Abstract
The effect of pretreatment with ascorbic acid (vitamin C) on chromate-induced DNA damage, cytotoxicity, and enzyme inhibition as well as on the cellular reduction of chromium(VI) was investigated using Chinese hamster V-79 cells. Cellular pretreatment with nontoxic levels of 1 mM ascorbic acid for 24 h prior to exposure resulted in a significant increase (1.7-fold) in cellular levels of this vitamin. Alkaline elution assays demonstrated that this pretreatment decreased cellular levels of Na2CrO4-induced alkali-labile sites while the numbers of DNA-protein crosslinks produced by chromate increased. In colony-forming assays, pretreatment with ascorbic acid enhanced the cytotoxicity of chromate. However, the inhibition of glutathione reductase attributed to Na2CrO4 was attenuated by this pretreatment. Under the same experimental condition, the uptake of chromate in pretreated cells was found to increase. ESR studies revealed that cellular pretreatment with ascorbic acid reduced the level of chromium(V) intermediate and increased the level of chromium(III) complex, indicating that cellular reduction of chromium(VI) to chromium(III) was accelerated by this vitamin. These results suggest that ascorbic acid decreases chromate-induced alkali-labile sites and chromium inhibition of glutathione reductase, but it enhances DNA-protein cross-links and cytotoxicity caused by this metal through its ability to directly reduce chromium(VI).
Highlights
ESR studies revealed that cellular pretreatment with ascorbic acid reduced the level ofchromium(V) intermediate and increased the level of chromium(II1)complex, indicating that cellular reduction of chromium(V1) to chromodification of cellular levels of glutathione, cytochrome P450 reductase, andriboflavin was found toaffect the levels of DNA damage produced by chromate compounds in cultured cells (13,15)
We examined the effect of pretreatment with ascorbic acid on chromate-induced DNA damage, enzyme inhibition, andcytotoxicity in Chinese hamster V-79 (V-79)’ cells
We used this concentration through- both the alkali-labile sites and the enzyme inhibitioncaused out the subsequent experiments
Summary
ESR studies revealed that cellular pretreatment with ascorbic acid reduced the level ofchromium(V) intermediate and increased the level of chromium(II1)complex, indicating that cellular reduction of chromium(V1) to chromodification of cellular levels of glutathione, cytochrome P450 reductase, andriboflavin was found toaffect the levels of DNA damage produced by chromate compounds in cultured cells (13,15). We examined the effect of pretreatment with ascorbic acid on chromate-induced DNA damage, enzyme inhibition, andcytotoxicity in Chinese hamster V-79 (V-79)’ cells.
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