Effects of preconditioning with ebselen on glutathione metabolism and stress protein expression.

Selenium induces several proteins, including glutathione and stress proteins. These proteins have been shown to be cardioprotective against oxidative injury. To determine whether ebselen, a seleno-organic compound, can also induce these proteins and exert cardioprotective action, we examined the effects of preconditioning with ebselen on glutathione metabolism and stress protein expression and on myocyte injury induced by oxidative stress. Treatment of cultured cardiac myocytes with ebselen (0.3-30 microM) for 24 hr increased the reduced glutathione content. Glutathione reductase activity, but not glutathione peroxidase activity, was significantly elevated in a dose-dependent manner. Pretreatment with ebselen increased the expression of such stress proteins as heat shock protein 70 and heme oxygenase-1 (heat shock protein 32) in cardiac myocytes, as assessed by Western blotting. Expression of heat shock protein 70 was increased only at a higher dose of ebselen (30 microM), whereas expression of heme oxygenase-1 was markedly increased at a lower dose of ebselen (3 microM). Under these conditions, the myocyte injury induced by hydrogen peroxide or simulated ischemia/reperfusion, assessed by the release of lactate dehydrogenase into the culture medium, was reduced by ebselen pretreatment in a dose-dependent manner. Results indicated that cardiac myocytes pharmacologically preconditioned with ebselen for 24 hr exhibited resistance to oxidative injury, possibly via the up-regulation of glutathione metabolism and the expression of stress proteins.