Measurement of urinary desmosine by isotope dilution and high performance liquid chromatography. Correlation between elastase-induced air-space enlargement in the hamster and elevation of urinary desmosine.
Measurement of urinary desmosine by isotope dilution and high performance liquid chromatography. Correlation between elastase-induced air-space enlargement in the hamster and elevation of urinary desmosine.
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DOI：
10.1164/ajrccm/144.2.284
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发表时间：
1991-08
期刊：
影响因子：
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通讯作者：
P. Stone;J. Bryan-Rhadfi;E. Lucey;D. Ciccolella;George Crombie;B. Faris;G. Snider;C. Franzblau
中科院分区：
文献类型：
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作者：
P. Stone;J. Bryan-Rhadfi;E. Lucey;D. Ciccolella;George Crombie;B. Faris;G. Snider;C. Franzblau
用于测量弹性蛋白特异性交联、锁链素（DES）和异锁链素（IDES）的方法的准确性受到质疑，因为尿液中的污染物可能导致值升高。在本研究中，尿样加标已知量的[14 C]DES，并在6 N HCl中回流。水解尿的Sephadex G-15色谱用于去除污染物。DES和IDES通过高效液相色谱法（HPLC）以及氨基酸分析进行定量。回收的同位素量用于确定整个程序期间的损失，同位素稀释用于计算尿液中最初存在的内源性DES和IDES的量。由于两种方法获得的值相似，因此所有后续分析均使用更快速的HPLC法。在一个实验中，在用人中性粒细胞弹性蛋白酶进行肠道内治疗后3天，从仓鼠收集的尿液中DES的量（300微克）或猪胰弹性蛋白酶（300微克）为0.212 +/- 0.012（平均值+/- SEM，对单个池进行两次测量）和0.816 +/- 0.005（两次测量）微克/仓鼠/天。对照仓鼠尿液的平均值为0.074 +/- 0.008（8次测量）μ g/仓鼠/天。HNE和PPE处理的仓鼠的平均线性截距值分别为对照值的119和159%，这表明空域大小的增加与尿DES的升高呈正相关。（250字处删节）
The accuracy of methods employed to measure the elastin-specific crosslinks, desmosine (DES) and isodesmosine (IDES), has been called into question because contaminants in the urine may cause elevated values. In the present study urine samples were spiked with a known amount of [14C]DES and refluxed in 6 N HCl. Sephadex G-15 chromatography of the hydrolyzed urine employed to remove contaminants. DES and IDES were quantified by high performance liquid chromatography (HPLC) as well as by amino acid analysis. The amount of isotope recovered was used to determine losses during the overall procedure and the isotope dilution to calculate the amounts of endogenous DES and IDES originally present in the urine. Because similar values were obtained by both methods, the more rapid HPLC method was used for all succeeding analyses. In one experiment, the DES amounts in urine collected from hamsters for 3 days after intratracheal treatment with human neutrophil elastase (300 micrograms) or porcine pancreatic elastase (300 micrograms) were 0.212 +/- 0.012 (mean +/- SEM, two measurements on a single pool) and 0.816 +/- 0.005 (two measurements) microgram per hamster per day, respectively. Urine from control hamsters had a mean value of 0.074 +/- 0.008 (eight measurements) microgram per hamster per day. The HNE- and PPE-treated hamsters had mean linear intercept values of 119 and 159% of control values, respectively, giving a positive correlation between increase in airspace size and elevation of urinary DES.(ABSTRACT TRUNCATED AT 250 WORDS)