Gamma-tocopherol Metabolism Research Articles

Metabolism of Gamma-tocopherol in the Liver of Male Sprague-Dawley Rat

Vitamin E (VE) plays important roles in the physiological and biochemical processes of animals and humans. However, VE is an unique vitamin that has not been understood thoroughly because the metabolites of VE are very complicated and hard to be analysed. This experiment was conducted to study the metabolism of γ-tocopherol, one of 8 VE isomers, in the liver post-mitochondrial supernatant (S9) of male Sprague-dawley rat (Rattus norvegiens, SD-rat) using a novel approach. The results showed that an optimal reactive system of γ-tocopherol metabolism in vitro was composed of 5 mg/ml of S9, 8 μg/ml of γ-tocopherol, 1 mmol/ml of nicotine amide adenine dinucleotide phosphate (NADPH) and 45 min of reactive time at 35°C. The metabolites of γ-tocopherol could be successfully analyzed using gradient elution and reverse-phase high-performance liquid chromatography (RP-HPLC). A main metabolite was identified as ω-hydroxylate, a side chain of γ-tocopherol, when the structure of γ-tocopherol metabolites was determined using gas chromatography-mass spectrometry (GC-MS). A conclusion was drawn that ω-hydroxylate, a main metabolite of γ-tocopherol, could be most utilized by SD-rat and other mammals.

Effects of alpha-tocopherol and sesamin on gamma-tocopherol metabolism in rats

Effects of alpha-tocopherol and sesamin on gamma-tocopherol metabolism in rats

Cytochrome P450‐Dependent Metabolism of Vitamin E Isoforms is a Critical Determinant of Their Tissue Concentrations in Rats

The aim of this study was to clarify the contribution of cytochrome P450 (CYP)-dependent metabolism of vitamin E isoforms to their tissue concentrations. We studied the effect of ketoconazole, a potent inhibitor of CYP-dependent vitamin E metabolism in cultured cells, on vitamin E concentration in rats. Vitamin E-deficient rats fed a vitamin E-free diet for 4 weeks were administered by oral gavage a vitamin E-free emulsion, an emulsion containing alpha-tocopherol, gamma-tocopherol or a tocotrienol mixture with or without ketoconazole. Alpha-tocopherol was detected in the serum and various tissues of the vitamin E-deficient rats, but gamma-tocopherol, alpha- and gamma-tocotrienol were not detected. Ketoconazole decreased urinary excretion of 2,5,7,8-tetramethyl-2(2'-carboxyethyl)-6-hydroxychroman after alpha-tocopherol or a tocotrienol mixture administration, and that of 2,7,8-trimethyl-2(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC) after gamma-tocopherol or a tocotrienol mixture administration. The gamma-tocopherol, alpha- and gamma-tocotrienol concentrations in the serum and various tissues at 24 h after their administration were elevated by ketoconazole, while the alpha-tocopherol concentration was not affected. The gamma-tocopherol or gamma-tocotrienol concentration in the jejunum at 3 h after each administration was also elevated by ketoconazole. In addition, significant amount of gamma-CEHC was in the jejunum at 3 h after gamma-tocopherol or gamma-tocotrienol administration, and ketoconazole inhibited gamma-tocopherol metabolism to gamma-CEHC in the jejunum. These results showed that CYP-dependent metabolism of gamma-tocopherol and tocotrienol is a critical determinant of their concentrations in the serum and tissues. The data also suggest that some amount of dietary vitamin E isoform is metabolized by a CYP-mediated pathway in the intestine during absorption.

Dietary Sesame Seed Decreases Urinary Excretion of .ALPHA.- and .GAMMA.-Tocopherol Metabolites in Rats

We previously showed that dietary sesame seed and its lignan inhibited gamma-tocopherol metabolism to 2,7,8-trimethyl-2(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), a gamma-tocopherol metabolite, and markedly elevated tissue gamma-tocopherol concentration in rats. The aim of this study was to clarify the effect of dietary sesame seed on alpha-tocopherol metabolism. Vitamin E-deficient rats fed a vitamin E-free diet for 4 wk were fed a diet containing alpha-tocopherol, alpha- and gamma-tocopherol, or alpha-tocopherol with sesame seed for 7 d. Urinary excretion of 2,5,7,8-tetramethyl-2(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC), a alpha-tocopherol metabolite, in rats fed alpha-tocopherol with sesame seed was inhibited (p<0.05) as compared with that in rats fed alpha-tocopherol alone, or alpha- and gamma-tocopherol. The gamma-CEHC excretion was also less (p<0.05) in rats fed alpha-tocopherol with sesame seed than that in rats fed alpha- and gamma-tocopherol. The inhibition of alpha- and gamma-CEHC excretion by sesame seed was accompanied by elevation (p<0.05) of the alpha- and gamma-tocopherol concentration in the liver. These results suggest that dietary sesame seed inhibits not only gamma-tocopherol metabolism to gamma-CEHC but also alpha-tocopherol metabolism to alpha-CEHC in rats.

