The detection of S-glutathionation of hepatic carbonic anhydrase III in rats treated with paraquat or diquat.

Protein S-glutathionation has been demonstrated to be one of the cellular responses under oxidative stress and may be involved in many cellular metabolisms. In this study, the effect of redox cycling bipyridylium compounds, paraquat and diquat, on this protein modification was investigated. Male Sprague-Dawley rats were administered i.p. either paraquat at 20 or 40 mg/kg body wt. or diquat at 85 or 170 mg/kg body wt., respectively. The liver was examined at different time points for taking the measurement of the S-glutathionation of carbonic anhydrase III (CA III), thiobarbituric acid-reactive substances (TBARS), vitamin E depletion, glutathione (GSH) and glutathione disulfide (GSSG) contents. The extent of S-glutathionation of CA III was chosen as a marker and was determined by a method combining isoelectric focusing analysis with immunoblotting. Those results indicated that paraquat and diquat significantly increased the generation of TBARS and showed a time-dependent response. The significant effect on vitamin E depletion was only obtained in rats treated with a high dose of diquat for 2 h. Hepatic cellular GSSG contents did not increase but tended to decrease all of the treatments. Although oxidative damage was actually generated in liver, based on the increase of TBARS generation and vitamin E depletion, no increase of CA III S-glutathionation was observed. We propose that the reason for this observation under this circumstance is probably due to the reversible characteristic of CA III S-glutathionation, which has been demonstrated in our previous study (Chai et al., 1991) Arch. Biochem. Biophys. 384, 270-278) and named as dethiolation.