Redox homeostasis of albumin in relation to alpha-lipoic acid and dihydrolipoic acid.

Albumin represents the predominant circulating antioxidant agent in plasma exposed to continuous oxidative stress and a change in serum albumin structure accounts for its antioxidant properties. Alterations in the redox status of albumin may result in impairments of its biological properties. Alpha-lipoic acid (LA), a naturally occurring thiol compound found in virtually all species, is a potent antioxidant with high efficacy which is also involved in the chelation of metal ions, regeneration of antioxidants, and repair of oxidatively damaged proteins. In human body LA is rapidly reduced to dihydrolipoic acid (DHLA) after intake into the cell. Both, LA and DHLA are amphipathic molecules which act as antioxidants both in hydrophilic and lipophilic environments. The present study aimed to investigate the antioxidant/pro-oxidant effects of LA and DHLA due to their concentrations in metal-catalyzed protein oxidation (MCO) of human serum albumin (HSA). Progressive oxidative modification of albumin was found in MCO system by an increased content of protein hydroperoxides (POOH), protein carbonyl groups (PCO) which is the former's major breakdown product, and other protein oxidation markers such as advanced oxidized protein products (AOPP) and protein thiol groups (P-SH). The possible antioxidant protective effects of LA and DHLA were observed with 25 microM and 50 microM; DHLA being more influential. Protein oxidation parameters were found to be lower and P-SH levels seemed higher. However, prooxidant effects of both LA and DHLA came on the scene with increased concentrations of 75 microM and 100 microM where the latter seemed the most hazardous with contradicted results. It is clear that the loss of biological activity of human serum albumin by MCO system appears of medical relevance and if LA exerts similar effects seen in the present study, it is possible that cellular prooxidant activity can also result consuming this unique antioxidant in certain doses.

Variations in systemic biomarkers of oxidative/nitrosative stress and DNA damage before and during the consequent two cycles of chemotherapy in breast cancer patients.

BACKGROUND: Previous studies have suggested the importance of redox regulation in carcinogenesis. The aim of this study was to evaluate the prognostic role of altered redox homeostasis and oxidative DNA damage in patients with breast carcinoma before and during two cycles of chemotherapy. METHODS: This study included 30 patients whose serum samples were obtained on admission before treatment, and after the first and second cycles of chemotherapy, and 20 controls. We investigated serum total antioxidant status (TAS), thiobarbituric acid reacting substances (TBARS), total nitrite/nitrate (NOx), nitrotyrosine (NT), and 8-hydroxydeoxy-guanosine (8-OHdG), as well as antioxidant enzyme activities, such as glutathione peroxidase (GPx), glutathione reductase (GRx). RESULTS: TBARS, NOx, NT and 8-OHdG concentrations were significantly increased, while antioxidant enzyme activities and TAS were significantly decreased in patients when compared to controls. A concurrent increase in TBARS, NOx, NT, and 8-OHdG and a decrease in antioxidant enzyme activities and TAS were also seen after chemotherapy. No difference was observed in the second cycle of chemotherapy when compared with the first course. CONCLUSIONS: In conclusion, decreased activities of these antioxidant enzymes and low TAS concentrations observed in our study might be due to the depletion of the antioxidant defense system to combat the free radical storm produced by chemotherapy. We suggest that the increased 8-OHdG and other oxidative/nitrosative stress products that we have measured in breast cancer patients may be prognostic risk factors for the magnitude of oxidation in serum.

Antiapoptotic effect of angiotensin-II type-1 receptor blockade in renal tubular cells of hyperoxaluric rats.

