解放军医学杂志2017，Vol.42Issue(3)：190-195,6.DOI:10.11855/j.issn.0577-7402.2017.03.03
ROBO4过表达抑制高糖条件下人肾小球内皮细胞高通透性的体外研究
Over-expression of ROBO4 reduces permeability of human renal glomerular endothelial cells in high glucose medium
摘要
Abstract
Objective To investigate the effect of over-expression of ROBO4 on permeability of human renal glomerular endothelial cells (HRGECs) in high glucose medium.Methods HRGECs infected with recombinant lentiviral vector ROBO4 were cultured in high glucose or low glucose medium in vitro.The protein levels of ROBO4 and ARF6 in each group were detected by Western blotting.The endothelial permeability was measured by the effiux of fluorescein isothiocyanate-dextran (FITC-Dextran)permeated through the monolayer endothelial cells using Transwell cell model system.The cell viability after lentivirus transfection was measured by CCK8 assay.Results The transfection rate of lentiviruses in HRGECs reached 80％ 72h after,and obvious overexpression of ROBO4 protein was in transformed cells compared with the empty vector group (P＜0.05).The lentivirus-mediated ROBO4 transfection did not affect cell viability of HRGECs.Compared with the low glucose group,the expression of ROBO4 increased obviously after 12h,but declined after 24h (P＜0.05),and reached to minimun after 72h (P＜0.05).On the contrary,the expression of ARF6 increased after 12h,and the increase reached to the maximum after 72h (P＜0.05).Furthermore,the vascular permeability increased gradually after 24h,and reached to the maximum after 72h (P＜0.05) in high glucose group.Compared with the empty vector group,the over-expression of ROBO4 inhibited the expression of ARF6 significantly,and the FITC-Dextran permeability reduced obviously.Conclusion Over-expression of ROBO4 may significantly enhance the barrier functions of HRGEC in high glucose medium,and ROBO4 activation may be a potential therapeutic approach in diabetic nephropathy.关键词
ROBO4/ARF6/通透性/基因过表达/糖尿病肾病Key words
ROBO4/ARF6/permeability/gene over-expression/diabetic nephropathies分类
医药卫生引用本文复制引用
唐璐瑶,刘军辉,候世会,官涛,黄云剑..ROBO4过表达抑制高糖条件下人肾小球内皮细胞高通透性的体外研究[J].解放军医学杂志,2017,42(3):190-195,6.基金项目
国家自然科学基金(81370821) （81370821）
重庆市自然科学基金重点项目(cstc2013jjB10023)This work was supported by the National Natural Science Foundation of China (81370821),and the Natural Science Foundation of Chongqing City (cstc2013jjB10023) （cstc2013jjB10023）
