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基因(eNOS)表达的作用以及PDTC对此作用的改变。方法将培养的内皮细胞分为3组:1组为无任何处理的HUVEC,2组为子痫前期(PE)患者血清作用的HUVEC,3组为用正常晚孕妇女血清作用的HUVEC。用逆转录-聚合酶链反应检测三组细胞中eNOS mRNA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1ng/ml、3ng/ml、5ng/ml 3种浓度的TNF-α分别作用三组细胞6h,以及用浓度为3ng/ml的TNF-α作用三组细胞分别长达6h、12h、24h,逆转录-聚合酶链反应(RT-PCR)检测eNOS mRNA的表达;将二硫代氨基甲酸吡硌烷(PDTC,5mmol/ml)与TNF-α(3ng/ml)同时作用以及TNF-α(3ng/ml)单独作用三组细胞6h,RT-PCR检测eNOS基因mRNA的表达。结果三组细胞中都有eNOS表达,2组表达量显著低于1组和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eNOS表达显著低表达,并成时间和剂量依赖关系(P<0.01)。PDTC和TNF-α同时作用的HUVEC和TNF-α单独作用的HUVEC相比,其eNOS表达显著增高(P<0.01)。结论可下调eNOS在HUVEC中的表达,PDTC可改变TNF-α诱导的eNOSN表达的下调。
Objective：To study the role of pyrrolidine dithiocarbamate（PDTC） in the inhibition of endothelial nitric oxide synthase（eNOS） downexpression by tumor necrosis factor-α（TNF-α） in human umbilical vein endothelial cells（HUVEC）.Methods：The HUVEC were divided into three groups： Group 1： HUVEC no any factors treated with,group 2： HUVEC were treated with serum from the women with pre-eclampsia（PE）,group 3： HUVEC were treated with serum from the normal pregnant women.RT-PCR was applied to detect eNOS expression;the TNF-α level of serum was measured by Enzyme-linked immunosorbent assay（ELISA） in normal pregnant women and women with PE.Three groups HUVEC were treated with TNF-α（1ng/ml,3ng/ml,5ngU/ml） for 6h respectively,and treated with TNF-α（3ng/ml） for 6h,12h,24h,and treated with TNF-α（3ng/ml） and PDTC（5mmol/ml） for 6h.The expression of eNOS gene was detected by RT-PCR.Results：RT-PCR confirmed that three groups cell lines all express eNOS but the levels were much lower in group 2 than the other two groups（P0.01）.ELISA confirmed that the level of TNF-α of serum was significantly higher in women with PE than in the normal pregnant women（P0.01）.eNOS expression level was down regulated by TNF-α in dose-depend and time-depend mode（P0.01）.The expression of eNOS was significantly higher in the cells which the PDTC and the TNF-α co-affected than in the cells which TNF-α affected lonely（P0.01）.Conclusion：TNF-α can down-regulated the expression of eNOS gene in HUVEC and PTDC inhibited the effect induced by TNF-α.This study laid experimental foundation for further research of the prevention and therapy of PTDC in PE.
出处
《中国优生与遗传杂志》
2011年第10期22-24,4,共4页
Chinese Journal of Birth Health & Heredity
