A Liquid Chromatography-Quadrupole Time-of-Flight (LC-QTOF)-based Metabolomic Approach Reveals New Metabolic Effects of Catechin in Rats Fed High-Fat Diets Anthony Fardet,† Rafael Llorach,‡ Jean-Franc¸ois Martin,† Catherine Besson,† Bernard Lyan,† Estelle Pujos-Guillot,† and Augustin Scalbert*,† UMR 1019, Unite´ de Nutrition Humaine, INRA, Centre de Recherche de Clermont-Ferrand/Theix, F-63122 St-Gene`s-Champanelle, France, and Nutrition and Food Science Department, XarTA, Pharmacy School, University of Barcelona, 08028 Barcelona, Spain Received January 15, 2008
Unbalanced diets generate oxidative stress commonly associated with the development of diabetes, atherosclerosis, obesity and cancer. Dietary flavonoids have antioxidant properties and may limit this stress and reduce the risk of these diseases. We used a metabolomic approach to study the influence of catechin, a common flavonoid naturally occurring in various fruits, wine or chocolate, on the metabolic changes induced by hyperlipidemic diets. Male Wistar rats (n ) 8/group) were fed during 6 weeks normolipidemic (5% w/w) or hyperlipidemic (15 and 25%) diets with or without catechin supplementation (0.2% w/w). Urines were collected at days 17 and 38 and analyzed by reverse-phase liquid chromatography-mass spectrometry (LC-QTOF). Hyperlipidic diets led to a significant increase of oxidative stress in liver and aorta, upon which catechin had no effect. Multivariate analyses (PCA and PLS-DA) of the urine fingerprints allowed discrimination of the different diets. Variables were then classified according to their dependence on lipid and catechin intake (ANOVA). Nine variables were identified as catechin metabolites of tissular or microbial origin. Around 1000 variables were significantly affected by the lipid content of the diet, and 76 were fully reversed by catechin supplementation. Four variables showing an increase in urinary excretion in rats fed the high-fat diets were identified as deoxycytidine, nicotinic acid, dihydroxyquinoline and pipecolinic acid. After catechin supplementation, the excretion of nicotinic acid was fully restored to the level found in the rats fed the low-fat diet. The physiological significance of these metabolic changes is discussed. Keywords: Catechin • antioxidants • high-fat diets • metabolomics • urine • rats
Introduction Major diet-related diseases or metabolic disorders such as atherosclerosis, cancers, diabetes and obesity are associated with increased oxidative stress.1–9 Unbalanced diets (hyperlipidemic, hyperglycemic or hypercaloric) are known risk factors for such diseases. They increase oxidative stress in rats and humans, both in postprandial10–12 and chronic states.13,14 The levels of oxidized lipids (thiobarbituric acid reactive substances, TBARS) and of several antioxidant enzymes (superoxide dismutase, SOD, and glutathione (GSH)-reductase/ transferase) were, respectively, increased and reduced in various tissues upon the intake of high-sucrose and high-fat diets.10,11,15,16 GSH depletion was observed in the aorta of apoEdeficient mice spontaneously developing atherosclerosis.17 Antioxidant polyphenols are able to counteract oxidative stress generated by such unbalanced diets.18,19 Flavonoids such as quercetin or catechins were shown to reduce pro-oxidant * To whom correspondence should be addressed. Tel: +33(0)473624787. Fax: +33(0)473624638. E-mail:
[email protected]. † Unite´ de Nutrition Humaine. ‡ University of Barcelona.
