基础医学与临床 ›› 2014, Vol. 34 ›› Issue (3): 386-390.

• 研究论文 • 上一篇    下一篇

高磷对维持性血液透析透患者微炎性反应和氧化应激的影响及机制

陈艳红1,陈星2,成梅初2,袁芳3,陈俊香2,刘伏友3   

  1. 1. 湖南省人民医院
    2. 中南大学湘雅二医院
    3. 中南大学湘雅二医院肾内科
  • 收稿日期:2013-06-14 修回日期:2013-09-29 出版日期:2014-03-05 发布日期:2014-02-27
  • 通讯作者: 陈星 E-mail:375811942@qq.com

The effect and mechanism of phosphorus concentration on microinflammatory and oxidative stress response in maintenance hemodialysis patients

  • Received:2013-06-14 Revised:2013-09-29 Online:2014-03-05 Published:2014-02-27

摘要: 目的 探讨高磷对维持性血透患者微炎性反应及氧化应激的影响及机制。方法 实验分A组(正常人对照组),B组(维持性血透患者组即Maintenance hemodialysis,MHD),C组(不同磷酸二氢钠浓度剂量组 1.5mmol/L、 C2 2.5mmol/L及C3 5.5mmol/L),D组(5.5mmol/L磷酸二氢钠作用不同时间组D1 6h、D2 12h及D3 24h)和E组(无磷酸二氢钠作用空白组E1 6h、E2 12h及E3 24h组)。用Real Time PCR及Western blot分别检测外周血单个核细胞NF-κBP65及NOX2/gp91的表达。ELISA检测血浆及细胞培养液上清IL-6含量,硫氮巴比妥酸法检测MDA含量。结果 MHD组PBMC中NF-κBP65、NOX2 mRNA及Phospho-NF-κBP65、NOX2 蛋白表达及血浆IL-6、MDA水平明显高于正常组;磷酸二氢钠作用下,培养液上清IL-6、MDA产生呈剂量、时间依赖性增加(p<0.05)。NF-κBP65mRNA、Phospho-NF-κBP65蛋白和NOX2/gp91 mRNAN及蛋白水平表达呈剂量、时间依赖性上调(p<0.05);NF-κBP65mRNA及Phospho-NF-κBP65蛋白表达与培养液上清IL-6产生呈正相关, NOX2/gp91mRNA及蛋白表达与培养液上清MDA产生呈正相关。结论 高磷可能通过活化NF-κBP65、NOX2/gp91信号通路介导炎症介质、氧化应激产物产生增加,高磷血症可能为MHD患者微炎性反应状态和氧化应激状态的危险因素。

关键词: 维持性血液透析, 高磷血症, 微炎症, 氧化应激

Abstract: Objective To investigate the effect of high phosphate on mRNA and protein expression of intracellular NF-κB and NOX2/gp91 in PBMC of MHD patients and serum inflammation mediators and MDA production,so as to understand the possible relationship among hyperphosphatemia, micro-inflammatory states and oxidative stress. Method They were divided into five groups: control, MHD, dose, time and blank groups.Western blot and Real-time PCR were respectively used to determine the mRNA and protein expression of NF-κBP65、NOX2.The concentration of IL-6 in the culture supernatant was quantified by the ELISA kit and the TBA assay kit.Results NF-κB and NOX2 signal pathway in PBMCs show abnormal activation status in MHD patients. Both expression of NOX2/gp91 mRNA and protein levels increased dose-and time-dependent manners totally. IL-6 and MDA production in supernatant up-regulate dose-and time-dependent manners totally. NF-κB P65 mRNA and Phospho-NF-κBP65 protein were positively correlated with IL-6 production. NOX2/gp91 mRNA and protein expression were positively correlated with MDA production. Conclusion High phosphate could increase the production of supernatant IL-6 and MDA by activating NF-κB and NOX2 signaling pathway in PBMCs, thus could be a risk factor for microinflammatory and oxidative stress status of MHD patients.

Key words: maintenance hemodialysis, hyperphosphatemia, microinflammatory, oxidative stress

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