Comparison of RRR-alpha- and all-rac-alpha-tocopherol uptake by permanent rat skeletal muscle myoblasts (L6 cells): effects of exogenous lipoprotein lipase.
The purpose of the present investigation was to test whether permanent skeletal muscle cells (rat L6 cells) could serve as an in vitro model for alpha-tocopherol (alphaTocH) biodiscrimination studies. L6 cells were incubated in the presence of high density lipoprotein (HDL), low density lipoprotein (LDL), and very low density lipoprotein (VLDL) labeled in the lipid moiety with either all-rac- or RRR-[14C]alphaTocH. These incubations were performed either in the absence or in the presence of exogenously added bovine lipoprotein lipase (LPL) since skeletal muscle is one of the major expression sites of LPL in vivo. Time-dependent uptake studies (up to 24 h) in the absence of LPL have shown that equipotent doses of all-rac- and RRR-[14C]alphaTocH (1.36:1) led to almost identical accumulation of the tracer, independent of the lipoprotein class used as alphaTocH carrier. With regard to alphaTocH donor capacity, it appeared that HDL is the most potent alphaTocH donor, followed by LDL and VLDL. In the presence of LPL, all-rac- and RRR-[14C]alphaTocH uptake was significantly enhanced (between two- and tenfold). Biodiscrimination studies using chiral high-performance liquid chromatographic analysis with radiometric detection of the corresponding methyl ether derivatives on a Chiralcel OD column have demonstrated that the 2S-and 2R-isomers of alphaTocH were taken up in a 1:1 ratio by L6 cells independent of the absence or presence of LPL. In addition, we have not observed biodiscrimination between the four 2R-isomers, i.e., there was no preferential accumulation of the RRR-isomer. These data suggest that L6 cells do not discriminate between different alphaTocH isomers and that the addition of endogenous LPL significantly enhances the uptake of RRR- and all-rac-alphaTocH.
Nakamura
T
T
Eisai Co., Ltd., Vitamin E Technology Section, Tokyo, Japan.
Reicher
H
H
Sattler
W
W
D000818:Animals; D002250:Carbon Radioisotopes; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007536:Isomerism; D008071:Lipoprotein Lipase / Q000494:pharmacology*; D008075:Lipoproteins, HDL / Q000378:metabolism / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism / Q000494:pharmacology; D008079:Lipoproteins, VLDL / Q000378:metabolism / Q000494:pharmacology; D018482:Muscle, Skeletal / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011874:Radiometry; D051381:Rats; D024505:Tocopherols; D014810:Vitamin E / Q000031:analogs & derivatives / Q000737:chemistry* / Q000493:pharmacokinetics*; D024502:alpha-Tocopherol / Q000031:analogs & derivatives*
D002250:Carbon Radioisotopes; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D014810:Vitamin E; D008071:Lipoprotein Lipase; D024502:alpha-Tocopherol; D024505:Tocopherols