Extracellular DNA Affects NO Content in Human Endothelial Cells
Extracellular DNA Affects NO Content in Human Endothelial Cells
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DOI：
10.1007/s10517-010-0906-3
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发表时间：
2010-08-01
影响因子：
0.7
通讯作者：
Veiko, N. N.
中科院分区：
文献类型：
--
作者：
Efremova, L. V.;Alekseeva, A. Yu.;Veiko, N. N.
由于细胞凋亡和可能的从头DNA合成，细胞外DNA片段永久释放到血流中。为了解细胞外DNA是否影响血管紧张度调节因子一氧化氮（NO）的合成，我们研究了3种不同序列的人工DNA探针和10份来自健康人、高血压和动脉粥样硬化患者的细胞外DNA对培养的内皮细胞（HUVEC）NO合成的影响。为了检测活细胞和培养液中的NO，我们使用了NO特异性试剂CuFL渗透到细胞中，并形成荧光产物FL-NO后与NO的相互作用。人类基因组DNA片段影响内皮细胞中的NO含量，这种效果依赖于DNA片段的碱基序列和浓度。将人工DNA和来自健康人的细胞外DNA以低浓度（5 ng/ml）添加到细胞培养物中使检测到的NO浓度最多增加4倍。富含胞嘧啶鸟嘌呤（CG）的核糖体重复序列的转录序列的片段是最强大的NO诱导剂。DNA片段对NO合成的影响与低剂量氧化剂H2 O2和17 β-雌二醇的影响相当。与对照组相比，从高血压和动脉粥样硬化患者中获得的细胞外DNA样品使细胞和培养基中的NO含量降低了1.3-28倍;该效果与富含CG的序列的含量相关。
Fragments of extracellular DNA are permanently released into the blood flow due to cell apoptosis and possible de novo DNA synthesis. To find out whether extracellular DNA can affect the synthesis of nitric oxide (NO), one of key vascular tone regulators, we studied in vitro effects of three artificial DNA probes with different sequences and 10 samples of extracellular DNA (obtained from healthy people and patients with hypertension and atherosclerosis) on NO synthesis in endothelial cell culture (HUVEC). For detection of NO in live cells and culture medium, we used a NO-specific agent CuFL penetrating into the cells and forming a fluorescent product FL-NO upon interaction with NO. Human genome DNA fragments affected the content of NO in endothelial cells; this effect depended on both the base sequence and concentration of DNA fragments. Addition of artificial DNA and extracellular DNA from healthy people into the cell culture in a low concentration (5 ng/ml) increased the detected NO concentration by 4-fold at most. Cytosine-guanine (CG)-rich fragment of the transcribed sequence of ribosomal repeat was the most powerful NO-inductor. The effect of DNA fragments on NO synthesis was comparable with that of low doses of oxidizing agents, H2O2 and 17 beta-estradiol. Extracellular DNA samples obtained from patients with hypertension and atherosclerosis decreased NO content in cells and medium by 1.3-28 times compared to the control; the effect correlated with the content of CG-rich sequences.