Alpha- and gamma-tocopherol in women with polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is associated with a broad range of metabolic and endocrine manifestations including increased oxidative stress, systemic inflammation and insulin resistance. Alpha-tccopherol (AT) and gamma-tocopherol (GT) play an important role in protection from oxidative stress by scavenging reactive oxygen and nitrogen oxide species. Recent studies suggest that the properties and metabolism of GT are distinctly different from AT. Specifically, GT has a particular affinity to the nitrogen oxide species and possesses unique anti-inflammatory properties related to inhibition of cyclooxygenase activity.

Cereal Alkylresorcinols Elevate γ-Tocopherol Levels in Rats and Inhibit γ-Tocopherol Metabolism In Vitro

Alkylresorcinols (AR) are a class of amphiphilic phenolic lipids present in high amounts in wheat and rye bran. They have been reported to be both growth retarding and innocuous when fed to rats, and to have a broad range of bioactivities in vitro, suggested to be related to their ability to bind to proteins and modify membranes. This study was designed to test the effects of AR (purified from rye bran) on growth, tocopherol levels, and cholesterol levels in rats. Rats were fed 1 of 4 different levels of AR for 4 wk: 0 (control), 1, 2, and 4 g/kg diet. AR did not affect final body, liver, or lung weights. The AR diets increased the levels of gamma-tocopherol in liver and lungs (P < 0.05). To investigate whether AR could have increased gamma-tocopherol levels via inhibition of tocopherol-omega-hydroxylase, HepG2 cells were incubated with AR and the metabolism of gamma-tocopherol measured. AR significantly inhibited the conversion of gamma-tocopherol to its water-soluble hydroxychroman metabolite in vitro, indicating that AR may increase gamma-tocopherol levels via inhibition of tocopherol metabolism in vivo. The 4 g AR/kg diet decreased liver cholesterol (P < 0.001), but did not affect plasma lipids. AR were detected in the perirenal adipose tissue samples of rats fed AR, indicating that they can accumulate in the fatty tissues of rats. High levels of dietary AR moderately affect gamma-tocopherol, possibly via inhibition of tocopherol metabolism, and decrease liver cholesterol in rats.

Alpha and gamma tocopherol metabolism in healthy subjects and patients with end-stage renal disease

The metabolism of alpha and gamma tocopherol, the major components of vitamin E, have not been studied in uremic patients. The major pathway of tocopherol metabolism is via phytyl side chain oxidation, leaving carboxyethyl-hydroxychromans (CEHC) as metabolites. Alpha and gamma CEHC are water soluble, renally excreted, with known potent anti-inflammatory and antioxidative properties. We examined serum alpha and gamma tocopherol and respective CEHC concentrations in 15 healthy subjects and 15 chronic hemodialysis patients. Serum alpha tocopherol levels were similar in hemodialysis patients (12.03 +/- 1.34 microg/mL) and healthy subjects (11.21 +/- 0.20 microg/mL), while serum gamma tocopherol levels were significantly greater in hemodialysis patients (3.17 +/- 0.37 microg/mL) compared to healthy subjects (1.08 +/- 0.06 microg/mL, P < 0.0001). Serum alpha and gamma CEHC levels were tenfold and sixfold higher in hemodialysis patients compared to healthy subjects, respectively (both P < 0.0001). Serum alpha and gamma tocopherol levels and CEHC metabolites were also measured after supplementation of alpha- or gamma-enriched mixed tocopherols in both hemodialysis patients and healthy subjects. Tocopherol administration resulted in modest or nonsignificant changes in serum tocopherol concentrations, while markedly increasing serum CEHC concentrations in both healthy subjects and hemodialysis patients. Hemodialysis resulted in no change in the serum alpha or gamma tocopherol concentrations while decreasing serum alpha CEHC and gamma CEHC levels by 63% and 53%, respectively (both P = 0.001 versus predialysis). Fourteen-day administration of gamma-enriched but not alpha tocopherols lowered median C-reactive protein (CRP) significantly in hemodialysis patients (4.4 to 2.1 mg/L, P < 0.02). First, serum alpha and gamma CEHC accumulate in uremic patients compared to healthy subjects; second, supplementation with tocopherols dramatically increases serum CEHC levels in both healthy subjects and hemodialysis patients; and, finally, CEHC accumulation may mediate anti-inflammatory and antioxidative effects of tocopherols in hemodialysis patients.

