肿瘤坏死因子α和血管紧张素Ⅱ干预内皮细胞后细胞凋亡和诱导型一氧化氮合酶的表达
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云南省科委(2001C0070M)资助项目


The Study on Inducible Nitric Oxide Synthase Expression and Cell Apoptosis treated by Tumor Necrosis Factor-α and Angiotensin Ⅱ
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    摘要:

    目的探讨肿瘤坏死因子α和血管紧张素Ⅱ在导致内皮细胞凋亡过程中诱导型一氧化氮合酶的表达变化及核因子κB的作用。方法核因子κB抑制剂吡咯烷二硫代氨基甲酸盐预处理和未预处理原代培养的脐静脉内皮细胞,用肿瘤坏死因子α和血管紧张素Ⅱ分别进行干预,逆转录聚合酶链反应检测诱导型一氧化氮合酶mR-NA的表达,免疫印迹法检测诱导型一氧化氮合酶和IκBα的蛋白表达,电泳迁移率分析检测核因子κB的活性,TUNEL法检测细胞凋亡。结果在10μg/L肿瘤坏死因子α和1μmol/L血管紧张素Ⅱ的干预下,核因子κB的活性显著增加(p<0.05),诱导型一氧化氮合酶mRNA和蛋白的表达与对照组比较显著增加(p<0.05),细胞凋亡发生显著增加(p<0.05);吡咯烷二硫代氨基甲酸盐抑制肿瘤坏死因子α和血管紧张素Ⅱ引起的细胞凋亡和诱导型一氧化氮合酶表达的增加。结论在肿瘤坏死因子α和血管紧张素Ⅱ作用于内皮细胞时,通过降解IκBα引起诱导型一氧化氮合酶的核转位,后者可引起诱导型一氧化氮合酶表达上调和细胞凋亡。

    Abstract:

    Aim To explore the cell apoptosis and inducible nitric oxide synthase(iNOS) expression in the cultured endothelial cells induced by tumor necrosis factor(TNF-α) and angiotensin Ⅱ(AngⅡ). Methods The cultured endothelial cells were treated with TNF-α(10 μg/L) and AngⅡ(1 μmol/L) in absence and presence of pyrroledithiocarbomate(PDTC);the mRNA of iNOS was measured by reverse transcriptionpolymerase chain reaction(RT-PCR),the protein of iNOS and IκBα were assessed by Western blotting,the activity of nuclear factor-κB(NF-κB) was evaluated by electrophoretic mobility shift assay(EMSA),and the cell apoptosis was detected with TUNEL. Ruselts The mRNA and protein of iNOS were significantly increased in the endothelial cells induced by TNF-α and AngⅡ(p<0.05) and PDTC could prevent this increase;at the same time,TNF-α and AngⅡ could increase the apoptosis of cells(p<0.05) and PDTC could prevent this increase. Conclusion TNF-α and AngⅡ induced the up-regulation of iNOS and the cell apoptosis,and the NF-κB play a key role in this process.

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杨丽霞,郭瑞威,齐峰,石燕昆,王红.肿瘤坏死因子α和血管紧张素Ⅱ干预内皮细胞后细胞凋亡和诱导型一氧化氮合酶的表达[J].中国动脉硬化杂志,2006,14(10):849~852.

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  • 收稿日期:2006-03-27
  • 最后修改日期:2006-10-08
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