Difference in uptake and toxicity of trivalent and pentavalent inorganic arsenic in rat heart microvessel endothelial cells

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Abstract
Intake of inorganic arsenic is known to cause vascular diseases as well as skin lesions and cancer in humans. We investigated the differences in cytotoxicity, uptake rate of arsenic, and gene expression of antioxidative enzymes between arsenite (As3+)- and arsenate (As5+)-exposed rat heart microvessel endothelial cells. As3+ was more cytotoxic than As5+, and LC50 values were calculated to be 36 and 220 µM, respectively. As3+ (1–25 µM) increased mRNA levels of antioxidant enzymes such as heme oxygenase-1 (HO-1), thioredoxin peroxidase 2, NADPH dehydrogenase, and glutathione S-transferase P subunit. HO-1 mRNA levels showed the most remarkable increase in response to As3+. cDNA microarray analysis indicated that there was no prominent difference in arsenic-induced transcriptional changes between As3+- and As5+-exposed cells, when the cells were exposed to one-fourth the LC50 concentration of arsenic (9 and 55 µM for As3+ and As5+, respectively). N-acetyl-l-cysteine (NAC) reduced both the cytotoxicity of inorganic arsenic and the HO-1 mRNA level, and buthionine sulfoximine enhanced cytotoxicity of inorganic arsenic. As3+ was taken up by the endothelial cells 6–7 times faster than As5+, and the presence of NAC in the culture medium did not change the uptake rate of As3+.These results suggest that the effects of NAC on arsenic-induced cytotoxicity and oxidative stress were due to the antioxidative role of non-protein thiols and not to chelation of arsenic in the culture medium. The difference in cellular uptake of arsenic between As3+ and As5+ appeared not to be due to the ionic charge on arsenic (at physiological pH, trivalent arsenic is neutral whereas pentavalent arsenic is negatively charged). These results suggest that the higher toxicity of As3+ compared with that of As5+ is probably due to the faster uptake of As3+ by endothelial cells, and inorganic arsenic exerts its toxicity at least in part via intracellular oxidative stress.