2388 Journal of Proteome Research 2008, 7, 2388–2398 Published on Web 05/17/2008
effects of high-fat or high-sucrose diets in the rat.20,21 These effects might be explained by their free-radical scavenging properties, but more recent studies also suggest that they may control the activation/repression of transcription factors and modulate the expression of key genes involved in the control of the cell redox status, such as NADPH-quinone oxidoreductase, glutathione S-transferase or ferritin.22–24 Thus, metabolic effects of polyphenols cannot be reduced to their sole freeradical scavenging capacity and antioxidant effects. More diverse metabolic effects were associated with a reduction of the risk for various chronic diseases.25,26 Metabolic responses to unbalanced diets and to protective phytochemicals such as polyphenols are therefore complex and not all related to the control of the cell redox state. The complexity of these effects can be explored by metabolomics, which has been defined as the quantitative measurement of the multivariate metabolic responses of multicellular systems to pathophysiological stimuli or genetic modification.27 Itinferstheidentificationandquantificationofallmetabolitessthe metabolomesin the considered biological system, most often by NMR and MS techniques. In particular, the higher sensitivity 10.1021/pr800034h CCC: $40.75
2008 American Chemical Society
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Metabolic Effects of Catechin in Rats Fed High-Fat Diets a
Table 1. Composition of Control and Hyperlipidemic Diets (%, w/w)
LF diet (5% lipids)
MF diet (15% lipids)
HF diet (25% lipids)
Casein Starch Lards Peanut oil Colza oil Cellulose Vitaminsb Mineralsc Bitartrate choline Vitamin E (mg)d Energy (kcal/100 g)
20 65.25 3.5 0.75 0.75 5 1 3.5 0.25 12 402
20 55.25 10.5 2.25 2.25 5 1 3.5 0.25 12 450
20 45.25 17.5 3.75 3.75 5 1 3.5 0.25 12 498
a LF, low-fat diet (5% lipids); MF, medium-fat diet (15% lipids); HF, high-fat diet (25% lipids). b In order to reach a total of 12 mg vitamin E/ 100 g of diet, we used a mixture of two vitamin mixes: one with vitamin E and one without; vitamin mixture (g/kg mix) contains thiamin (1.24), riboflavin (0.75), pyridoxine (0.7), nicotinic acid (3), pantothenate (1.6), folic acid (0.2), biotin (1), cyanocobalamin (2.5), retinyl palmitate (1.24), cholecalciferol (0.2), phylloquinone (1.5), and choline (0.23). c Mineral mixture is AIN-93G, MP Biomedicals, Inc. d The content of vitamin E in both oils and lards was taken into consideration.
of the latter led to their rapid development in metabolomics applications, with the routine quantification of hundreds or thousands of different metabolites present in the micromolar range or below.28 Subtle metabolic changes characterizing the early stages of chronic diseases and their interaction with the diet or various dietary factors have thus been described using metabolomics approaches.29–31 These approaches have been used to characterize the metabolic effects associated with caloric restriction,32,33 vitamin deficiency34 or intake of PUFA-rich oils and of antioxidant-rich foods such as soy, chamomile, and tea.35–39 The wealth of information collected in metabolomics has revealed subtle differences in metabolic profiles resulting from the consumption of closely related foods: whole-grain and refinedgrain cereals,40 different PUFA-containing oils36 or green and black tea.35 Finally, metabolomics was also applied to explore the metabolic effects of pure dietary antioxidants such as epicatechin.41 In the present work, the ability of catechin, a major flavonoid found in many fruits, wine, tea or cocoa, to counteract metabolic changes induced by high-fat diets is studied in the rat. Urinary metabolic profiles measured by highresolution mass spectrometry were compared. Evidence of the reversion of some of the metabolic effects of high-fat diets by catechin is provided.
