过氧化物歧化酶和氨基胍对TNF-α诱导的血管内皮细胞凋亡的影响  

Effect of superoxide dismutase and aminoguamidine in tumor necrosis factor-α induced apoptosis on cultured human umbilical vein endothelial cells

作　　者：刘世明[1] 李昭骥[1] 丁月霞[1] 

机构地区：[1]广州医学院附属第二医院心血管内科,广州510260

出　　处：《解剖学研究》2004年第2期88-90,i002,共4页Anatomy Research

基　　金：广东省重点科技攻关项目(2KM04704S)

摘　　要：目的 研究抗氧化剂过氧化物歧化酶(SOD)、诱导型一氧化氮合酶抑制剂氨基胍(AG)对肿瘤坏死因子α(TNF-α)诱导的人脐静脉内皮细胞(ECV-304)凋亡影响及可能的机制。方法 将体外培养的ECV-304的细胞分四组:①空白对照组;②TNF-α组:ECV-304加TNF-α(750、1250、2500、5000、10000 U/ml);③SOD组:ECV-304加TNF-α(2500 U/ml)加SOD(200、400、800 U/ml);④AC组:ECV-304+TNF-α(2500 U/ml)+AC(2、4、8 mmol/L)。采用四唑蓝比色法检测细胞活性;活细胞荧光染色观测细胞凋亡及形态学变化、流式细胞术分析细胞凋亡率;荧光分光光度法测定一氧化氮(NO)。结果 随着TNF-α浓度的增加,ECV-304活性有明显的降低(与对照比P<0.01);SOD或AG均可抑制TNF-α引起的ECV-304活性下降,随着浓度的增加,ECV-304活性有明显的递增趋势,与TNF-α组比较差异有显著性(P<0.01,P<0.05),同时减少NO生成(与TNF-α组比,P<0.05,P<0.002);应用TNF-α(5000 U/ml)的ECV-304凋亡率为27.4％,应用SOD、AG后,两组ECV-304凋亡率分别为17.0％、12.9％。结论 SOD和AG均有抑制TNF-α诱导的内皮细胞凋亡的作用,其抑制作用与减少NO生成有关。Objective To investigate the effect of superoxide dismutase(SOD) and aminoguamidine(AG) in tumor necrosis factor-α(TNF-α) induced apoptosis on cultured human umbilical vein endothelial cells(ECV-304). Methods The cultured ECV-304 were divided into four groups:control,ECV-304 + TNF-α(750,1250,2500,5000,10000 U/ml), ECV-304 + TNF-α(2500 U/ml) + SOD(200,400,800 U/ ml) and ECV-304 + TNF-α(2500 U/ml) + AG(2,4,8 mmol/L) . Treated with different concentration of drugs. Cell surviving was determined by MTT assay,morphological changes were observed by immunofluorescence staining Hoechst 33 258,cell apoptosis were determined by flow cytometry(FCM) ,NO were measured by Gress assay. Results TNF-α induced apoptosis was increased in a dose-dependent manner after 24h incubation( P < 0.01) . The ECV-304 surviving of TNF-αgroup were significantly lower than that of SOD or AG group( P < 0.05, P < 0.05). Compare with TNF-α group, NO was lower in SOD or AG group( P <0.05, P <0.002). The apoptotic rate of SOD, AG and TNF-α group were 27.4% , 17.0% , 12.9% ,respectively. Conclusions TNF-αinduced apoptosis on cultured human umbilical vein endothelial cells is inhibited by superoxide dismutase and aminoguamidine.

关 键 词：TNF-Α ECV-304 SOD 氨基胍 凋亡率 过氧化物歧化酶 诱导 活细胞 一氧化氮(NO) 体外培养 

分 类 号：R541.6[医药卫生—心血管疾病]