Fig. 2: Plasma metabolic profiles of mice are altered by naproxen.

A The trial design. Eight week old mice of both sexes were randomly assigned to receive a diet containing naproxen or a chow diet control for 3 weeks prior to blood sampling. B Plasma tryptophan levels were compared between control and naproxen-fed mice (n = 16 per group; n denotes biological replicates) using two-sided 2 way Anova with Tukey’s multiple correction P < 0.0001. C Plasma kynurenine levels were compared between control and naproxen-fed mice (n = 16 per group; n denotes biological replicates) using two-sided 2-way Anova with Tukey’s multiple correction (P < 0.0001). D Plasma kynurenic acid levels were compared between control and naproxen-fed mice (n = 16 per group; n denotes biological replicates) using two-sided 2-way Anova with Tukey’s multiple correction. E Plasma xanthurenic acid levels were compared between control and naproxen-fed mice (n = 16 per group; n denotes biological replicates) using two-sided 2-way Anova with Tukey’s multiple correction. F Plasma anthranilic acid levels were compared between control and naproxen-fed mice (n = 16 per group; n denotes biological replicates). G ROC analysis of tryptophan: control vs. naproxen, (n = 16 per group, n denotes biological replicates), P < 0.0001. H ROC analysis of kynurenine: control vs. naproxen, P < 0.0005 (n = 16 per group, n denotes biological replicates), P = 0.0005. Plasma tryptophan levels were compared using two-way ANOVA with Tukey’s test among control mice on normal chow (n = 9; n denotes biological replicates), control mice on naproxen diet (n = 9; n denotes biological replicates), COX DKO mice on normal diet (n = 11; n denotes biological replicates), and COX DKO mice on naproxen diet (n = 7; n denotes biological replicates). J Comparison of change in tryptophan levels in control mice and COX-DKO mice (n = 9; n denotes biological replicates) on control chow or dietary naproxen (n = 7; n denotes biological replicates) using two sided Mann Whitney test. K Spearman correlation plot of urinary prostaglandin metabolites with plasma tryptophan and kynurenine in healthy volunteers (n = 16; n denotes biological replicates). The color bar shows Spearman coefficient (r) values. No significant correlations were found between tryptophan or kynurenine and urinary metabolites (P > 0.05). L Plasma tryptophan levels in Cox-1 knockout mice (n = 5; n denotes biological replicates) and Cox-2 knockout mice (n = 5; n denotes biological replicates) showed no significant differences compared to their respective cre -/- controls (n = 12 and n = 10; n denotes biological replicates), based on one-way ANOVA with Tukey’s test (P = 0.81 and P = 0.91, respectively). M Plasma kynurenine levels of COX-1 knockout mice (n = 5; n denotes biological replicates) and COX-2 knockout mice (n = 5; n denotes biological replicates) showed no significant differences compared to their respective cre -/- controls (n = 12 and n = 10; n denotes biological replicates), based on one-way ANOVA with Tukey’s test (P = 0.97 and P = 0.37, respectively). N Plasma tryptophan levels were compared among control mice on chow (n = 5; n denotes biological replicates), control mice on PPA (n = 16; n denotes biological replicates), DKO mice on chow (n = 24; n denotes biological replicates), and DKO mice on PPA (n = 14 n denotes biological replicates), using Kruskal-Wallis with Dunn’s test. No significant difference was found between control and DKO mice on PPA diet (P = 0.9). O Plasma kynurenine levels were compared among control mice on chow (n = 5; n denotes biological replicates), control mice on PPA (n = 16; n denotes biological replicates), DKO mice on chow (n = 24; n denotes biological replicates), and DKO mice on PPA (n = 14; n denotes biological replicates), using Kruskal-Wallis with Dunn’s test. No significant difference was found between control and DKO mice on the PPA diet (P = 0.9). P Plasma kynurenine/tryptophan ratio were compared among control mice on chow (n = 5; n denotes biological replicates), control mice on PPA (n = 16; n denotes biological replicates), DKO mice on chow (n = 24; n denotes biological replicates), and DKO mice on PPA (n = 14; n denotes biological replicates), using Kruskal-Wallis with Dunn’s test. No significant difference was found between control and DKO mice on the PPA diet (P = 0.9). The green, blue, purple and black circles denote the control mice on the naproxen diet, control mice on the PPA diet, Cox-DKO mice on the PPA diet, and Cox-DKO mice on the control chow diet, respectively. All the data are expressed in mean ± SD *Designates statistical significance, **p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.