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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >High urea and NaCl carbonylate proteins in renal cells in culture and in vivo, and high urea causes 8-oxoguanine lesions in their DNA.
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High urea and NaCl carbonylate proteins in renal cells in culture and in vivo, and high urea causes 8-oxoguanine lesions in their DNA.

机译：高浓度尿素和NaCl羰基化蛋白会在培养和体内的肾细胞中产生，而高尿素会在其DNA中引起8-氧鸟嘌呤损伤。

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摘要

Urea and NaCl are elevated in the renal inner medulla. We now find that a high concentration of urea or NaCl increases reactive oxygen species (ROS) in mouse renal inner medullary (mIMCD3) cells in culture. Previously, high NaCl, but not high urea, was found to cause DNA double-strand breaks. We now tested whether high urea or NaCl causes oxidative damage to DNA or cellular proteins. We find that high urea increases mIMCD3 cell DNA single-strand breaks and 8-oxoguanine lesions. High NaCl does not cause detectable 8-oxoguanine lesions. High urea or NaCl also greatly increases carbonylation of proteins in mIMCD3 cells. Carbonylation occurs within 5 min and with as little as 5 mM urea, a normal plasma level. It increases as urea is raised over the range in uremia. A high raffinose level increases ROS and carbonylation. High sorbitol and glycerol levels do not increase ROS or carbonylation. Carbonyl content is high in mouse renal inner medullas where interstitial NaCl and urea concentrations are normally high. There, numerous proteins are carbonylated, and carbonylation occurs in both collecting ducts and thin limbs. Conclusions: (i) Oxidative stress, associated with high urea, causes 8-oxoguanine DNA lesions in mIMCD3 cell DNA. (ii) High urea or NaCl carbonylates proteins in mIMCD3 cells and in renal inner medullary cells in vivo. (iii) In mIMCD3 cells a normal plasma concentration of urea causes carbonylation, and carbonylation increases over the uremic range of urea concentration, indicating that urea can contribute directly to the carbonylation found in uremia.
机译：尿素和氯化钠在肾内髓质中升高。现在，我们发现高浓度的尿素或氯化钠会增加培养物中小鼠肾内髓质（mIMCD3）细胞中的活性氧（ROS）。以前，发现高氯化钠而不是高尿素会导致DNA双链断裂。现在，我们测试了高尿素或氯化钠是否会引起DNA或细胞蛋白的氧化损伤。我们发现，高尿素会增加mIMCD3细胞DNA单链断裂和8-氧鸟嘌呤损伤。高氯化钠不会引起可检测的8-氧鸟嘌呤损伤。高尿素或氯化钠也大大增加了mIMCD3细胞中蛋白质的羰基化作用。羰基化发生在5分钟内，并且尿素低至5 mM（正常血浆水平）。随着尿素在尿毒症范围内的升高，它会增加。棉子糖含量高会增加ROS和羰基化。高的山梨糖醇和甘油含量不会增加ROS或羰基化。小鼠肾内髓质中羰基含量很高，而间质NaCl和尿素浓度通常很高。在那里，许多蛋白质被羰基化，羰基化同时发生在收集管和细肢中。结论：（i）与高尿素有关的氧化应激导致mIMCD3细胞DNA中的8-氧代鸟嘌呤DNA损伤。 （ii）体内mIMCD3细胞和肾内髓细胞中的高尿素或NaCl羰基化蛋白。 （iii）在mIMCD3细胞中，正常的尿素血浆浓度会导致羰基化，并且在尿毒症浓度的尿毒症范围内，羰基化会增加，这表明尿素可直接导致尿毒症中的羰基化。

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