Quantitative assessment of cyanide in cystic fibrosis sputum and its oxidative catabolism by hypochlorous acid.
Quantitative assessment of cyanide in cystic fibrosis sputum and its oxidative catabolism by hypochlorous acid.
复制标题
囊性纤维化痰中氰化物的定量评估及其次氯酸的氧化分解代谢。
DOI：
10.1016/j.freeradbiomed.2018.09.007
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发表时间：
2018
影响因子：
7.4
通讯作者：
Cross,CarrollE
中科院分区：
文献类型：
--
作者：
Eiserich,JasonP;Ott,SeanP;Kadir,Tamara;Morrissey,BrianM;Hayakawa,KeriA;LaMerrill,MicheleA;Cross,CarrollE
基本原理 已知囊性纤维化 (CF) 患者会产生氰化物 (CN-)，尽管在确定总水平、精确的生物活性水平以及 CF 微生物群（尤其是 P.CN-）产生氰化物的特异性方面存在挑战。铜绿假单胞菌。我们的目标是通过一种简单而新颖的技术来测量 CF 痰中的总 CN 水平。铜绿假单胞菌阳性和阴性成年患者，检查 CN- 产生和降解的呼吸道 (RT) 机制，并检查痰中 CN-代谢物的翻译后蛋白质修饰情况。 方法通过使用市售的 CN-电极，测量添加 Cobinamide（一种对 CN- 具有极高亲和力的化合物）之前和之后的浓度，测定痰中 CN-浓度。通过蛋白质印迹法检测蛋白质氨甲酰化。测量和主要结果发现商业 CN 电极以高度可变的方式高估了 CF 痰液中的 CN 水平；钴酰胺的添加纠正了这个分析问题。虽然P.铜绿假单胞菌阳性患者往往具有较高的总 CN 值，阳性和阴性痰液中 CN 水平没有显着差异。炎症氧化剂次氯酸 (HOCl) 可快速分解 CN-，形成氯化氰 (CNCl) 和氨基甲酰化物质氰酸盐 (NCO-)。在 CF 痰中发现了氨甲酰化蛋白，与报道的哮喘中的发现类似。结论我们的研究表明，由于多种生物合成和代谢过程，CN-是发炎的 CF 气道中的短暂物种。 CN- 的稳定代谢物，例如氰酸盐或氨基甲酰化蛋白质，可能是 CF 气道中总 CN-产生的合适生物标志物。
RationaleCystic fibrosis (CF) patients are known to produce cyanide (CN-) although challenges exist in determinations of total levels, the precise bioactive levels, and specificity of its production by CF microflora, especiallyP. aeruginosa. Our objective was to measure total CN-levels in CF sputa by a simple and novel technique inP. aeruginosapositive and negative adult patients, to review respiratory tract (RT) mechanisms for the production and degradation of CN-, and to interrogate sputa for post-translational protein modification by CN-metabolites.MethodsSputa CN-concentrations were determined by using a commercially available CN-electrode, measuring levels before and after addition of cobinamide, a compound with extremely high affinity for CN-. Detection of protein carbamoylation was measured by Western blot.Measurements and main resultsThe commercial CN-electrode was found to overestimate CN-levels in CF sputum in a highly variable manner; cobinamide addition rectified this analytical issue. AlthoughP. aeruginosapositive patients tended to have higher total CN-values, no significant differences in CN-levels were found between positive and negative sputa. The inflammatory oxidant hypochlorous acid (HOCl) was shown to rapidly decompose CN-, forming cyanogen chloride (CNCl) and the carbamoylating species cyanate (NCO-). Carbamoylated proteins were found in CF sputa, analogous to reported findings in asthma.ConclusionsOur studies indicate that CN-is a transient species in the inflamed CF airway due to multiple biosynthetic and metabolic processes. Stable metabolites of CN-, such as cyanate, or carbamoylated proteins, may be suitable biomarkers of overall CN-production in CF airways.