Vitamin E sensitive genes in the developing rat fetal brain: A high-density oligonucleotide microarray analysis

Roy S, Lado BH, Khanna S, Sen CK.

FEBS Lett. 2002 Oct 23;530(1-3):17-23.

Vitamin E (tocopherols and tocotrienols) is essential for normal neurological function. Recently we have reported that the neuroprotective properties oftocotrienols are much more potent than that of the widely studied tocopherols (Sen, C.K., Khanna, S., Roy, S. and Parker, L. (2000) J. Biol. Chem. 275, 13049-13055). The objective of this study was to evaluate whether (i) oral supplementation of tocotrienols during pregnancy is bioavailable to fetal and mother brains; (ii) short-term change in dietary vitamin E levels of pregnant rats influences gene expression profile of developing fetal brains. We report that dietary tocotrienol is bioavailable to both mother and fetal brains. The enrichment is more in fetal brain tissue. Using a GeneChip microarray expression profiling approach we have identified a specific set of vitamin E sensitive genes in the developing rat fetal brain.

Vitamin C, tocopherols, and tocotrienols in berries of wild and cultivated sea buckthorn (Hippophaë rhamnoides L.) of different origins and harvesting dates were determined with HPLC. Wild berries of subsp. sinensis, native to China, contained 5-10 times more vitamin C in the juice fraction than the berries of subsp. rhamnoides from Europe and subsp. mongolica from Russia (4-13 vs 0.02-2 g/L juice). Genetic background and berry-harvesting date were two primary factors determining the vitamin C content in the berries. Crossing different subspecies influenced the vitamin C content to some extent. For bushes cultivated in southwest Finland, the best berry-harvesting date for high vitamin C content was the end of August. The seeds of subsp. sinensis contained less tocopherols and tocotrienols (average 130 mg/kg) compared with seeds of subsp. rhamnoides (average 290 mg/kg) and mongolica (average 250 mg/kg). The fruit flesh of sinensis berries had contents of tocopherols and tocotrienols 2-3 times higher than those found in the other two subspecies (120 mg/kg vs 40 mg/kg in rhamnoides and 50 mg/kg in mongolica). The fresh whole berries of subsp. sinensis were clearly the best source of total tocopherols and tocotrienols. The total content of tocopherols and tocotrienols in the soft parts of the berries reached the maximum level around early- to mid-September, whereas the content in seeds continued to increase until the end of November. The excellent combination of the highest content of vitamin C and tocopherols and tocotrienols makes the berries of subsp. sinensis an optimal raw material for nutritional investigation as a candidate for functional foods with special antioxidative properties.

We have previously shown that alpha-tocotrienol (alpha-T3), a vitamin E analogue and HMG CoA reductase (HMGR) inhibitor, markedly inhibited monocyte-endothelial cell adhesion, a process that was reversed with the addition of mevalonate intermediates involved in protein prenylation. Since delta-T3 and gamma-T3 possess greater HMGR inhibition than alpha-T3, we postulated that these analogues might have a greater effect on protein prenylation, and thus on monocyte adhesion and endothelial adhesion molecule expression in comparison to alpha-T3. Hence, we pursued to investigate the effect of various analogues of tocotrienol (alpha, gamma, delta) on monocytic cell adhesion and expression of adhesion molecules using a human umbilical vein endothelial cell-line, EA.hy926, as the model system. Relative to alpha-T3, delta-T3 displayed a more profound inhibitory effect on monocytic cell adherence using a 15 micromol/L concentration within 24 h (delta: 42 +/- 5%; alpha: 26 +/- 8% vs. control). This inhibitory action was reversed by co-incubation with farnesol and geranylgeraniol, suggesting a role for prenylated proteins in the regulation of monocyte adhesion. To further evaluate the effect of tocotrienols on the vascular endothelium, we measured the surface expression of adhesion molecules. Compared to alpha-T3, delta-T3 markedly inhibited the expression of VCAM-1 (delta: 57 +/- 6%; alpha: 37 +/- 10% vs. control) and E-selection (delta: 36 +/- 3%; alpha: 18 +/- 6% vs. control) in TNF-alpha activated endothelial cells. The above result suggests that delta-T3 is a potent and effective agent for the reduction of cellular adhesion molecule expression and monocytic cell adherence.

Identities and differences in the metabolism of tocotrienols and tocopherols in HepG2 cells

Birringer M, Pfluger P, Kluth D, Landes N, Brigelius-Flohé R.

J Nutr. 2002 Oct;132(10):3113-8.

The metabolism of alpha- and gamma-tocotrienol was investigated in HepG2 cells. Metabolites were identified by HPLC and gas chromatography/mass spectrometry. gamma-Tocotrienol was degraded to gamma-CEHC (carboxyethyl hydroxychroman), gamma-CMBHC (carboxymethylbutyl hydroxychroman), gamma-CMHenHC (carboxymethylhexenyl hydroxychroman), gamma-CDMOenHC (carboxydimethyloctenyl hydroxychroman) and gamma-CDMD(en)(2)HC (carboxydimethyldecadienyl hydroxychroman). alpha-Tocotrienol yielded alpha-CEHC, alpha-CMBHC, alpha-CMHenHC and alpha-CDMOenHC, whereas alpha-CDMD(en)(2)HC could not be detected. These findings demonstrate that the trienols are metabolized essentially like tocopherols, i.e., by omega-oxidation followed by beta-oxidation of the side chain. The failure to detect CMBHC with the original double bond in the side chain reveals that auxiliary enzymes are involved, as in the metabolism of unsaturated fatty acids. CMBHC were the most abundant metabolites obtained from the tocotrienols as well as from alpha-tocopherol. Quantitatively, the tocotrienols were degraded to a larger extent than their counterparts with saturated side chains. The pronounced quantitative differences in the metabolism between individual tocopherols as well as between tocotrienols and tocopherols in vitro suggest a corresponding lack of equivalence in vivo.

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