人脐静脉内皮细胞中肿瘤坏死因子-α诱导NOSTRIN基因表达上调的实验研究  

The role of PDTC in the inhibition of NOSTRIN overexpression in TNF-α stimulated human umbilical vein endothelial cells

作　　者：相文佩[1] 温子娜[1] 水丽君[1] 胡廉[1] 陈汉平[2] 

机构地区：[1]华中科技大学同济医学院计划生育研究所生殖医学中心,武汉430030 [2]华中科技大学同济医学院附属同济医院妇产科

出　　处：《现代妇产科进展》2010年第7期514-517,522,共5页Progress in Obstetrics and Gynecology

摘　　要：目的:探讨肿瘤坏死因子-α(TNF-α)对人脐静脉内皮细胞(HUVEC)中内皮型一氧化氮合酶运输介导物(NOSTRIN)基因表达的诱导作用以及二硫代氨基甲酸吡咯烷(PDTC)对此诱导作用的抑制。方法:将培养的内皮细胞分为3组:第1组为无任何处理的HUVEC,第2组为子痫前期(PE)患者血清作用的HUVEC,第3组为正常晚孕妇女血清作用的HUVEC。用逆转录-聚合酶链反应(RT-PCR)检测3组细胞中NOSTRIN mR-NA的表达。酶联免疫吸附实验(ELISA)检测PE患者和正常孕妇血清中TNF-α含量。用1,3,5ng/ml3种浓度的TNF-α分别作用3组细胞6h,以及用浓度3ng/ml的TNF-α作用3组细胞分别达6h、12h、24h,RT-PCR检测NOSTRIN mRNA的表达;将PDTC(5mmol/ml)与TNF-α(3ng/ml)同时作用以及TNF-α(3ng/ml)单独作用3组细胞6h,RT-PCR检测NOSTRIN基因mRNA的表达。结果:3组细胞中都有NOSTRIN表达,第2组表达量显著高于第1组和第3组(P<0.01)。ELISA结果显示,PE患者血清中TNF-α含量显著高于正常晚孕妇女(P<0.01)。TNF-α作用HUVEC细胞后NOSTRIN显著高表达,呈时间和剂量依赖关系(P<0.01)。PDTC和TNF-α同时作用的HUVEC与TNF-α单独作用的HUVEC相比,其NOSTRIN表达显著降低(P<0.01)。结论:TNF-α可上调NOSTRIN在HUVEC中的表达。Objective：To study the role of pyrrolidine dithiocarbamate（PDTC） on the gene expression of endothelial nitric oxide synthase traffic inducer（NOSTRIN） in tumor necrosis factor-α（TNF-α）stimulated human umbilical vein endothelial cells（HUVEC）.Methods：The HUVEC were divided into three goups：group 1：control;group2：pretreatment of HUVEC with serum of patients with preeclampsia（PE）;group 3：pretreatment of HUVEC with serum from normal pregnant women.The HUVEC of three goups were exposed to different concentrations of TNF-α（1,3 and 5ng/ml for 6h;3ng/ml for 6h,12h and 24h respectively） or TNF-α（3ng/ml） co-incubated with PDTC （5mmol/ml）for 6h.The expression of NOSTRIN in HUVENS was detected by RT-PCR,and the TNF-α level of serum in PE and normal pregnant women was measured by enzyme linked immunosorbent assay（ELISA）.Results：The gene expression of NOSTRIN in HUVENS in group one was much higher than that of the other two groups（P0.01）.The serum levels of TNF-α were significantly higher in women with PE than in the normal pregnant women（P0.01）. NOSTRIN expression level was up-regulated by TNF-α in dose-dependent and time-dependent manner in human umbilical vein endothelial cells（P0.01）.The expression of NOSTRIN in the cells treated with TNF-α and PDTC was significantly lower than that treated with TNF-α alone（P0.01）.Conclusion：The present study suggests that TNF-α can up-regulate the gene expression of NOSTRIN in HUVEC.

关 键 词：肿瘤坏死因子-Α 内皮型一氧化氮合酶运输介导物 二硫代氨基甲酸吡咯烷 人脐静脉内皮细胞 

分 类 号：R714.244[医药卫生—妇产科学]