In this study, we investigated the protective effect of losartan as an AT1 receptor antagonist by evaluating the expression of apoptosis-regulatory genes that contribute to the progressive damage in the renal tubules of hyperoxaluric rats. Rats were divided into 4 groups of 10 each; control (C), ethylene glycol (EG), ethylene glycol + losartan (EG + L) and Losartan (L). For 4 weeks 0.8% EG, as a precursor for oxalate, was administered to EG and EG + L and losartan (300 mg/l) was administered to groups EG + L and L. Urine and blood samples were collected for biochemical determination. Bcl-2, bax, caspase-3 and TGF-beta 1 antibodies were used for immunohistochemistry. Apoptosis was determined by TUNEL method. A marked increase in urinary oxalate levels of the rats in EG and EG + L groups was found. In the EG group a diffuse amount of oxalate crystals into the tubular lumina and interstitium in the cortex was observed. In the EG group GBM thickening, interstitial fibrosis and tubular atrophy with infiltration of mononuclear cell findings reduced in the EG + L group were presented as well. In the EG group, immunoreactivity of TGF-beta 1 was increased in glomeruli and tubuli. In the EG + L group, immunoreactivity of TGF-beta 1 was decreased compared to the EG group. Bax expression increased in the renal tubules of EG group and reduced in the EG + L group comparing to the control. In the EG + L group, the immunoreactivity of bcl-2 was increased in glomeruli. In EG + L treated group, number of caspase-3 immunopositive cells were decreased compared to all groups (P < 0.01). Apoptotic cells were increased in the EG-treated group compared to the other groups. Decreased apoptotic cell number was observed in the EG + L compared to the EG group (P < 0.01). Our findings suggest that losartan may provide a beneficial effect against tubulointerstitial damage and decrease renal tubular apoptosis caused by hyperoxaluria.

Alterations in lipid peroxidation and antioxidant status in different types of intracranial tumors within their relative peritumoral tissues.

OBJECTIVES: Elevated levels of lipid peroxidation and changes in the concentration of enzymatic and non-enzymatic antioxidant systems have been reported in various cancers, but there are very few reports available of lipid peroxidation due to oxidative stress in patients with intracranial neoplasms. The purpose of this study was to assess alterations in lipid peroxidation and antioxidant status in different types of tumors and to compare the results with their relative peritumoral tissues and compare the oxidative status in different grades of tumors. PATIENTS AND METHODS: We investigated the extent of oxidative stress and the levels of antioxidants in 16 astrocytomas and 38 other different types intracranial tumors comparing the results with their corresponding peritumoral tissues and comparing the levels in between low-grade and high-grade tumors. The extent of lipid peroxidation as evidenced by the formation of thiobarbituric acid-reacting substances (TBARS), as well as the status of the antioxidant systems such as superoxide dismutase (SOD), glutathione reductase (GR) and reduced glutathione (GSH) in tumor tissues and adjacent peritumoral tissues was estimated. The tumoral tissues were also compared as to their degrees of malignancy. RESULTS: According to our results lipid peroxidation in brain tumor tissues was enhanced compared to the corresponding adjacent peritumoral tissues. This was accompanied by a significant tumoral decrease in both enzymatic and non-enzymatic antioxidants. The low levels of antioxidants in tumor tissues, might be related to an increased use of antioxidant systems to scavenge lipid peroxides. Also a striking elevation in TBARS levels, and decrease in SOD activity and GSH levels were seen in high-grade tumors when compared with low grades. CONCLUSION: These findings emphasize a consistent difference in the level of antioxidants between the tumoral sample and its corresponding peritumoral tissue, independently of the tumoral type, and the most pivotal action would seem to minimise exposure to endogenous and exogenous sources of oxidative stress.

Effect of vitamin E therapy on oxidative stress and erythrocyte osmotic fragility in patients on peritoneal dialysis and hemodialysis.

BACKGROUND: Several medications have been tested with the aim of decreasing oxidative stress and erythrocyte osmotic fragility in patients on dialysis. The aim of the present study was to assess the influence of vitamin E therapy on oxidative stress and erythrocyte osmotic fragility in patients on hemodialysis (HD) and peritoneal dialysis (PD). METHODS: This was a placebo-controlled study. The study was performed on 34 HD patients, 13 PD patients and 22 healthy volunteers with a mean age of 45.57 +/- 8.54 years. HD patients were divided into 2 groups: treatment (n=19) and control (n=15). Vitamin E was administered, 300 mg/day, to the HD treatment group and PD patients for 20 weeks. Lipid peroxidation, antioxidant condition and erythrocyte osmotic fragility (EOF) were examined before and after treatment. RESULTS: Before the treatment, the levels of EOF (p<0.001) and malondialdehyde (MDA) (p<0.001) were significantly lower, and erythrocyte superoxide dismutase (SOD) (p=0.001) and vitamin E levels (p<0.001) were significantly higher in the healthy group than PD and HD groups. Serum vitamin E increased from 0.93 +/- 0.16 to 1.09 +/- 0.14 mg/dL (p=0.001), EOF decreased from 0.49% +/- 0.03% to 0.42% +/- 0.04% NaCl (p<0.001), and plasma MDA values decreased from 2.77 +/- 0.87 to 2.20 +/- 0.767 nmol/mL (p=0.018) in the HD treatment group after vitamin E treatment. Levels of EOF decreased from 0.51% +/- 0.09% to 0.43% +/- 0.03% NaCl in the PD treatment group after vitamin E treatment (p=0.021). CONCLUSION: Vitamin E therapy is effective in decreasing the levels of EOF in patients on HD and PD, and it is also effective in decreasing lipid peroxidation in patients on HD.