Materials and Methods Animals and Diets. Six groups of male Wistar rats (n ) 8/group) weighing 324 g (range 284-379 g) were fed during 6 weeks (day 0-42) normo-(5%) or hyperlipidemic (15 and 25%) diets equilibrated for vitamin E, and supplemented or not with 0.2% (+)-catechin. The 0.2% supplementation correspond to the 1 g of polyphenol consumed daily (on a dry matter basis)
by human and adapted to rats. Compositions of the diets are given in Table 1. Lipids were 70% lards, 15% peanut oil and 15% colza oil which corresponded to a ratio polyunsaturated/ monounsaturated/saturated lipids of 17/50/33. Vitamin mix was AIN-93VX (UPAE, INRA, Jouy-en-Josas, France) and mineral mix was AIN-93G (MP Biomedicals).42 Animals were allowed free access to 30 g of fresh food per day, and to tap water ad libitum. Rats were first housed 2 per cage during the first 2 weeks, and then transferred to metabolic cages (1 per cage) during the 4 last weeks, and maintained in a temperaturecontrolled room (22 °C), with a dark period from 2000 to 0800 h. They were handled according to the recommendations of the Institutional Ethics Committee (Clermont-Ferrand University). The body weight of rats was recorded each week, and food intake twice a week. Urine samples were collected twice a day (1600-0900 h and 0900-1600 h) at days 17 and 38, and stored at -20 °C. Sampling Procedure. Rats were anesthetized at day 42, at the end of the dark period with sodium pentobarbital (40 mg/ kg) and maintained at 37 °C during sample collection. An abdominal incision was made, and blood was withdrawn from the abdominal aorta in heparinized tubes and centrifuged at 1400g for 10 min. The supernatant (plasma) was collected and stored at -80 °C. Heart, liver and aorta were also collected. Heart and liver were flushed free of blood by injection of phosphate buffer saline. Tissues were weighted and freezedclamped into liquid nitrogen and stored at -80 °C. Lipemia and Antioxidant Status Measurements. Plasma triglycerides and total cholesterol were determined enzymatically (Triglycerides PAP 150 & Cholesterol RTU, BioMe´rieux, Marcy-l’Etoile, France). Ferric reducing ability of plasma (FRAP) was determined as described previously.43 TBARS in urine, heart and liver samples were estimated according to Lee et al.44 Thiobarbituric acid reactive substances (TBARS) were measured in urine collected between 1600 and 0900 h at day 38, and expressed as nanomole equivalents of MDA. For total GSH concentration measurement, frozen heart and liver were finely grounded, homogenized and sonicated during 30 s in 2% perchloric acid solution containing 5 mM EDTA. Aorta were ground with an Ultraturax during 30-60 s in 2% perchloric acid solution containing 5 mM EDTA. After centrifugation (14 000g for 15 min), and filtration through 0.22 µm-filter (Millex, Millipore Corporation, Bedford, MA) for aorta supernatants, total GSH concentration was measured using a standard enzymatic recycling procedure as described previously by Robinson et al.45 The activities of liver enzymes related to glutathione metabolism were measured in an aliquot of liver powder homogenized in 75 mM phosphate buffer, 2 mM dithiothreitol, and 0.3 M sucrose, sonicated for 30 s, centrifuged (14 000g for 15 min), and filtered through 0.22 µm-filter (Millex, Millipore Corporation, Bedford, MA). Glutathione reductase (EC 1.6.4.2) was measured in the presence of GSSG by following the oxidation of NADPH.46 Glutathione peroxidase (EC 1.11.1.9) was measured in the presence of GSH and H2O2 by following
Table 2. Identified Urinary Endogenous Metabolites Significantly Affected by the Catechin and/or Lipid Content in the Diet at day 38 (two-way ANOVA) metabolite
RT (min)
detected mass m/z [M + H]+
theoretical mass m/z [M + H]+
MS/MS fragments
mass difference (mDa)
Pdiet
Pcatechin
Pinteraction
Deoxycytidine Nicotinic acid Dihydroxyquinoline Pipecolinic acid
1.6 2.3 7.4 1.7
228.1008 124.0409 162.0570 130.0880
228.0979 124.0393 162.0550 130.0862
112/95/69 106/80/78 144/116/89 84
3.1 1.6 2.0 1.8
0.391 0.785 0.016 0.025
0.918 0.851 0.001 0.351
0.043 0.009 0.152 0.125
