驱动蛋白Kif5b对人脐静脉内皮细胞血管形成的作用  

Effects study of a kinesin Kif5b on the angiogenesis of HUVECs

作　　者：罗语思 肖美兰 陈妍 张科 LUO Yusi;XIAO Meilan;CHEN Yan;ZHANG Ke(Department of Emergency,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Key Laboratory of Optimal Utilization of Natural Medicine Resources,School of Pharmaceutical Sciences,Guizhou Medical University,Guiyang 550025,Guizhou,China;Key and Characteristic Laboratory of Modern Pathogen Biology&Parasitology Division,Basic Medical College&Key and Characteristic Laboratory of the High Efficacy Application of Natural Medicinal Resources,School of Pharmaceutical Sciences,Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学附属医院急诊科,贵州贵阳550004 [2]贵州医科大学药学院贵州省高等学校天然药物药理与成药性评价重点实验室,贵州贵阳550025 [3]贵州医科大学基础医学院现代病原生物学特色重点实验室&基础医学院寄生虫学教研室&药学院天然药物资源优效利用重点实验室,贵州贵阳550025

出　　处：《贵州医科大学学报》2021年第1期32-37,共6页Journal of Guizhou Medical University

基　　金：贵州省卫健委科学技术基金项目(gzwjkj2018-1-022);贵州省科技创新团队项目[黔科合人才团队(2015)4025];贵州省科技厅省校合作联合基金项目[黔科合LH字(2016)7245];贵州省高层次创新型人才百层次人才项目[贵州省科技厅黔科合人才(2015)4029];贵州省教育厅自然科学基金优秀人才计划[黔教合KY字(2018)048]。

摘　　要：目的 探讨驱动蛋白Kif5b对人脐静脉内皮细胞(HUVECs)血管形成的作用。方法 以人Kif5b基因序列为模板设计小发夹RNA(sh RNA)序列,同时设阴性对照sh RNA,均以p LL3.7慢病毒包装系统包装sh RNA,随后感染HUVECs敲减Kif5b蛋白,最后将Kif5b敲减的HUVECs作为Kif5b实验组,阴性对照组作为Kif5b对照组;采用Western blot实验检测2组HUVECs中Kif5b的表达,采用细胞成管实验和细胞划痕实验分别检测2组HUVECs的细胞成管率和细胞迁移速度。结果 Kif5b实验组HUVECs的Kif5b表达较Kif5b对照组下降,差异有统计学意义(P<0.05);Kif5b实验组HUVECs细胞成管率明显低于Kif5b对照组,差异有高度统计学意义(P<0.000 1);Kif5b实验组HUVECs细胞迁移速度明显低于Kif5b对照组,差异有高度统计学意义(P<0.000 1)。结论 在体外实验中,驱动蛋白Kif5b能够正向调控HUVECs血管形成。Objective To investigate the effect of a kinesin,Kif5 b on the angiogenesis of human umbilical vein endothelial cells(HUVECs).Methods The small hairpin RNA(shRNA) was designed with human Kif5 b gene sequence as template,and negative control sh RNA was also designed.shRNAs were packaged with pLL3.7 lentivirus packaging system,then infect the HUVECs cells to knock down its Kif5 b protein.And then set HUVECs with Kif5 b knocked down as Kif5 b test group,the negative control group as Kif5 b control group.The Kif5 b expression level of two groups was measured by western blot.And the rate of tube formation and cell migration of two groups was tested by the tube formation assay and scratch assay.Results Compared with control group,immunoblotting results shows that the Kif5 b expression level in p LL3.7-shKif5 b HUVECs decreased significantly.And the results of tube formation assay and scratch assay demonstrated the ability of tube formation and migration of p LL3.7-shKif5 b HUVECs were impaired significantly.Conclusion The kinesin,Kif5 b can promote the angiogenesis of HUVECs in vitro.

关 键 词：驱动蛋白 Kif5b蛋白 人脐静脉内皮细胞 小发夹RNA 细胞成管率 细胞迁移速度 血管形成 

分 类 号：R730.2[医药卫生—肿瘤]