Heme oxygenase induction by menadione bisulfite adduct-generated oxidative stress in rat liver.

The in vivo effect of menadione bisulfite adduct on both hepatic oxidative stress and heme oxygenase induction was studied. A marked increase in lipid peroxidation was observed 1 h after menadione bisulfite adduct administration. To evaluate liver antioxidant enzymatic defenses, superoxide dismutase, catalase and glutathione peroxidase activities were determined. Antioxidant enzymes significantly decreased 3 h after menadione bisulfite adduct injection. Heme oxygenase activity appeared 6 h after treatment, peaking 9 h after menadione bisulfite adduct administration. Such induction was preceded by a decrease in the intrahepatic GSH pool and an increase in hydrogen peroxide steady-state concentration, both effects taking place some hours before induction of heme oxygenase. Iron ferritin levels and ferritin content began to increase 6 h after heme oxygenase induction, and these increases were significantly higher 15 h after treatment and remained high for at least 24 h after menadione bisulfite adduct injection. Administration of bilirubin entirely prevented heme oxygenase induction as well as the decrease in hepatic GSH and the increase in lipid peroxidation when administered 2 h before menadione bisulfite adduct treatment. These results indicate that the induction of heme oxygenase by menadione bisulfite adduct may be a general response to oxidant stress, by increasing bilirubin and ferritin levels and could therefore provide a major cellular defense mechanism against oxidative damage.

Effect of acetaminophen on heme metabolism in rat liver.

BACKGROUND AND AIMS: Acetaminophen (APAP) or paracetamol is a hepatotoxic drug through mechanisms involving oxidative stress. To know whether mammalian cells possess inducible pathways for antioxidant defense, we have to study the relationship between heme metabolism and oxidative stress. METHODS: fasted female Wistar rats received a single injection of APAP (3.3 mmol kg(-1) body weight) and then were killed at different times. Heme oxygenase-1 (HO), delta-aminolevulinic acid (ALA) synthase, ALA dehydratase, and porphobilinogenase activities, lipid peroxidation, GSH, catalase and glutathione peroxidase, were measured in liver homogenates. The antioxidant properties of bilirubin and S-adenosyl-L-methionine were also evaluated. RESULTS: APAP increased lipid peroxidation (115% +/- 6; S.E.M., n=12 over control values) 1 h after treatment. GSH reached a minimum at 3 h (38% +/- 5) increasing thereafter. At the same time antioxidant enzymes reached minimum values (catalase, 5. 6 +/- 0.4 pmol mg(-1) protein, glutathione peroxidase, 0.101 +/- 0.006 U mg(-1) protein). HO induction was observed 6 h after treatment reaching a maximum value of 2.56 +/- 0.12 U mg(-1) protein 15 after injection. ALA synthase (ALA-S) induction occurred after enhancement of HO, reaching a maximum at 18 h (three-fold the control). ALA dehydratase activity was first inhibited (31 +/- 3%) showing a profile similar to that of GSH, while porphobilinogenase activity was not modified along the whole period of the assay. Administration of bilirubin (5 micromol kg(-1) body weight) or S-adenosyl L-methionine (46 micromol kg(-1) body weight) 2 h before APAP treatment entirely prevented the increase in malondialdehyde (MDA) content, the decrease in GSH levels as well as HO and ALA-S induction. CONCLUSION: This study shows that oxidative stress produced by APAP leads to increase in ALA-S and HO activities, indicating that toxic doses of APAP affect both heme biosynthesis and degradation.

Heme oxygenase induction by UVA radiation. A response to oxidative stress in rat liver.