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Table 3. Effect of Lipid and Catechin Content in the Diet on Oxidative Stress and Lipid Biomarkers in Rats LF diet
Liver GSH (µmol/g liver) Aorta GSH (µmol/g aorta) Liver GSH-peroxidase (U/g liver) Liver GSH-reductase (U/g liver) Liver GSH-transferase (U/g liver) Urine MDA (nmol/17 h) Liver MDA (nmol/g liver) Heart MDA (nmol/g heart) Plasma FRAP (µmol Fe/L) Plasma cholesterol (mM) Plasma triglycerides (mM)
MF diet
NP
PP
8.2 ( 0.3
7.6 ( 0.2
a
a
HF diet
NP
PP
7.1 ( 0.4
6.5 ( 0.4
b
Two-way ANOVA
NP
PP
6.8 ( 0.2
6.6 ( 0.2
Pdiet
b
Pcatechin Pinteraction
0.0004 0.0700
0.7615
0.73 ( 0.04 0.71 ( 0.02 a 0.56 ( 0.04 0.57 ( 0.06 b
0.50 ( 0.03 0.53 ( 0.02 b 0.8) with the lipid content of the diets whatever the sampling time and the presence or absence of catechin. Among all variables affected by the lipid content in the diet, some were tentatively identified by comparing experimental mass with theoretical mass (∆mDa e 5 mDa) of the metabolites given in HMDB: deoxycytidine (m/z 228), nicotinic acid (m/z 124), 2,4-dihydroxyquinoline (m/z 162) and pipecolinic acid (m/z 130). Their identities were confirmed by comparison of their retention time and MS-MS fragmentation pattern to that of standards (see Materials and Methods) (Table 2 and Figure 4). The other variables could not be identified, either due to the lack of plausible metabolites with closely related mass in common metabolite databases (see Materials and Methods) or to the absence of available standards when putative identification was suggested by comparison of exact masses with those registered in these databases. 2. Effect of the Catechin Supplementation on the Urinary Metabolome. Three-way ANOVA showed that 1781 variables (32% of the 5587 variables filtered) are significantly affected by the supplementation of catechin, interaction effect with lipid content and sampling time included. Several clusters of ions of high intensity correspond to catechin and its metabolites. Catechin metabolites could be identified on the basis of their expected exact mass and characteristic fragments formed in the electrospray source: loss of a glucuronide moiety (m/z 176) for glucuronide derivatives, loss of a sulfate moiety (m/z 80) for sulfated derivatives and formation of typical fragments for catechin, catechin glucuronide (m/z 165/139/123) and methylcatechin (m/z 179/139/137) metabolites56 (Table 4). The following metabolites of catechin could thus be recognized: methylcatechin, catechin glucuronide, methylcatechin glucuronide, hydroxyphenylvalerolactone glucuronide, methoxyhydroxyphenylvalerolactone, methoxyhydroxyphenylvalerolactone glucuronide, and sulfated methoxyhydroxyphenylvalerolactone. Identity of catechin, hippuric acid and p-hydroxyhyppuric acid was confirmed by comparison with authentic standards. 3. Metabolic Interactions between Catechin Supplementation and High-Fat Diets. Interactions between catechin and lipids were characterized by mixed model analysis (see Materials and Methods). The intensity of 85 variables affected by an increase of the lipid content in the diet (P < 0.05) was reversed by catechin supplementation, at day 17 and/or day 38, and for MF and/or HF diets. This concerned a total of 76 different Journal of Proteome Research • Vol. 7, No. 6, 2008 2393
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Figure 4. Evolution of the spectral intensity of 2′-deoxycytidine (A), nicotinic acid (B), dihydroxyquinoline (C) and pipecolinic acid (D) in urine of rats after feeding low-fat (LF), medium-fat (MF) and high-fat (HF) diets without catechin at day 38. Values are means ( SEM (n ) 6 rats/group); different letters indicate significant difference (P < 0.05).
markers. Twenty-eight markers were reversed at day 17 and 50 at day 38. One of these markers was identified as nicotinic acid (P < 0.05 for “lipid × catechin” interaction at day 38 for MF diet) (Figure 5, Table 2). Although the variable identified as deoxycytidine exhibited a significant “diet × catechin” interaction, it did not have a reversion profile as described in Materials and Methods.
