Volume 118, Issue 6, Pages 1140-1148 (June 2000) Enhanced urinary excretion of cysteinyl leukotrienes in patients with acute alcohol intoxication  Masahito Uemura, Wolf-dieter Lehmann, Winfried Schneider, Helmut K. Seitz, Axel Benner, Andrea Keppler-Hafkemeyer, Peter Hafkemeyer, Hideyuki Kojima, Masao Fujimoto, Tadasu Tsujii, Hiroshi Fukui, Dietrich Keppler  Gastroenterology  Volume 118, Issue 6, Pages 1140-1148 (June 2000) DOI: 10.1016/S0016-5085(00)70367-2 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 Catabolic pathway of LTE4 and N-acetyl-LTE4 via ω-oxidation. 20-Hydroxylation of LTE4 and N-acetyl-LTE4 is catalyzed by microsomal LTE4 20-monooxygenase,16 followed, particularly in hepatocytes, by 2 dehydrogenase-catalyzed reactions leading to 20-carboxy-LTE4 and 20-carboxy-N-acetyl-LTE419 (the ω-aldehyde intermediate is not shown). LTB4 is ω-oxidized in reactions that are analogous.14,18 Thedashed double lines indicate the ethanol-induced block in this degradation pathway. Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 Urinary excretion of LTE4, N-acetyl-LTE4, LTB4, and 20-hydroxy-LTB4 in normal subjects and patients with alcohol intoxication. Excretion of LTE4 and N-acetyl-LTE4 was determined by HPLC separation and subsequent RIA. Urinary excretion of LTE4 and N-acetyl-LTE4 was 8-10 times higher in patients with alcohol intoxication than normal subjects. LTB4 and 20-hydroxy-LTB4 were identified and quantified in a limited number of patients by HPLC separation and subsequent GC-MS. LTB4 and 20-hydroxy-LTB4 were below the detection limit in all samples from normal subjects and in most patients with alcohol intoxication. The lines above normal subjects in the columns for LTB4 and 20-hydroxy-LTB4 indicate the detection limits for normal individuals. Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Changes in urinary cysteinyl LTs during the decrease of blood ethanol concentration in patients with alcohol intoxication. Urinary LTE4 and N-acetyl-LTE4 levels significantly decreased when the blood ethanol concentration declined and were below detection limit. The hatched areas indicate the normal range of cysteinyl LTs (mean ± 2 SD from normal subjects). NAc, N-acetyl. Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 Relationship between urinary cysteinyl LTs and standard liver function tests. There was an increase in the excretion of cysteinyl LTs (LTE4 + N-acetyl-LTE4) from lowest in healthy subjects to alcoholic patients with normal liver function test results (P < 0.001), and to highest in alcoholic patients with mild abnormalities of liver function tests (P < 0.001). Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Mass spectrometric analysis of LTB4 and 20-hydroxy-LTB4 by negative ion chemical ionization. A 20-mL sample of urine was spiked with 50 ng each of [18O2]LTB4 and 20-hydroxy-[18O2]LTB4. LTB4 and 20-hydroxy-LTB4 were separated by RP-HPLC and subsequent SP-HPLC, derivatized, and analyzed by GC-MS, as described in Patients and Methods. (A) Patient A2. In the m/z 487 of the upper chromatogram, a peak was recognized that is consistent with the presence of LTB4 and corresponds to the internal standard at the retention time of 16 minutes 4 seconds in the m/z 491 of the lower chromatogram. (B) Patient B10. In the m/z 575 of the upper chromatogram, a peak indicates the presence of 20-hydroxy-LTB4 with the internal standard at the retention time of 19 minutes 35 seconds in the m/z 579 of the lower chromatogram. The vertical scale is identical for the corresponding traces of samples and internal standards. The peaks of only the internal standard of LTB4 and 20-hydroxy-LTB4 at m/z 487 and m/z 575, respectively (lowest column), were apparently small as compared with the sharp and significant peaks obtained from the urine samples in patients with alcohol intoxication. Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 6 Blood ethanol and urinary LTE4 concentrations after ethanol consumption in healthy volunteers. They consumed 60 g ethanol for 1 hour. (A) Blood ethanol concentrations were the highest 1 hour after intake, thereafter gradually decreased, but were still high 5 hours after ethanol intake. (B) LTE4 concentrations were significantly higher 3-5 hours after ethanol consumption compared with basal values (3, 4, and 5 hours; P < 0.01, P < 0.001, and P < 0.001, respectively). (C) LTE4/creatinine ratios were significantly higher 3 and 4 hours after ethanol consumption compared with basal values (3 and 4 hours; P < 0.05 and P < 0.001, respectively). Gastroenterology 2000 118, 1140-1148DOI: (10.1016/S0016-5085(00)70367-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions