Oxidative protein damage parameters in plasma in chronic experimental diabetes in rats.

An increase in oxidative stress may contribute to the development of oxidative protein damage (OPD) in the streptozotocin-diabetic rat. To show the effect of hyperglycemia in promoting OPD, we determined protein carbonyl (PCO), nitrotyrosine (NT), total thiol (T-SH) and advanced oxidation protein product (AOPP) levels as markers of OPD, and lipid hydroperoxide (LHP) levels as a marker of lipid peroxidation in plasma of acute and chronic diabetic male Sprague-Dawley rats and their controls. The levels of the studied markers, except NT, were determined by colorimetric methods. NT levels were measured by ELISA. Plasma PCO and AOPP levels of chronic diabetic rats were increased significantly compared with those of both acute diabetic rats and the controls. Plasma NT levels of the three groups were not different. Plasma T-SH levels of acute diabetics were increased significantly compared with those of the controls while T-SH increase in the chronic diabetics was not significant. Plasma LHP levels were increased significantly in the chronic diabetic rats compared with those of the controls. The increase in plasma PCO, AOPP, LHP levels in chronic but not in acute diabetic rats may be indicating that persistence of hyperglycemia is involved in the evolution of OPD while plasma NT levels do not seem to reflect OPD in diabetes.

Relation of oxidative protein damage and nitrotyrosine levels in the aging rat brain.

An increase in oxidative stress may contribute to the development of oxidative protein damage in the aging rat brain. In the present study, we investigated the relation between nitrotyrosine levels and other oxidative protein damage parameters such as protein carbonyl and protein thiol, as well as oxidative stress parameters such as total thiol, nonprotein thiol, and lipid hydroperoxides in the brain tissue of young, adult, and old Wistar rats. Brain nitrotyrosine levels of old rats were significantly decreased compared with those of young rats. Young and adult rats were not significantly different as far as these parameters were concerned, however, brain protein carbonyl and lipid hydroperoxide levels of old rats were significantly increased compared with those of young and adult rats. On the other hand, brain tissue total thiol, nonprotein thiol, and protein thiol levels of old rats were significantly decreased compared with those of young and adult rats. The strong correlation we found between protein carbonyl and lipid hydroperoxide levels indicates a striking relation between protein oxidation and lipid peroxidation in the aging brain tissue. The results of this study suggest that protein carbonyl formation is both a sensitive and a specific marker of brain aging. However, decreased nitrotyrosine levels in old rats, in contradiction to the expected, may be due to mechanisms other than oxidative protein damage in the aging rat brain.

Changes in bone turnover and deoxypyridinoline levels in epileptic patients.

In this study, we evaluated bone turnover in 52 epileptic patients receiving chronic anticonvulsant therapy and in 39 healthy volunteers whose ages matched those of the patients. We determined serum osteocalcin and total and bone alkaline phosphatase levels as markers of bone formation, and urinary deoxypyridinoline and urinary calcium levels as markers of bone resorption. Statistical comparison of the levels of these markers between sexes in epileptic patients and their control groups revealed that total alkaline phosphatase levels were significantly increased in patients from both sexes compared with those of their controls. Urinary deoxypyridinoline levels of male epileptic patients were significantly increased compared with those of their controls. On the other hand, 25-hydroxyvitamin D levels of the male patients were significantly reduced compared with those of their controls. Serum osteocalcin, bone alkaline phosphatase, and urinary calcium levels of epileptic patients were not statistically different from those of the controls. We found that urinary deoxypyridinoline levels of male epileptic patients were increased, however, we observed no difference in serum osteocalcin and bone alkaline phosphatase levels. The lack of difference may be attributed to the fact that only the resorption phase of bone turnover is affected during chronic anticonvulsant therapy.

Effect of alpha-lipoic acid supplementation on oxidative protein damage in the streptozotocin-diabetic rat.