Changes of nitric oxide synthase-containing nerve fibers and parameters for oxidative stress after unilateral cavernous nerve resection or manuplation in rat penis.

After pelvic surgeries such as radical prostatectomy, two major complications--urinary incontinence and erectile dysfunction (ED) may occur. Etiologies for ED are multiple pathologic mediators/systems. Oxidative stress, which is known to be induced after surgical trauma, could be a cause of ED. The purposes of in this study are to investigate the effect of unilateral manipulation/ dissection and resection of the cavernous nerve (neurotomy) to NOS (nitric oxide synthase)-containing nerve fibers and pressure after electro stimulation in rat corpus cavernosum, and to determine whether these procedures would produce oxidative stress within rat cavernous tissue 3 weeks and 6 months after the operation. Male rats were divided into 5 groups. Rats in groups 1 and 2 underwent unilateral cavernous nerve manipulation and sacrificed 3 weeks and 6 months after the operation, respectively. Rats in groups 3 and 4 underwent unilateral neurotomy of a 5-mm. segment of the cavernous nerve, and they were sacrificed 3 weeks and 6 months after nerve ablation, respectively. Group 5 rats were control animals for biochemical analysis. Intracavernous pressure following electro stimulation reduced is significantly 3 weeks after unilateral resection, as compared to that of the manipulated nerve (P < 0.05), and it recovered 6 months after neurotomy. The recovery was also confirmed by NADPH (nicotinamide adenine dinucleotide phosphate) diaphorase staining in neurotomy groups. Lipid peroxidation, which is an indicater of oxidative stress, was determined by measuring thiobarbituric acid reacting substance (TBARS) levels and superoxide dismutase (SOD) activity. These markers indicated that unilateral cavernous nerve manipulation or resection produced oxidative stress within rat corpus cavernosum. Oxidative stress was more prominent 3 weeks after unilateral neurotomy (P < 0.05). Also, compared to the control animal group, oxidative stress was observed three weeks after manipulation of unilateral cavernous nerve (P < 0.05). Resection of the cavernous nerve caused more prominent oxidative stress than in the manipulation group. This study suggested, that unilateral cavernous neurotomy caused a decrease of intra cavernous pressure and NOS fibers in rat corpus cavernosum, and they recovered 6 months after neurotomy. Our data also provided evidence that neurotomy and manipulation of the cavernous nerve caused oxidative stress in rat corpus cavernosum and that oxidative stress was more prominent in the nerve resection group.

Inhibition of inducible nitric oxide synthase in murine visceral larva migrans: effects on lung and liver damage.

The roles of nitric oxide production and oxidative process were studied in mice infected with Toxocara canis and treated with aminoguanidine which is a specific inhibitor of inducible nitric oxide synthase (iNOS). Relations of nitric oxide synthase inhibition and tissue pathology were assessed by biochemical, histological and immunohistochemical methods. In experiments, Balb/c albino mice were inoculated with T. canis eggs either with or without aminoguanidine treatment or alone, at 24th, 48th hours and on 7th days. LPx and SOD values in liver tissue and plasma were measured. Liver and lung tissues were evaluated for the pathological lesions. The expression of eNOS and iNOS in both tissues were studied with immunohistochemistry in the same intervals. We observed significant differences between T. canis infected and aminoguanidine treated animals. Larval toxocarosis led to oxidative stress elevation in plasma. Microscopic examination of the liver histological sections revealed pathological lesions in the hepatic parenchyma in infected mice. In the mice received T. canis eggs plus aminoguanidine, the sinusoidal areas were enlarged. Histological lesions were more severe at 48 hours after infection. Numbers of eNOS and iNOS expressing epithelial cells were increased in the T. canis infected mice. The activities of eNOS and iNOS were also observed in the body of the larvae which have migrated to lung and liver. As a result, we have demonstrated that in vivo production of eNO and iNO during T. canis infection cause direct host damages and it is strongly related to the oxidative stress. We propose that larval NO can also be effective in larval migration, but it needs further investigation on distribution of NO in larvae.

Effects of melatonin on plasma S-nitrosoglutathione and glutathione in streptozotocin-treated rats.

The aim of this study was to determine the effects of streptozotocin-induced diabetes on plasma reduced glutathione (GSH) and S-nitrosoglutathione (GSNO) levels. Further, the study investigated whether an antioxidant, pineal hormone melatonin, could protect against STZ-induced effects. STZ significantly decreased plasma GSH but increased the levels of plasma GSNO. Daily supplementation with melatonin restored plasma thiol to control values. Data suggest that STZ-induced hyperglycemia and compounds that act as scavengers of free radicals and peroxynitrite like melatonin may exert protection against STZ-induced toxicity.

In vivo inhibition of inducible nitric oxide and evaluation of the brain tissue damage induced by Toxocara canis larvae in experimentally infected mice.

Nitric oxide (NO) is known to be produced by macrophages, endothelial cells and neurons and synthesized by an enzyme called nitric oxide synthase (NOS). Various effector mechanisms and infections can affect the NO production. Excessive amount of NO will lead to biochemical reactions, which cause toxic effects. In this study the role of NO has been evaluated in larval toxocarosis, which is a systemic parasite infection caused by T. canis larvae. Infection was established in the Balb/c mice with or without inducible NOS (iNOS) inhibition and the effects of infection and NOS inhibition were observed according to the results of SOD and LPx measurements in brain tissue and NADPH-diaphorase (NADP-d) histochemistry. Results of NADPH-d histochemistry indicate that iNOS inhibition has protective effect on the brains of infected mice and that larval T. canis infection could be related to oxidative stress, and NO production and iNOS inhibition can protect the tissue from damage in this infection.

Effect of regular training on plasma thiols, malondialdehyde and carnitine concentrations in young soccer players.

Physical activity is known to induce oxidative stress in individuals subjected to intense exercise. Contrarily, there are enzymatic and nonenzymatic defence systems against oxygen radicals in aerobic organisms. Sulphydryl groups such as thiol and glutathione (GSH) can be given as an example to non-enzymatic low molecular weight antioxidants. Carnitine may be related to the performance enhancement in high intensity intermittent exercises and might probably improve the aerobic capacity by stimulating lipid oxidation in muscle cells during long term exercise. But, the effects caused by this supplement during physical activity have not been fully described in the literature. The aim of the study was to compare plasma thiols (PSH), malondialdehyde (MDA) and carnitine levels and maximal oxygen uptake (VO2(max)) of the soccers under regular training with the values of the healthy controls. Our results demonstrates that soccers seem to be under less oxidative stress, as their MDA levels were significantly lower (P < 0.001) when compared with the control group while their PSH levels were significantly elevated (P < 0.001), during resting condition. In addition, the plasma carnitine concentrations of the soccer group yields lower values while the VO2(max) yields a higher value when compared with the control group. The differences between the soccer and the control groups are significant (for both, P < 0.001). The present research reveals the fact that regular soccer training shows beneficial effect on decreasing of lipid peroxidation levels. Furthermore; the sportsmen who are under intense training programs have low plasma carnitine values which do not cause negative effect on their sportive performance.

Lipid peroxidation, erythrocyte superoxide-dismutase activity and trace metals in young male footballers.

Physical training is known to induce oxidative stress in individuals subjected to intense exercise. In this study, we investigated plasma malondialdehyde (MDA) levels and erythrocyte superoxide dismutase (SOD) activity of 25 young male footballers and a control group of similar age. Red blood cell (RBC) count, haemoglobin (Hb) and haematocrit (Hct) values, and copper (Cu) and zinc (Zn) levels were also examined. The maximal oxygen uptake (VO2max) of all subjects was determined in order to establish their functional capacity. The main finding of the present study was that plasma MDA levels, one of the most commonly used markers of lipid peroxidation, of this group of footballers aged under 21 decreased slightly when compared with those of the control group (p < 0.001). In contrast, erythrocyte SOD activity was higher in the footballer group than in the controls (p < 0.001). Footballers who are under regular training showed an improved antioxidant activity in comparison to sedentary controls. Plasma copper concentration, RBC count and Hb concentration of the footballer group were all significantly lower than those of the control group, (p < 0.001, p < 0.01, p < 0.01, respectively). Investigating the footballers' data with Spearman's correlation analyses, the correlation coefficients (r) between Zn/Cu ratio and SOD was positive (r=0.44; p < 0.05); and between VO2max and SOD (r=0.42; p < 0.05) were both positive. On the basis of statistical analysis, we suggest that regular exercise may be beneficial in cases of oxidative damage by reducing the amount of lipid peroxidation and increasing the activity of the antioxidant enzyme SOD.

The effect of vitamin E treatment on oxidative stress generated in trained rats.

The aim of this study was to investigate the effect of vitamin E treatment on increased oxidative stress in rats exposed to a swimming exercise protocol. In order to examine the effects of physical swimming training on the antioxidant defences of tissues and on their susceptibility to damage induced by exercise, the levels of glutathione (GSH) and thiobarbituric acid reacting substances (TBARS) levels, on indicator of lipid peroxidation in various tissues, have been determined. In this study, four groups of female rats were used while the rats were trained to swim for 30 minutes a day and five days a week which lasted eight weeks and vitamin E (vit. E) supplementation (30 mg/kg/day) has been carried out for five days a week. TBARS levels are significantly found lower in both trained and sedentary vit. E supplemented groups, since vit. E is the most important antioxidant in an earlier line of defence in lipid peroxidation. Also, in vit. E supplemented trained rats, the glutathione response is observed to be significantly higher, supporting with the TBARS levels and in accordance with the literature. But in the sedentary group without vit. E supplementation, the GSH levels of the liver and the heart tissues were significantly lower than both vit. E supplemented sedentary and trained groups. These results evaluate that vit. E confers protection to GSH levels in these tissues where the GSH levels were found significantly lower in the groups not supplemented with vit. E.

Melatonin administration affects plasma total sialic acid and lipid peroxidation levels in streptozotocin induced diabetic rats.

Administration of streptozotocin (STZ) was used to induce diabetes, as the mechanism involved is believed associated with generation of free radicals. Supplementation with antioxidant molecules such as melatonin may serve as a protection against diabetes. The aim of this study was to determine whether the STZ-induced effects on plasma thiobarbituric acid-reactive substances (TBARS, a marker of lipid peroxidation) and total sialic acid levels could be blocked by melatonin. STZ significantly increased the plasma levels of sialic acid and TBARS. Treatment with melatonin markedly reduced the STZ-induced effects on plasma sialic acid and TBARS and was associated with restoration of hyperglycemia to control blood glucose levels. These data suggest that melatonin protects against oxidative damage, and daily supplementation with melatonin may be beneficial for diabetics.

Vitamin E supplementation in streptozotocin-treated rats alters cerebellar and plasma nitric oxide metabolism.

Nitric oxide (NO) free radicals appear to contribute to the pathogenesis of a number of disorders including diabetes mellitus. The aim of this study was to determine the effects of streptozotocin (STZ)-induced diabetes on nitric oxide (NO) metabolites in plasma and cerebellar nitric oxide synthase (NOS) activity. Further, it was of interest to determine whether an antioxidant, vitamin E, could reverse the STZ-induced effects. STZ significantly decreased cerebellar NOS but increased the level of plasma total nitrite + nitrate and the level of plasma nitrate. Supplementation with vitamin E effectively reduced the STZ-induced effects. Data demonstrate that vitamin E may serve as a protective antioxidant in STZ-induced diabetes.