CKAP2对高糖培养的人视网膜血管内皮细胞的作用研究  

Effect of CKAP2 on Human Retinal Vascular Endothelial Cells Cultured in High Glucose and Its Mechanism

作　　者：陈湘婷 谢洁 孟倩丽 CHEN Xiang-ting;XIE Jie;MENG Qian-li(School of Medicine,South China University of Technology,Guangzhou 510006,China;Department of Ophthalmology,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangdong Eye Institute,Guangzhou 510080,China;The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510400,China)

机构地区：[1]华南理工大学医学院,广州510006 [2]广东省人民医院眼科、广东省医学科学院、广东省眼病防治研究所,广州510080 [3]南方医科大学第二临床学院,广州510400

出　　处：《循证医学》2020年第2期96-102,共7页The Journal of Evidence-Based Medicine

基　　金：广东省医学科学技术研究基金资助项目(A2019380,A2018401);广东省自然科学基金资助项目(2017A030313609,2019A1515010697)。

摘　　要：目的探讨细胞骨架蛋白2(CKAP2)对高糖培养的人视网膜血管内皮细胞的作用。方法体外培养人视网膜微血管内皮细胞(HRMECs)株,分为正常对照组(CON组)、高渗培养组(30 mmol/L甘露醇,HO组)、高糖培养组(30 mmol/L D-葡萄糖,HG组)。RT-PCR和Western blot法分别检测CKAP2 mRNA及蛋白表达情况。分别用siCKAP2转染HRMECs阻断CKAP2,或加入雷珠单抗(106μg/mL)阻断血管内皮生长因子后,细胞增殖、划痕和小管形成实验检测细胞的增殖率、迁移能力和成管能力。结果①与正常对照组、高渗组相比,高糖组CKAP2的mRNA和蛋白表达上调;②阻断CKAP2后,细胞的增殖能力、迁移能力和血管形成能力明显减弱;③阻断CKAP2与阻断血管内皮生长因子对降低细胞的增殖、迁移和成管能力的作用相似。结论 CKAP2可影响视网膜微血管内皮细胞的增殖、迁移和成管能力,其作用效果可能与血管内皮生长因子相似。Objective To investigate the role of cytoskeletal-associated protein 2(CKAP2)in human retinal vascular endothelial cells cultured in high glucose.Methods Human retinal microvascular endothelial cells(HRMECs)were cultured in vitro and divided into normal control group(CON group),hypertonic culture group(30 mmol/L mannitol,HO group),and high glucose culture group(30 mmol/L D-glucose,HG group).Real-time PCR and Western blot were used for detecting the mRNA and protein expression level of CKAP2.After transfection of HRMECs with siCKAP2 to block CKAP2,or addition of Ranibizumab(106μg/mL)to block VEGF,MTT,scratch and tubule formation experiments were performed to detect the proliferation rate,migration ability and tube formation ability of cells.Results①Compared with the CON group and HO group,the mRNA and protein expressions of CKAP2 in the HG group were up-regulated;②After siCKAP2 transfection,the cell proliferation ability,migration ability and angiogenesis ability were significantly reduced;③Blocking CKAP2 and blocking VEGF have similar effects on reducing cell abilities in proliferation,migration,and tubular formation.Conclusions CKAP2 may be involved in the proliferation,migration and tube formation of retinal microvascular endothelial cells,and its effect may be similar to VEGF.

关 键 词：糖尿病视网膜病变 细胞骨架蛋白2 视网膜微血管内皮细胞 血管内皮生长因子 

分 类 号：R774.1[医药卫生—眼科]