An increase in oxidative stress may contribute to the development of oxidative protein damage in streptozotocin diabetic rats. In the present study, the influence of alpha-lipoic acid supplementation on plasma protein carbonyl, plasma thiol, and plasma lipid hydroperoxide levels was examined in order to characterize the relationship between the oxidative stress and the oxidative protein damage. Rats were randomly divided into three groups of equal body weight. Chronic hyperglycemia was induced by intravenous streptozotocin injection in both the group of male Wistar rats to be supplemented with alpha-lipoic acid and the group that was not to receive alpha-lipoic acid. Nondiabetic rats formed the control group and received a saline injection. In streptozotocin diabetic rats with and without alpha-lipoic acid supplementation, plasma carbonyl levels were significantly increased, while plasma thiol levels were significantly decreased compared with those of the control group. Plasma lipid hydroperoxide levels were significantly increased in diabetic rats without alpha-lipoic acid supplementation compared with those of the controls, but the lipid hydroperoxide levels in the alpha-lipoic acid supplemented group were no different from those of the controls. In streptozotocin-diabetic rats, oxidative stress was significantly decreased in the alpha-lipoic acid-supplemented group. The results of this study suggest that alpha-lipoic acid, by decreasing oxidative stress, may be effective in preventing oxidative protein damage, which may contribute to the development of diabetic complications.

Oxidative protein damage in plasma of type 2 diabetic patients.

In this study, we evaluated protein oxidation in 84 patients with Type 2 diabetes with no complications and in 61 healthy volunteers who formed the control group, whose ages matched those of the patients. We determined plasma carbonyl and plasma thiol levels as markers of oxidative protein damage and erythrocyte glutathione, plasma ceruloplasmin and transferrin as markers of free radical scavengers. The concentrations (mean +/- SD) of both of plasma carbonyl (1.24 +/- 0.46 vs. 0.72 +/- 0.17 nmole/mg protein; p < 0.0001) and lipid hydroperoxides (1.8 +/- 0.63 vs. 1.3 +/- 0.21 micromole/l; p < 0.0001) were increased, and the concentration of plasma transferrin (3.85 +/- 0.65 vs. 4.59 +/- 0.79 g/l; p < 0.05) was decreased, respectively, in Type 2 diabetic patients compared with those of the controls. There were no significant differences in the concentrations of plasma thiol (0.0064 +/- 0.001 vs. 0.0068 +/- 0.001 micromole/mg protein), erythrocyte glutathione (2.54 +/- 0.57 vs. 2.65 +/- 0.56 mg/g Hb), plasma ceruloplasmin (548 +/- 107.30 vs. 609 +/- 93.34 mg/l) between the patients and the controls. These changes observed in diabetic patients contribute to the imbalance in the redox status of the plasma. We attribute this imbalance to oxidative protein damage in Type 2 diabetic patients clinically free of complications.

Effects of hormone replacement therapy on lipid peroxides and oxidation system in postmenopausal women.

A short-term evaluation of 6 months of estrogen therapy on oxidant status in 38 postmenopausal women was conducted. The levels of serum lipid peroxidation products, glutathione (GSH) status, and glutathione-related enzymes were evaluated before and after 6 months of hormone replacement therapy. After 6 months of estrogen treatment there was a significantly increased concentration of thiobarbituric acid-reactive substances (TBARS), which are an end product of lipid peroxidation. This was accompanied by a significant increase in the activity of glutathione peroxidase (GSH-Px). However, the activities of glutathione reductase (GSSG-R) and superoxide dismutase (SOD) were significantly decreased and total protein thiols were reduced. Data suggest that hormone replacement therapy in postmenopausal women is associated with oxidant mechanisms.

Changes in bone turnover on deoxypyridinoline levels in diabetic patients.

In this study, we evaluated bone turnover in 35 patients with non- insulin-dependent diabetes mellitus with no complications and in 35 healthy volunteers who formed the control group and whose ages matched those of the patients. We determined serum osteocalcin (OC) and total alkaline phosphatase levels as markers of bone formation, and urinary deoxypyridinoline (DPD) and urinary calcium levels as markers of bone resorption. Statistical comparison of the levels of these markers between sexes in diabetic and control groups revealed that the OC levels were significantly decreased in males and females in the diabetic group compared with those of the control group. No significant difference was observed for other markers. It was found that the OC levels of diabetic subjects were not affected by the type of therapy. No statistically significant difference was found in serum calcium, phosphorus and magnesium levels between the diabetic and the control groups. In our study, the finding that, in spite of the decrease in the OC levels in the diabetic group no difference was observed in DPD levels, was attributed to the fact that only the formation phase was affected in diabetes, while the resorption phase remained unaltered.