二苯乙烯苷对过氧化氢诱导内皮细胞凋亡及XIAP和p53表达的影响  

Effects of TSG on H_2O_2-induced Apoptosis and Expressions of XIAP and p53 in HUVECs

作　　者：杨滢[1] 高细强[2] 龙石银[3] 

机构地区：[1]蚌埠医学院生物化学与分子生物学教研室,蚌埠233030 [2]湖南医药学院附属怀化市第二人民医院,怀化418200 [3]南华大学生物化学与分子生物学教研室,衡阳421001

出　　处：《华中科技大学学报（医学版）》2015年第6期629-633,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30800474);湖南省卫生计生委资助项目(No.B2015-49);湖南省高层次卫生人才"225"工程项目基金;湖南省科技厅资助项目(No.2015SK2038);南华大学十二五科技创新团队基金;南华大学留学归国人员启动基金资助

摘　　要：目的探讨二苯乙烯苷(TSG)对过氧化氢(H_2O_2)诱导的人脐静脉内皮细胞凋亡及对X连锁凋亡抑制蛋白(XIAP)、p53基因表达的影响。方法体外培养人脐静脉内皮细胞,并分为:对照组、300μmol/L H_2O_2组、1μmol/L TSG+300μmol/L H_2O_2组、10μmol/L TSG+300μmol/L H_2O_2组、100μmol/L TSG+300μmol/L H_2O_2组及30μmol/L Embelin+10μmol/L TSG+300μmol/L H_2O_2组。采用Hoechst 33258染色观察凋亡细胞形态,RT-PCR检测XIAP、p53、Caspase-3mRNA表达,Western blot检测XIAP、p53、Caspase-3蛋白的表达。结果与正常对照组相比,H_2O_2处理组凋亡细胞数增多,XIAP mRNA及蛋白表达水平显著降低,p53表达明显升高;与H_2O_2组相比,10、100μmol/L TSG预处理24h后,细胞凋亡减少,XIAP表达升高,p53的表达下降;与10μmol/L TSG预处理组相比,30μmol/L Embelin预处理组凋亡细胞数量增加,Caspase-3表达升高。结论 TSG能促进XIAP的表达,抑制p53表达,拮抗H_2O_2诱导的人脐静脉内皮细胞损伤。Objective To study the effects of tetrahydroxy stilbene glucoside（TSG）on H_2O_2-induced apoptosis of human umbilical vein endothelial cells（HUVECs）and on the expressions of X-linked inhibitor of apoptosis protein（XIAP）and p53.Methods HUVECs were cultured in vitro and divided into 6groups：control group,300μmol/L H_2O_2 group,1μmol/L TSG＋ 300μmol/L H_2O_2 group,10μmol/L TSG＋300μmol/L H_2O_2 group,100μmol/L TSG＋ 300μmol/L H_2O_2 group,30μmol/L Embelin＋10μmol/L TSG＋300μmol/L H_2O_2 group.The morphology of apoptotic cells was observed by Hoechst33258 staining.The mRNA and protein expressions of XIAP,p53,Caspase-3were detected by RT-PCR and Western blotting,respectively.Results The number of apoptotic cells and the expression level of p53 were significantly increased while the expression level of XIAP was dramatically decreased in H_2O_2 group as compared with control group.The expression level of p53 and the number of apoptotic cells were down-regulated while the expression level of XIAP was up-regulated after treatment with10 or 100μmol/L TSG when compared with H_2O_2 group.Moreover,compared with those in 10μmol/L TSG group,the number of apoptotic cells and the expression of Caspase-3were significantly enhanced after pretreatment with 30μmol/L Embelin for 6h.Conclusion TSG can inhibit H_2O_2-induced apoptosis of HUVECs by down-regulating the expression level of p53 and up-regulating the expression level of XIAP.

关 键 词：动脉粥样硬化 二苯乙烯苷 X连锁凋亡抑制蛋白 P53 

分 类 号：R543.5[医药卫生—心血管疾病]