Carbon monoxide alleviates cadmium-induced oxidative damage by modulating glutathione metabolism in the roots of Medicago sativa

Using pharmacological and biochemical approaches, the role of cadmium (Cd)-induced carbon monoxide (CO) release and the relationship between CO and oxidative stress conferred by Cd exposure in the root tissues of alfalfa (Medicago sativa) plants were investigated. Cd treatments showed a dose-dependent enhancement in lipid peroxidation. Both 100 and 200 mu M CdCl2 treatments caused the increase of CO release, which is consistent with the changes in the activity of the CO synthetic enzyme heme oxygenase (HO) and its HO-1 transcript. A 100 mu M CdCl2 exposure enhanced the formation of nonprotein thiols (NPT), and reduced glutathione (GSH) to oxidized glutathione (GSSG), which was potentiated by the pretreatment of CO scavenger hemoglobin (Hb). Plants pretreated for 6 h with 50% CO-saturated aqueous solution, which induced the rapid endogenous CO release followed by a gradual decrease when subsequently exposed to 100 mu M CdCl2 for 72 h, effectively decreased oxidative damage. Meanwhile, CO pretreatment modulated several enzymes responsible for GSH metabolism, thus resulting in the partial restoration of GSH : GSSG ratio, which was significantly blocked by Hb. These results are suggestive of a role for CO release as a signal element for the alleviation of Cd-induced oxidative damage by modulating glutathione metabolism.