Discussion The metabolic effects of polyphenols and other antioxidants are commonly assessed in clinical trials by measuring plasmatic concentrations of lipids, glucose or of some markers of oxidative stress.57 However, results are often inconsistent and do not reflect the diversity of polyphenol metabolic effects. In particular polyphenols like catechins are thought today to be more than antioxidants acting as free radical scavengers.25 Recent investigations have shown that flavonoids can modulate several cell signaling pathways involved in the control of the expression of genes coding for antioxidant or detoxifying enzymes.58,59 Several flavonoids can trigger Nrf2 (transcription factor)-ARE (Antioxidant Responsive Element) and MAP Kinase signaling pathway.24,60–62 Such modulation of gene expression by flavonoids have various consequences, for example, on inflammation, oxidative or cell proliferation.63 Metabolomics, combining high-throughput analytical methods and multivariate statistical analyses, allows the characterization of the effects of diets or nutrients on the metabolism under a holistic aspect. Such an approach is expected to provide new insights on mechanisms of action and to identify new markers of effects.29,64 In the present work, rats were fed high-fat diets known to induce oxidative stress, supplemented or not with catechin. The effects of catechin were measured using both common biochemical markers and a metabolomic approach applied to urine. 2394
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Hyperlipidemic diets led to a significant increase of oxidative stress as shown by GSH depletion in aorta and liver, and an increased MDA level in liver and heart in agreement with previous studies.16,20,65,66 Metabolomics analysis also revealed other metabolic changes induced by the high-fat diet and in particular an increase in the urinary excretion of deoxycytidine, nicotinic acid, 2,4-dihydroxyquinoline and pipecolinic acid (Figure 4). A high urine excretion of deoxycytidine nucleosides has been associated to a higher risk of cancer and immune deficiency syndrome.67,68 An increase of deoxycytidine excretion in urine with high-fat diets has not been reported before. It might be due to an increase in DNA breakdown or to a higher production of deoxycytidine by enteric bacteria.68 2,4-Dihydroxyquinoline has been recently shown to be produced by the bacteria Pseudomonas aeruginosa, a human pathogen, from anthranilic acid, a tryptophan metabolite.69 The origin of the variations in the urinary excretion of dihydroxyquinoline observed in the present study following high-fat diets remains unclear. Concerning pipecolinic acid, its high urinary content generally results from either chronic liver dysfunction70 or peroxisomal disorders.71,72 Indeed, peroxisome abnormalities lead to impaired pipecolinic acid oxidation and to its accumulation in biofluids such as urine and plasma.71,72 Increased pipecolinic acid excretion following high-fat diets has never been reported before. One may only speculate that hyperlipidic diets lead to some liver dysfunction associated to peroxisomal disorders. Catechin supplementation had no effect on the oxidative stress induced by the high-fat diet except on urine MDA. Previous results showed contrasted effects of catechin on antioxidant status: for example, catechin-rich food (tea, wine and cocoa) consumption was shown in humans to increase plasma antioxidant capacity and decrease urinary F2-isoprostanes, while other studies showed no effect.57 Plasma choles-
