The synthesis of vitamin C is substantially reduced in Osteogenic Disorder Shionogi (ODS) rats. Hepatocytes prepared from these rats contained ~12% of the wild-type content of this vitamin. In culture, the ascorbate content remained low in the absence of supplementation of the medium. Independent of their vitamin C status, cultured hepatocytes become depleted of vitamin E. Supplementation of the culture medium with 100 μM ascorbate and 1.2 μM α-tocopherol phosphate maintained the physiological content of both vitamins C and E in ODS hepatocytes. Thus, the antioxidant function of vitamins C and E could be evaluated in the presence of both or either vitamin or in the absence of both vitamins. Hepatocytes deficient in both vitamins were the most susceptible to lipid peroxidation (as measured by thiobarbituric acid) and to cell killing within a 90-min exposure to 125-500 μM tert-butyl hydroperoxide (TBHP). Supplementation to achieve a physiological content of both vitamins C and E reduced the evidence of lipid peroxidation and abolished the cell killing. Supplementation with either vitamin alone resulted in an intermediate degree of both lipid peroxidation and cell killing. In ODS hepatocytes treated with TBHP, the decline in vitamin E preceded the decline in vitamin C. In ODS hepatocytes depleted of vitamin C, the loss of vitamin E after exposure to TBHP was greater than that in the presence of physiological levels of ascorbate. This greater loss of vitamin E in the face of a depletion of vitamin C was readily attributable to the increased peroxidation of lipids. Thus, the physiological level of vitamin C in cells does not seem to regenerate vitamin E. In contrast, the rate and extent of the depletion of vitamin C correlate with the degree of cell killing. These data document the antioxidant function of the physiological level of cellular vitamin C and relate this function to protection against peroxidative cell injury.
|Original language||English (US)|
|Number of pages||6|
|State||Published - Oct 1996|
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