Differential effects of various trivalent and pentavalent organic and inorganic arsenic species on glucose metabolism in isolated kidney cells

Abstract

We have compared the acute toxicities of the trivalent arsenic species arsenite, oxophenylarsine (PhAsO), 2-chlorovinyloxoarsine (ClvinAsO), methyloxoarsine (MeAsO), and of the pentavalent arsenic species arsenate, methyl- and phenyl-arsonic acid in rat kidney tubules (RKT) and Madin-Darby canine kidney (MDCK) cells. In RKT, PhAsO (1 μmol I−1, 60 min) almost completely (>90%) blocked gluconeogenesis without affecting cell viability as assessed by dye exclusion. In MDCK cells, PhAsO (2 μmol I−1) markedly inhibited glucose uptake (60% of controls) within 30 min, while cell viability, as assessed by formazan formation, was not affected within 180 min. MeAsO and CIvinAsO were similarly effective to PhAsO in both RKT and MDCK cells. Estimated IC50 values for the inhibition of gluconeogenesis were 0.55 (PhAsO), 0.69 (CIvinAsO) and 0.99 μmol I−1 (MeAsO) and for the inhibition of glucose uptake 1.23 (PhAsO). 2.62 (CIvinAsO) and 6.99 μmol I−1 (MeAsO). At longer storage times, aqueous solutions of MeAsO and of CIvinAsO, but not of PhAsO, gradually lost toxic activity in RKT and MDCK cells, especially at alkaline pH. Concomitantly, a gradual decrease in content as assessed by HPLC was detected. Roughly 10-fold higher concentrations of arsenite than of PhAsO were required for comparable effects on gluconeogenesis in RKT, whereas in MDCK cells about 100-fold higher concentrations were needed for similar inhibition of glucose uptake. Pentavalent arsenate and phenylarsonate were two orders of magnitude less effective than PhAsO in RKT, while methylarsonate had virtually no influence on gluconeogenic activity. In MDCK cells the pentavalent arsenic species showed effects only in the millimolar range. It is concluded (1) that different mechanisms are involved in the acute toxicity of oxoarsines and inorganic arsenic and (2) that PhAsO offers advantages as a model substance for mono-substituted trivalent arsenicals, because it is more stable and more readily detectable.