481.1351
467.1194
6.9
7.7
467.1195
7.4
481.1358
291.0883
7.8
8.3
196.0630
6.3
305.1032
180.0680
7.0
8.6
detected mass m/z [M+H]+
RT
134.0613 106.0402 105.0348 218.0470 122.0336 121.0305 292.0930 273.0782 165.0593 139.0404 123.0458 468.1230 449.1082 431.1003 291.0883 292.0921 273.0820 165.0588 139.0405 123.0469 505.0788 484.1455 468.1223 431.1013 291.0875 292.0932 273.0795 165.0600 139.0406 179.0742 147.0468 139.0401 137.0638 961.2650 498.1629 482.1400 463.1238 445.1149 306.1073 305.1033 288.0974 287.0939 179.0734 139.0411 137.0619 482.1367 305.1026 287.0940
[M+H-carboxyl] (13C isotope [M+H-glycine]+) [M+H-glycine]+ [M+Na]+ (13C isotope [M+H-glycine]+) [M+H-glycine]+ (13C isotope [M+H]+) [M+H-H2O]+ 1,4 + B 1,3 + A 1,2 + B (13C isotope [M+H]+) [M+H-H2O]+ [M+H-2H2O]+ [M+H-glucuronide]+ (13C isotope [M+H]+) [M+H-H2O]+ 1,4 + B 1,3 + A 1,2 + B [M+K]+ [M+NH4]+ (13C isotope [M+H]+) [M+H-2H2O]+ [M+H-glucuronide]+ (13C isotope [M+H]+) [M+H-H2O]+ 1,4 + B 1,3 + A 1,4 + B [1,4B+-H2O] 1,3 + A 1,2 + B [2M-H]+ [M+NH4]+ (13C isotope [M+H]+) [M+H-H2O]+ [M+H-2H2O]+ (13C isotope [M+H-glucuronide]+) [M+H-glucuronide]+ (13C isotope [M+H-glucuronide-H2O]+) [M+H-glucuronide-H2O]+ 1,4 + B 1,3 + A 1,2 + B (13C isotope [M+H]+) [M+H-glucuronide]+ [M+H-glucuronide-H2O]+
+
positive ESI MS fragmentsa
Catechin glucuronide
467.1183
Methylcatechin glucuronide
Methylcatechin glucuronide
481.1340
Methylcatechin
481.1340
305.1019
Catechin glucuronide
Catechinb
291.0863
467.1183
Hydroxyhippuric acidb
Hippuric acidb
putative identification
196.0604
180.0655
calculated mass m/z [M+H]+
Table 4. Catechin Metabolites (derivatives and microbial) and Their Fragments and Adducts Formed in the Electrospray Ionization Source
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385.1129
223.0992
399.1313
303.0557
7.7
9.1
8.0
9.7
137.0617 445.1375 423.0642 407.0990 403.1448 402.1428 367.1046 210.0879 209.0859 191.0473 205.0927 164.0718 163.0786 437.0874 421.1117 417.1606 416.1582 381.1186 224.1031 223.0987 205.0892 163.0786 320.0826 285.1685 240.1264 223.0972 163.0786 B [M+K+Na]+ [M+K]+ [M+Na]+ (13C isotope [M+NH4]+) [M+NH4]+ [M+H-H2O]+ (13C isotope [M+H-glucuronide]+) [M+H-glucuronide]+ [M+H-glucuronide-H2O]+ [M+H-H2O]+ (13C isotope [M-C2H3O2]+) [M-C2H3O2]+ [M+K]+ [M+Na]+ (13C isotope [M+K]+) [M+K]+ [M+H- H2O]+ (13C isotope [M+H-glucuronide]+) [M+H-glucuronide]+ [M+H-glucuronide-H2O]+ [M-C2H3O2]+ [M+NH4]+ [M+H-H2O]+ [M+NH4-sulfate]+ [M+H-sulfate]+ [M-C2H3O2]+
+
positive ESI MS fragmentsa 1,2
303.0533
399.1285
223.0964
385.1129
calculated mass m/z [M+H]+
Methoxy-hydroxyphenylvalerolactone sullfate
Methoxy-hydroxyphenylvalerolactone glucuronide
Methoxy-hydroxyphenylvalerolactone
Dihydroxyphenylvalerolactone glucuronide
putative identification
a 1,4 + B and 1,2B+ are fragments of catechin and of its metabolites as described by Cren-Olive´ et al.56 For catechin glucuronide, the fragments 1,3A+, 1,4B+ and 1,2B+ corresponded to those of [M-glucuronide]+. The 14 Da shift of both methylcatechin and methylcatechin glucuronide fragments (1,2B+ ) 137 ) 123 + 14 and 1,4B+ ) 179 ) 165 + 14) corresponds to the methyl moiety. b Identity confirmed with authentic standards.
