小檗碱调节AKT/NF-κB信号通路对高糖诱导的足细胞损伤、凋亡和迁移的影响  

Effects of berberine on podocyte injury,apoptosis and migration induced by high glucose through regulating AKT/NF-κB signaling pathway

作　　者：汪佳佳 胡亚琴 吴昊 唐丽琴[1,2] 魏伟 Wang Jiajia;Hu Yaqin;Wu Hao(Institute of Clinical Pharmacology,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学临床药理研究所,合肥230032 [2]中国科学技术大学附属第一医院,合肥230001

出　　处：《安徽医科大学学报》2021年第6期845-850,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81773955)。

摘　　要：目的探究小檗碱(BBR)对高糖环境下足细胞的保护作用及其机制。方法取指数生长期的足细胞进行实验,将细胞分为五组:正常组,高糖组,BBR 30、60、90μmol/L组。Annexin FITC/PI双染法检测足细胞凋亡率,高内涵成像系统计算细胞数量,Transwell和细胞划痕实验检测其迁移能力,Western blot检测蛋白激酶B/核转录因子κB(AKT/NF-κB)信号通路相关蛋白、细胞促凋亡蛋白(Bim)和足细胞标志性蛋白WT-1、Podocin、desmin的表达水平。结果与正常组相比,Annexin FITC/PI双染法结果显示,高糖组足细胞凋亡率上升(P<0.01);高内涵细胞成像系统显示足细胞数量减少(P<0.01);Transwell结果显示迁移至下室的足细胞数增加(P<0.01);细胞划痕实验结果显示划痕愈合率上升(P<0.01);Western blot结果显示,高糖组足细胞中p-AKT、p-p65、p-IκBα、Bim、desmin蛋白表达水平升高(P<0.01),WT-1、Podocin蛋白表达水平降低(P<0.01)。与高糖组相比,BBR给药后足细胞凋亡率下降(P<0.05);不同浓度BBR给药组的足细胞数量增多(P<0.01);迁移至下室的足细胞数有所减少(P<0.05);划痕愈合率下降(P<0.01);另外,BBR给药降低了p-AKT、p-p65、p-IκBα、Bim、desmin蛋白表达水平(P<0.05),同时升高了WT-1、Podocin蛋白表达水平(P<0.01)。结论BBR可能通过AKT/NF-κB信号通路改善高糖诱导的足细胞损伤、抑制足细胞凋亡、减弱足细胞迁移能力。Objective To study the protective effect of berberine(BBR)on podocytes in high glucose environment and to discuss its mechanism.Methods The exponential growth stage of the glomerular podocyte was selected.The podocyte were divided into 5 groups,including normal control group,high-glucose model group and BBR was given in concentrations of 30,60 and 90μmol/L.Annexin FITC/PI double staining was used to detect the apoptosis rate of podocytes.High-intension imaging system was used to detect cell proliferation,podocyte migration was measured by transwell and cell scratch assay,Western blot detected the expression of AKT/NF-κB signaling pathway related proteins,apoptotic protein Bim,and the podocyte marker proteins WT-1,Podocin and desmin.Results Compared with the normal group,the results of Annexin FITC/PI double staining showed that the apoptosis rate of podocytes in the high glucose group increased(P<0.01);The high-content imaging system showed that the number of podocytes in the high glucose group decreased(P<0.01);Transwell showed that the number of podocytes migrate ability increased in the high glucose group(P<0.01);The cell scratch showed that the scratch healing rate of the high glucose group increased(P<0.01);Western blot showed that the expression levels of p-AKT,p-p65,p-IκBα,Bim,and desmin protein in the podocytes of the high glucose increased(P<0.01),and the expression of WT-1 and Podocin proteins decreased(P<0.01).Compared with the high glucose group,the apoptosis rate of podocytes decreased after add BBR(P<0.05);the number of podocytes increased in the different concentrations of BBR(P<0.01);the number of podocyte migration was lower(P<0.05);Scratch healing rate decreased(P<0.01);In addition,BBR reduced the protein expression levels of p-AKT,p-p65,p-IκBα,Bim and desmin(P<0.05),the expression level of WT-1 and Podocin protein increased(P<0.01).Conclusion BBR may improve the podocyte injury,inhibit the podocyte apoptosis,and weaken the podocyte migration ability induced by high glucose through regulat

关 键 词：小檗碱 糖尿病肾病 足细胞 蛋白激酶B 核因子ΚB 信号通路 

分 类 号：R285.5[医药卫生—中药学]