mRNA and protein levels of SIRT1 were significantly upregulated in shPDCD10+SRT-1720 group(P<0.05),the mRNA levels of SNAI2,SNAI3,TWIST1 were significantly reduced(P<0.05),the mRNA level of TWIST2 was significantly increased(P<0.05),and the cell proliferation and migration abilities were significantly weakened(P<0.05).Conclusion SIRT1 can inhibit the abnormal expression of EndMT-related transcription factors in CCM en

关 键 词：脑海绵状血管畸形 SIRT1 内皮细胞 内皮-间充质转化 细胞增殖 细胞迁移 

分 类 号：R743[医药卫生—神经病学与精神病学]