detected mass m/z [M+H]+
RT
Table 4. Continued
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Metabolic Effects of Catechin in Rats Fed High-Fat Diets
Figure 5. Evolution of the spectral intensity of nicotinic acid (m/z 124.041) in urine of rats after feeding low-fat (LF) and mediumfat (MF) supplemented or not with catechin (PP), at day 38. NS, not significant. Values are means ( SEM (n ) 6 rats/group).
terol and TG levels were not modified upon catechin supplementation. As for the antioxidant status, contradictory results have been published; some authors reported hypocholesterolemic effects of catechin in rats or rabbits, but at a dose of catechin 10 times higher than the one used here.73,74 On the other hand, catechin supplementation had a pronounced effect on the urinary metabolome as revealed by the fingerprinting approach used in the present study. These changes are partly explained by the excretion of catechin and catechin metabolites. Several of these metabolites could be identified. Most were known metabolites of catechin such as O-methyl catechin, catechin glucuronides, methylcatechin glucuronide, hydroxyphenylvalerolactone metabolites, hippuric acid and hydroxyhippuric acid formed in the tissues or by the microbiota in the colon.50,51,75–79 The presence of methoxyhydroxyphenylvalerolactone glucuronide, not previously described as a catechin metabolite, was also suggested based on its MS spectrum. Other urinary metabolites affected by catechin supplementation were endogenous metabolites reflecting the impact of the catechin on the rat metabolism. Catechin was shown in particular to reverse some of the metabolic changes induced by the high-fat diets. These effects appear substantial as shown by the large number of reversed variables. One of these variables is nicotinic acid which increased with the content of fat in the diet and decreased upon catechin supplementation (Figure 5). Nicotinic acid is a water-soluble vitamin which plays an essential role in energy metabolism and DNA repair. The increased urinary excretion of nicotinic acid with the high-fat diets may reflect an increased conversion rate of tryptophan into nicotinic acid. A high-fat intake is known to increase the conversion rate of tryptophan to nicotinamide, a precursor of nicotinic acid.80–82 These effects have been explained by a down-regulation of R-amino-β-carboxymuconate--semialdehyde decarboxylase (ACMSD), a key enzyme diverting the conversion of tryptophan toward R-aminomuconate--semialdehyde and glycolysis.81,82 Peroxisome proliferator-activated receptor R (PPARR) might be involved in these effects as its activation is known to inhibit ACMSD and to play a critical role in the control of the tryptophan-nicotinic acid pathway.83 Catechin might also influence the regulation of this pathway through some interactions with PPARR. Some polyphenols such as isoflavones and resveratrol showed some agonist activities rather than antagonist effects of PPARR, but antagonist effects were also observed depending on time factors.84,85 These effects
merit to be further explored. Nicotinic acid is also known to be synthesized by bacteria.86 The decrease in excretion of nicotinic acid in urine by catechin might also indirectly result from some modification of the microbiota profile induced by catechin supplementation.87 These metabolic effects of catechin revealed by metabolomics still need to be confirmed in future studies. They may help to understand the hypolipemic and antiobesity effects of tea catechins previously described.62,88 The full elucidation of the metabolic effects of catechin will require a more complete identification of the markers of effects detected in the present experiment. More efficient softwares or more comprehensive databases will be needed to identify these markers and develop a full biological pathway interpretation. We may hope that it will allow to identify the key metabolic pathways affected by the intake of polyphenols or other antioxidants, beyond their sole antioxidant effects. Abbreviations: ACMSD, R-aminomuconate--semialdehyde; FRAP, ferring reducing ability of plasma; GSH, reduced glutathione; GSSG, oxidized glutathione; HF, high fat; LC-QTOF, liquid chromatography-quadrupole time-of-flight; LF, low fat; MDA, malondialdehyde; MF, medium fat; MS, mass spectrometry; PCA, principal component analysis; PLS-DA, partial least-square discriminant analysis; PPAR, peroxisome proliferator-activated receptor; SOD, superoxide dismutase; TBARS, thiobarbituric acid reactive substances.
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PR800034H
