Premium
In vitro oxidation of vitamins C and E, cholesterol, and thiols in rat brain synaptosomes
Author(s) -
Vatassery G. T.
Publication year - 1995
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02536285
Subject(s) - chemistry , vitamin e , antioxidant , chromatography , biochemistry , oxidative phosphorylation , cholesterol , saponification , tocopherol , lipid peroxidation , clinical chemistry
Free radical‐induced oxidation of vitamins C, E, sulfhydryl compounds, and cholesterol in brain synaptosomes from Fisher 344 rats was studied. The synaptosomes were incubated at 37°C with 2,2′‐abo bis ‐(2‐amidinopropane) dihydrochloride (AAPH), which undergoes thermal decomposition to yield free radicals. After incubation, the synaptosomes were sedimented, saponified, and extracted with hexane to isolate tocopherol and cholesterol. Ascorbate and tocopherol were assayed by liquid chromatography, cholesterol by gas chromatography, and total sulfhydryls by spectrophotometry. Under the in vitro conditions used in this study, the approximate order for the ease of oxidation of the various compounds was: ascorbate ≫tocopherol>sulfhydryl compounds⋙cholesterol. However, tocopherol and sulfhydryl oxidation occurred even before all of the ascorbate had been consumed. Therefore, the fate of a specific antioxidant at a particular cellular location cannot be predicted with complete accuracy using the in vitro order for ease of oxidation shown here. Ascorbate may play a major role in protecting brain against oxidative damage because: (i) ascorbate concentration is high in brain, (ii) it can regenerate vitamin E from its radical oxidation product, and (iii) it is one of the first antioxidants to be consumed during oxidative reactions.