过表达sFRP2基因对脂多糖诱导人脐静脉内皮细胞凋亡的影响及其分子机制  

Effect of overexpression of sFPR2 gene on apoptosis of human umbilical vein endothelial cells induced by lipopolysaccharide and its molecular mechanism

作　　者：蒙艳 白漪 霍松 MENG Yan;BAI Yi;HUO Song(Honghe Hospital Affiliated to Kunming Medical University,Honghe 661000,China)

机构地区：[1]昆明医科大学附属红河医院,云南红河661000

出　　处：《山东医药》2021年第11期20-23,共4页Shandong Medical Journal

基　　金：云南省基础研究计划项目(2017FB226)。

摘　　要：目的观察过表达分泌型卷曲相关蛋白2(sFRP2)对脂多糖(LPS)诱导人脐静脉内皮细胞(HUVECs)凋亡的影响,并探讨其可能的作用机制。方法将HUVECs分为两部分,一部分加入不同浓度LPS(0、5、10、15、20μg/mL),另一部分作为对照细胞不予特殊处理,作用12 h;采用CCK-8法检测其细胞存活指数,结果显示加入10、15、20μg/mL LPS作用12 h的HUVECs细胞存活指数明显低于对照细胞(P均<0.01),选择10μg/mL LPS作用12 h作为后续实验条件。将HUVECs分为过表达组和阴性对照组,分别转染sFRP2过表达质粒及阴性对照慢病毒质粒,转染48 h后加入10μg/mL LPS作用12 h;采用实时荧光定量PCR法和Western boltting法检测sFRP2 mRNA和蛋白表达,流式细胞术检测细胞凋亡率,Western boltting法检测凋亡相关蛋白Bax、Cleaved Caspase-3、Bcl-2和Wnt/β-catenin信号通路相关蛋白Wnt1、β-catenin、Cyclin D1表达。结果过表达组与阴性对照组sFRP2 mRNA相对表达量分别为2.69±0.13、1.04±0.21,蛋白相对表达量分别为3.89±0.18、1.05±0.24,细胞凋亡率分别为13.32%±1.98%、20.90%±1.72%,两组比较P均<0.01。过表达组Cleaved Caspase-3、Bax及Wnt1、β-catenin、Cyclin D1表达均低于阴性对照组,Bcl-2表达高于阴性对照组(P均<0.05)。结论过表达sFRP2可减少LPS诱导的HUVECs细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关。Objective To observe the effect of secreted frizzled-related protein 2(sFRP2)overexpression on the apoptosis of human umbilical vein endothelial cells(HUVECs)induced by lipopolysaccharide(LPS),and to explore the possible mechanism.Methods HUVECs were divided into two parts.One part was added with different concentrations of LPS(0,5,10,15 and 20μg/mL),and the other part was treated as control cells without special treatment and cultured for 12 h.CCK-8 was used to detect the cell survival index,the results showed that the survival index of HUVECs treated with 10,15 and 20μg/mL LPS for 12 h was significantly lower than that of the control cells(all P<0.01),and 10μg/mL LPS for 12 h was selected as the subsequent experimental conditions.HUVECs were divided into the overexpression group and negative control group,respectively,transfected with SFRP2 overexpression and negative control lentivirus plasmid;at 48 h after transfection,10μg/mL LPS was added for 12 h.The expression of SFRP2 mRNA and protein was detected by realtime fluorescence quantitative PCR and Western boltting.Apoptosis rate was detected by flow cytometry.The expression levels of apoptosis-related proteins Bax,Cleaved caspase-3,Bcl-2 and Wnt/β-catenin signaling pathway related proteins Wnt1,β-catenin and Cyclin D1 were detected by Western boltting.Results The relative expression levels of SFRP2 mRNA in the overexpression group and negative control group were 1.04±0.21 and 2.69±0.13,respectively,and the relative expression levels of protein were 3.89±0.18 and 1.05±0.24.The apoptosis rates of the overexpression group and the negative control group were 13.32%±1.98%and 20.90%±1.72%,and there were significant difference be⁃tween the two groups(all P<0.01).The expression of Cleaved caspase-3,Bax,Wnt1,β-catenin and Cyclin D1 decreased in the overexpression group,and the expression of Bcl-2 increased(all P<0.05).Conclusion Overexpression of sFRP2 can reduce the apoptosis of HUVECs induced by LPS,and the mechanism may be related to the inhibition o

关 键 词：内皮细胞 SFRP2 脂多糖 细胞凋亡 WNT/Β-CATENIN信号通路 

分 类 号：R285.5[医药卫生—中药学]