干扰Sestrin2表达对ox-LDL诱导的人脐静脉内皮细胞凋亡的影响  

Effects of Sestrin2 Inhibition on Human Umbilical Vein Endothelial Cells Apoptosis Induced by ox-LDL

作　　者：刘锋[1,2] 胡红娟[2] 范文静[3] 

机构地区：[1]南华大学研究生院 [2]南华大学护理学院,湖南省衡阳市421001 [3]南华大学附属第二医院

出　　处：《中国动脉硬化杂志》2016年第4期350-354,共5页Chinese Journal of Arteriosclerosis

基　　金：湖南省自然科学基金项目(2015JJ2118);湖南省教育厅重点项目(15A166);湖南省卫计委科研项目(20160233);衡阳市科技局项目(2015KJ16);南华大学研究生科技创新项目(2015XCX38)

摘　　要：目的观察干扰Sestrin2表达对氧化型低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVEC)凋亡的影响。方法用ox-LDL处理HUVEC复制内皮细胞损伤模型。Western blot检测不同浓度ox-LDL处理HUVEC不同时间Sestrin2的蛋白表达水平及转染Sestrin2 siRNA后Sestrin2的蛋白表达水平;流式细胞术检测干扰Sestrin2表达后对ox-LDL诱导的HUVEC凋亡率的影响;Western blot检测干扰Sestrin2表达后对ox-LDL诱导的HUVEC中Caspase-3表达的影响及ox-LDL对HUVEC中JNK通路的影响。结果不同浓度(20、50和100 mg/L)的ox-LDL处理HUVEC 24 h均能明显上调Sestrin2表达;50 mg/L ox-LDL处理HUVEC不同时间(24 h和48 h)均能明显上调Sestrin2表达,24 h达高峰;转染Sestrin2 siRNA能明显抑制Sestrin2的表达;ox-LDL能明显诱导HUVEC凋亡,转染Sestrin2 siRNA能明显促进由ox-LDL诱导的HUVEC凋亡;ox-LDL能明显诱导p-JNK和p-c-Jun的表达,且SP600125能明显抑制由ox-LDL诱导的Sestrin2表达。结论 ox-LDL通过JNK/c-Jun信号通路诱导Sestrin2表达,干扰Sestrin2表达能明显促进ox-LDL诱导的HUVEC凋亡。Aim To investigate the effect of Sestrin2 inhibition on apoptosis of human umbilical vein endothelial cells induced by oxidized low density lipoprotein（ ox-LDL）.Methods The cell injury model was induced by oxLDL.Western blot was used to detect protein expression of Sestrin2 in human umbilical vein endothelial cells treated with different concentrations of ox-LDL at different time.Protein expression of Sestrin2 was observed by Western blot after transfection with Sestrin2 siRNA.Human umbilical vein endothelial cells were transfected with Sestrin2 siRNA,then treated with ox-LDL,cell apoptosis was detected by flow cytometry,Caspase-3 expression and JNK pathway were observed by Western blot.Results Sestrin2 expression was significantly induced after treatment at different concentrations（ 20,50 and 100 mg/L） of ox-LDL for 24 h.Moreover,human umbilical vein endothelial cells were incubated with50 mg /L ox-LDL for various periods of time ranging from 0 to 48 h.The results showed a significant increase of Sestrin2 at protein levels in 24 h and 48 h,and it was the highest level in 24 h.Transfection with Sestrin2 siRNA significantly inhibited Sestrin2 expresssion.ox-LDL significantly induced cell apoptosis,transfection with Sestrin2 siRNA increased the effect.Expression of p-JNK and p-c-Jun was induced by ox-LDL,and this effect could be inhibited by SP600125.Conclusions Ox-LDL induces Sestrin2 expression through JNK /c-Jun pathway,inhibition of Sestrin2 expression significantly increases human umbilical vein endothelial cells apoptosis induced by ox-LDL.

关 键 词：Sestrin2 氧化型低密度脂蛋白 人脐静脉内皮细胞 细胞凋亡 

分 类 号：R363[医药卫生—病理学]