VEGFR1下调对Aβ1-42诱导人脑微血管内皮细胞衰老及PI3K信号通路的影响  

Effects of down-regulation of VEGFR1 on Aβ1-42-induced aging of human microvascular endothelial cells and PI3K signaling pathway in vitro

作　　者：刘洁 LIU Jie(Department of Genetic Teaching and Research,Xi’an Medical College,Shaanxi,Xi’an 710002,China)

机构地区：[1]西安医学院基础医学部细胞与遗传教研室,西安市710002

出　　处：《河北医药》2021年第7期990-993,999,共5页Hebei Medical Journal

基　　金：国家自然科学基金资助项目(编号:81471415);陕西省教育厅专项科研计划项目(编号:17JK0666);西安医学院2016年度教育教学改革研究项目(编号:2016JG-39)。

摘　　要：目的探讨小干扰RNA(siRNA)沉默血管内皮细胞生长因子受体1(VEGFR1)基因表达对β淀粉样蛋白1-42(Aβ1-42)诱导人脑微血管内皮细胞HBMEC衰老及磷酸肌醇3激酶(PI3K)信号通路的影响。方法体外培养人脑微血管内皮细胞HBMEC,将细胞分为4组:HBMEC组(不转染,不采用Aβ1-42诱导),HBMEC+Aβ1-42组(不转染,50μmol/L Aβ1-42诱导)、neg+Aβ1-42组(转染VEGFR1siRNA-neg,50μmol/L Aβ1-42诱导),VEGFR1 siRNA+Aβ1-42组(转染VEGFR1siRNA,50μmol/L Aβ1-42诱导)。转染48 h后,利用人胆囊收缩素/缩胆囊素八肽(CCK-8)试剂盒检测细胞活力,用β-半乳糖苷酶染色试剂盒(B-gal Kit)检测细胞衰老情况,Hochest 33342/PI双染法观察细胞凋亡情况,蛋白印迹(WB)法检测细胞VEGFR1、p-Akt、p-mTOR、PI3K蛋白表达。结果与HBMEC组比较,HBMEC+Aβ1-42组、neg+Aβ1-42组、VEGFR1 siRNA+Aβ1-42组细胞存活率及细胞中VEGFR1、p-Akt、p-mTOR、PI3K蛋白表达水平均显著下降(P<0.05),衰老及凋亡细胞均增多(P<0.05);与HBMEC+Aβ1-42组、neg+Aβ1-42组比较,VEGFR1 siRNA+Aβ1-42组细胞存活率及细胞中VEGFR1、p-Akt、p-mTOR、PI3K蛋白表达水平均显著下降(P<0.05)、衰老及凋亡细胞均增多(P<0.05)。结论沉默VEGFR1可能通过抑制PI3K信号通路活化,促进Aβ1-42诱导的人脑微血管内皮细胞衰老及凋亡。Objective To investigate the effects of small interfering RNA(siRNA)silencing VEGFR1 gene expression on amyloidβ1-42(Aβ1-42)-induced HBMEC aging and phosphoinositide 3 kinase(PI3K)signaling pathway in human microvascular endothelial cells(HBMEC)in vitro.Methods The HBMECs were cultured in vitro and divided into four groups:HBMEC group(no transfection,no Aβ1-42 induction),HBMEC+Aβ1-42 group(no transfection,50μmol/L Aβ1-42 induction),neg+Aβ1-42 group(transfected with VEGFR1siRNA-neg 50μmol/L Aβ1-42 induction),vegfr1sirna+Aβ1-42 group(transfected with VEGFR1 siRNA,50μmol/L Aβ1-42 induction).At 48 hours after transfection,the cell viability was detected by human cholecystokinin/cholecystokinin octapeptide(CCK-8)kit,andβ-galactosidase(β-gal)kit was used to detect the cell aging,and the cell apoptosis was observed by Hochst 33342/PI double staining,moreover,the expression levels of VEGFR1,p-Akt,p-mTOR and PI3K proteins were detected by Western Blot.Results As compared with those in HBMEC group,the cell survival rate and expression levels of VEGFR1,p-Akt,p-mTOR and PI3K protein in HBMEC+Aβ1-42 group,neg+Aβ1-42 group,VEGFR1 siRNA+Aβ1-42 group were decreased significantly(P<0.05),and he aging and apoptotic cells were increased significantly(P<0.05).As compared with those in HBMEC+Aβ1-42 group and neg+Aβ1-42 group,the cell survival rate and expression levels of VEGFR1,p-Akt,p-mTOR and PI3K protein in HBMEC+Aβ1-42 group,neg+Aβ1-42 group,VEGFR1 siRNA+Aβ1-42 group were decreased significantly(P<0.05),and the aging and apoptotic cells were increased significantly(P<0.05).Conclusion Silencing VEGFR1 may promote the senescence and apoptosis of human microvascular endothelial cells induced by aβ1-42 through inhibiting the activation of PI3K signaling pathway.

关 键 词：β淀粉样蛋白1-42 人脑微血管内皮细胞 衰老 血管内皮细胞生长因子受体 磷酸肌醇3激酶通路 

分 类 号：R741[医药卫生—神经病学与精神病学]