黄芪甲苷保护嘌呤霉素氨基核苷诱导的足细胞损伤及机制探讨  

The protective effect of astragaloside IV on puromycin aminonucleoside induced podocyte injury and its potential mechanism

作　　者：龚建光[1] 詹慧芳 金娟[1] 赵黎[1] 李一文[1] 何强[1] GONG Jian-guang;ZHAN Hui-fang;JIN Juan;ZHAO Li;LI Yi-wen;HE Qiang(Department of Nephrology,Zhejiang Province People’s Hospital/People’s Hospital of Hangzhou Medical College,Hangzhou 310014,China)

机构地区：[1]浙江省人民医院/杭州医学院附属人民医院肾脏病科,杭州310014 [2]浙江大学校医院紫金港校区,杭州310058

出　　处：《临床肾脏病杂志》2020年第4期303-309,共7页Journal Of Clinical Nephrology

基　　金：浙江省中医药管理局项目(No.2017ZA008);浙江省自然基金项目(No.LZ17H050001)。

摘　　要：目的探讨黄芪甲苷对嘌呤霉素氨基核苷诱导的足细胞凋亡的影响及可能机制。方法构建嘌呤霉素氨基核苷诱导的足细胞损伤模型,根据不同处理将足细胞分成:Control组、PAN组(50μg/mL嘌呤霉素氨基核苷处理24 h)、AS-IV组(分别用5μg/mL、15μg/mL、30μg/mL黄芪甲苷处理24 h)、AS-IV+PAN组(先用黄芪甲苷预处理30 min,再用50μg/mL嘌呤霉素氨基核苷处理24 h)。采用CCK-8检测足细胞生长活力,Annexin V/PI双染法检测细胞凋亡,免疫荧光法检测足细胞标记蛋白,Western blot检测caspase-3、cleaved caspase-3及VEGFR2/GIV通路的蛋白水平。结果与Control组比较,PAN组足细胞的凋亡率显著增加(P<0.05),nephrin、podocin、p-AKT的蛋白水平明显降低(P<0.05),cleaved caspase-3、p-VEGFR2、p-GIV的蛋白水平均明显上调(P<0.05);与PAN组比较,AS-IV+PAN组足细胞的凋亡率显著降低(P<0.05),nephrin、podocin的蛋白水平明显增加(P<0.05),cleaved caspase-3的蛋白水平显著下降(P<0.05),p-VEGFR2、p-GIV、p-AKT的蛋白水平进一步上调(P<0.05)。结论黄芪甲苷可以保护嘌呤霉素氨基核苷诱导的足细胞损伤,VEGFR2/GIV通路可能参与其调节的作用机制。Objective To explore the inhibitory effect of astragaloside IV(AS-IV)on podocyte injury induced by puromycin aminonucleoside(PAN)and its underlying mechanism.Methods A PAN-induced podocyte injury model was established to evaluate the effect of AS-IV on podocyte apoptosis.Based on different treatment,The MPC5 podocytes were randomly divided into 4 groups:control group,PAN group(50μg/mL PAN,24h),AS-IV group(5μg/mL,15μg/mL,30μg/mL AS-IV,24 h)and AS-IV+PAN group(pretreated with AS-IV for 30 min,then treated with PAN for 24 h).CCK-8 was used to detect growth activity of podocytes,Annexin V/PI double staining method to detect podocyte apoptosis,immunofluorescence assay to detect the marker protein in podocytes,Western blot to detect the protein expression levels of Caspase-3,cleaved Caspase-3 and VEGFR2/GIV pathway.Results Compared with the control group,in the PAN group the podocyte apoptotic rate was significantly increased(P<0.05),the expression of nephrin and podocin was decreased notably(P<0.05),and the levels of cleaved caspase-3,p-VEGFR2 and p-GIV were up-regulated significantly(P<0.05).Compared with the PAN group,the podocyte apoptotic rate and protein levels of cleaved caspase-3 was dramatically declined in the AS-IV+PAN group(P<0.05),accompanied with obvious increase of nephrin and podocin levels(P<0.05),significant decrease of cleaved caspase-3,and further upregulation of p-VEGFR,p-GIV and p-AKT(P<0.05).Conclusions Astragaloside IV attenuates puromycin aminonucleoside induced podocyte injury and it may be mediated by the VEGFR2/GIV signaling pathway.

关 键 词：黄芪甲苷 嘌呤霉素氨基核苷 足细胞凋亡 VEGFR2/GIV通路 

分 类 号：R285.5[医药卫生—中药学]