Heme oxygenase is a key enzyme for heme catabolism and catalyzes the oxidative degradation of heme to form biliverdin IX alpha, an immediate precursor of bilirubin. In order to shed light on the mechanism by which UVA radiation causes oxidative damage, the relationship between heme oxygenase induction and oxidative stress was studied. HO-1 activity, lipid peroxidation and generation of active oxygen species (H2O2) were measured in rat liver exposed to UVA radiation. Besides, soluble and enzymatic antioxidant defenses (GSH, SOD, CAT and GSH-Px) were determined, while bilirubin antioxidant capacity was also evaluated. UVA radiation markedly increased both lipid peroxidation (180% +/- 7; S.E.M., n = 9 over control value of 0.1 +/- 0.01 nmol MDA/min per mg prot.) and steady state concentration of hydrogen peroxide (4 +/- 0.03 microM; S.E.M., n = 9) 3 h after treatment. At the same time, GSH content decreased to 3.6 +/- 0.2 mumol/g liver (S.E.M., n = 9) increasing thereafter. Antioxidant enzymes reached minimum values 6 h after UVA treatment (SOD: 7.2 +/- 0.2 U/mg protein, CAT: 7.8 +/- 0.2 pmol/mg protein, GSH-Px: 0.088 +/- 0.004 U/mg protein; S.E.M., n = 9), starting to increase 12 h after irradiation. HO-1 induction was observed 6 h after UVA irradiation, reaching a maximum value of 2.5 +/- 0.03 U/mg protein (S.E.M., n = 9) 12 h after treatment, and then declined until it reached control levels 24 h after exposure. Administration of bilirubin 2 h before UVA irradiation, entirely prevented HO-1 induction, the increase in MDA content and the decrease in GSH levels. This study shows that UVA irradiation leads to oxidative stress as evidenced by increased MDA content and H2O2 steady state levels, and depletion of GSH, SOD, CAT and GSH-Px. All these changes produced HO-1 induction. It is concluded that the induction of this enzyme could be a response to oxidative stress, since bilirubin can act as a physiological antioxidant.

Relationship between oxidative stress and heme oxygenase induction by copper sulfate.

The effect of copper sulfate (CuSO4) on both hepatic oxidative stress and heme oxygenase induction was studied. A strong increase in in vivo rat liver chemiluminescence was observed 1 h after Cu(II) administration. To evaluate liver antioxidant enzymatic defenses, superoxide dismutase, catalase, and glutathione peroxidase activities were determined. Catalase and glutathione peroxidase were found to be significantly decreased 5 h after CuSO4 injection. In contrast, superoxide dismutase activity was increased. Heme oxygenase activity appeared 5 h after treatment, reaching a maximum value 18 h after CuSO4 administration. This induction was preceded by a decrease in the intrahepatic GSH pool and an increase in the generation of thiobarbituric acid reactive substances, both effects taking place a number of hours before induction of heme oxygenase. Administration of bilirubin, the end product of heme catabolism in mammals, and alpha-tocopherol, a widely employed antioxidant, completely prevented heme oxygenase induction as well as the decrease in hepatic GSH and the increase in chemiluminescence when administered 2 h before CuSO4 treatment. Under the same experimental conditions, beta-carotene showed a moderate preventive effect on both heme oxygenase induction and oxidative stress parameters. These data obtained with Cu(II) treatment are in agreement with our previous reports suggesting a correlation between heme oxygenase induction and oxidative stress.

Heme oxygenase induction by cadmium chloride: evidence for oxidative stress involvement.

Cadmium chloride (CdCl2), a well-known inducer of heme oxygenase, produced a strong increase in 'in vivo' rat liver chemiluminescence (QLV) 3 h after administration. Heme oxygenase activity increased 5 h after treatment, reaching a maximum value around 12-15 h after CdCl2 administration. Such induction was preceded by a decrease in the intrahepatic GSH pool and an increase in hydrogen peroxide steady-state concentration, both effects taking place several hours before induction of heme oxygenase. The activity of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) was found to be significantly decreased 5 h after CdCl2 injection. Administration of bilirubin, the end product of heme catabolism in mammals, and alpha-tocopherol, a widely employed antioxidant, prevented heme oxygenase induction as well as the decrease in hepatic GSH and the increase in chemiluminescence when administered 2 h before CdCl2 treatment. These results obtained with CdCl2 treatment support our recent reports correlating heme oxygenase induction with oxidative stress.