Klotho RNA干扰对人主动脉血管平滑肌细胞增殖、迁移及氧化应激的影响  

Effects of Klotho RNA Interference on Proliferation,Migration and Oxidative Stress in Human Aortic Vascular Smooth Muscle Cells

作　　者：魏建行 梁荃 黄仕琼 李利华 Wei Jianhang;Liang Quan;Huang Shiqiong;Li Lihua(Department of Gerontology,The First Affiliated Hospital of Dali University,Dali,Yunnan 671000,China)

机构地区：[1]大理大学第一附属医院老年病科,云南大理671000

出　　处：《大理大学学报》2021年第4期30-35,共6页Journal of Dali University

基　　金：国家自然科学基金项目(81460084;81660072;81860084);云南省中青年学术和技术带头人培养项目(2015HB056);云南省医学学科带头人培养项目(D-201672);大理大学高血压防治创新团队项目(ZKPY2019304);云南省万人计划青年拔尖人才项目(云人社发[2018]73号)。

摘　　要：目的:探讨Klotho RNA干扰对人主动脉血管平滑肌细胞(HA-VSMC)增殖、迁移及氧化应激的影响。方法:采用慢病毒抑制HA-VSMC的Klotho表达,通过荧光显微镜、荧光定量聚合酶链反应及蛋白质印迹法验证转染效率。采用CCK-8法检测细胞的增殖,Transwell法检测细胞的迁移,微量法检测细胞内超氧化物歧化酶(SOD)以及丙二醛(MDA)的含量,采用荧光定量聚合酶链反应检测NOX1、NOX2、NOX3、NOX5以及DUOX2相对表达量。结果:慢病毒转染HA-VSMC后Klotho表达明显下调。与空白对照孔相比,Klotho表达减少能显著增加HA-VSMC增殖、迁移能力,烟酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX1,NOX2,NOX3,NOX5和DUOX2)mRNA表达水平显著增加,细胞裂解物中MDA明显增加以及SOD水平显著降低。结论:Klotho表达减少可促进HA-VSMC增殖、迁移以及氧化应激。Objective:To examine the effects of Klotho RNA interference on proliferation,migration and oxidative stress in human aortic vascular smooth muscle cells(HA-VSMC).Methods:Lentivirus was used to inhibit the expression of Klotho in HA-VSMC,and the transfection efficiency was verified by fluorescence microscopy,fluorescence quantitative polymerase chain reaction and the Western blot.CCK-8 method was used to detect cell proliferation,and Transwell method was used to detect cell migration.The intracellular superoxide dismutase(SOD)and malondialdehyde(MDA)content was detected by the micro method.The relative expressions of NOX1,NOX2,NOX3,NOX5 and DUOX2 were detected by fluorescence quantitative polymerase chain reaction.Results:The expression of Klotho in HA-VSMC was significantly down-regulated after lentivirus transfection.Compared with the blank control well,the decrease of Klotho expression could significantly increase the proliferation and migration of HA-VSMC,and nicotinamide adenine dinucleotide phosphate oxidase(NOX1,NOX2,NOX3,NOX5 and DUOX2)mRNA expression were increased significantly.Meanwhile,MDA levels increased significantly,while the SOD activity levels decreased significantly in cell lysates after transfection.Conclusion:Klotho deficiency could promote HA-VSMC proliferation,migration and oxidative stress.

关 键 词：KLOTHO 主动脉血管平滑肌细胞 增殖 迁移 氧化应激 

分 类 号：R543[医药卫生—心血管疾病]