Measurement of vitamin E metabolites by high-performance liquid chromatography during high-dose administration of alpha-tocopherol.

alpha-Tocopherol and gamma-tocopherol are metabolized into 2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC) and 2,7,8-trimethyl-2-(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), respectively. We analyzed alpha- and gamma-CEHC concentrations in human serum and urine by high-performance liquid chromatography during administration of alpha-tocopherol. Fourteen healthy adult male volunteers received 1,200 IU per day of RRR-alpha-tocopherol for 28 days. Blood and urine samples were obtained on days 0, 14, 28, and 56. During alpha-tocopherol administration, the plasma gamma-tocopherol concentration decreased significantly, but there was marked elevation of the alpha-tocopherol concentration. Increased concentration of alpha-CEHC and gamma-CEHC in both serum and urine indicated the acceleration of vitamin E metabolism. High-dose administration of alpha-tocopherol caused an increase of gamma-tocopherol metabolism, which might have caused a decrease of the plasma gamma-tocopherol concentration.

Cytochrome P450 ω-Hydroxylase Pathway of Tocopherol Catabolism: NOVEL MECHANISM OF REGULATION OF VITAMIN E STATUS

Postabsorptive elimination of the various forms of vitamin E appears to play a key role in regulation of tissue tocopherol concentrations, but mechanisms of tocopherol metabolism have not been elucidated. Here we describe a pathway involving cytochrome P450-mediated omega-hydroxylation of the tocopherol phytyl side chain followed by stepwise removal of two- or three-carbon moieties, ultimately yielding the 3'-carboxychromanol metabolite that is excreted in urine. All key intermediates of gamma-tocopherol metabolism via this pathway were identified in hepatocyte cultures using gas chromatography-mass spectrometry. NADPH-dependent synthesis of the initial gamma- and alpha-tocopherol 13'-hydroxy and -carboxy metabolites was demonstrated in rat and human liver microsomes. Functional analysis of several recombinant human liver P450 enzymes revealed that tocopherol-omega-hydroxylase activity was associated only with CYP4F2, which also catalyzes omega-hydroxylation of leukotriene B(4) and arachidonic acid. Tocopherol-omega-hydroxylase exhibited similar binding affinities but markedly higher catalytic activities for gamma-tocopherol than alpha-tocopherol, suggesting a role for this pathway in the preferential physiological retention of alpha-tocopherol and elimination of gamma-tocopherol. Sesamin potently inhibited tocopherol-omega-hydroxylase activity exhibited by CYP4F2 and rat or human liver microsomes. Since dietary sesamin also results in elevated tocopherol levels in vivo, this pathway appears to represent a functionally significant means of regulating vitamin E status.

Dietary Sesame Seed and Its Lignans Inhibit 2,7,8-Trimethyl- 2(2′-carboxyethyl)-6-hydroxychroman Excretion into Urine of Rats Fed γ-Tocopherol

We showed previously that dietary sesame seed and its lignans elevate the tocopherol concentration in rats. To clarify their effect on tocopherol metabolism, we determined in this study the urinary excretion of 2,7,8-trimethyl-2(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), a gamma-tocopherol metabolite, in rats fed sesame seed or its lignans. Rats were fed diets with or without sesame seed for 28 d in Experiment 1, and for 1, 3 and 7 d in Experiment 2. On d 28, dietary sesame seed elevated (P < 0.05) gamma-tocopherol concentrations in liver, kidney, brain and serum, and decreased (P < 0.05) urinary excretion of gamma-CEHC. The excretion was completely inhibited by feeding sesame seed on d 1 and 3. In Experiment 3, the effects of dietary sesamin and sesaminol (major lignans in sesame seed) or ketoconazole (a selective inhibitor of cytochrome P(450) (CYP)3A on urinary excretion of gamma-CEHC in rats fed gamma-tocopherol were examined. The urinary gamma-CEHC in rats fed sesamin or sesaminol was markedly lower than in rats fed gamma-tocopherol alone (P < 0.05). Dietary ketoconazole also inhibited (P < 0.05) urinary excretion of gamma-CEHC, and elevated (P < 0.05) gamma-tocopherol concentrations in tissues and serum of rats fed gamma-tocopherol. These data suggest that sesame seed and its lignans elevate gamma-tocopherol concentration due to the inhibition of CYP3A-dependent metabolism of gamma-tocopherol.