Is thioredoxin reductase involved in the defense against DNA fragmentation in varicocele?

We aimed to investigate the role of thioredoxin reductase (TR) and inducible heat shock protein 70 (iHsp70) and their relationship with sperm quality in varicocele (VAR) patients. Semen samples were obtained from 16 subfertile men diagnosed as VAR and 10 fertile men who applied to the Andrology Laboratory of Istanbul Medical Faculty of Istanbul University. The sperm TR and iHsp 70 expression levels were determined using Western blot analysis. The TR activity of the sperm was assayed spectrophometrically. The sperm quality was evaluated both by conventional sperm analysis and by a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) technique that assayed DNA-fragmented spermatozoa in semen samples. The percentage of TUNEL-positive spermatozoa in the VAR group (16.3%± 5.6%) was higher than that in the fertile group (5.5%± 1.9%). Significant inverse correlations were detected between the percentage of TUNEL-positive cells and both the concentration (r=-0.609; P=0.001) and motility (r=-0.550; P=0.004) of spermatozoa. Both the TR expression and activity were increased significantly in the VAR group (U=22.0; P=0.001 and U=33.5; P=0.012, respectively) as analyzed using the Mann-Whitney U Wilcoxon rank sum W test. Furthermore, significant positive correlations were found between TR expression and activity (r=0.406; P=0.040) and between TR expression and the percentage of TUNEL-positive cells (r=0.665; P=0.001). Sperm iHsp70 expression did not differ between the VAR and fertile groups. In conclusion, increased sperm TR expression might be a defense mechanism against apoptosis in the spermatozoa of men with VAR.

Mystery of idiopathic male infertility: is oxidative stress an actual risk?

OBJECTIVE: To study the role of oxidative stress in sperm dysfunction in Turkish idiopathic infertile men. DESIGN: Prospective study. SETTING: Medical laboratory. PATIENT(S): Semen samples from 28 idiopathic infertile men and 14 fertile men. INTERVENTION(S): Sperm DNA fragmentation and reactive oxygen species (ROS) formation were assayed with the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) test and 2',7'-dichlorodihydrofluorescein, respectively. Seminal plasma protein carbonyl groups (PC), nitrotyrosine (NT), malondialdehyde (MDA), and total thiol (SH) levels and ferric reducing antioxidant power (FRAP) were determined. MAIN OUTCOME MEASURE(S): Sperm DNA fragmentation in relation to ROS formation and seminal plasma oxidative parameters. RESULT(S): The number of TUNEL-positive spermatozoa from idiopathic infertile men was higher than from fertile men, and ROS formation was increased as well in infertile males. A positive correlation was detected between TUNEL-positive cells and ROS content. Seminal plasma MDA, PC, and NT levels were elevated in idiopathic infertile males. No difference was observed in the total SH content and FRAP. Seminal plasma MDA levels correlated positively with both NT and PC levels. Positive correlations were detected between DNA fragmentation and MDA, NT, and PC of seminal plasma, and between sperm ROS content and MDA levels. CONCLUSION(S): The results of this study support the presence of oxidative stress in sperm dysfunction in Turkish idiopathic infertile men.

Effect of artichoke leaf extract on hepatic and cardiac oxidative stress in rats fed on high cholesterol diet.

Hypercholesterolemia and lipid peroxidation play complementary roles in atherosclerosis. Artichoke (Cynara scolymus L., Asteraceae) leaf extract (ALE), rich in antioxidants, has cholesterol-reducing effect. We investigated the effect of ALE on serum and hepatic lipid levels and pro-oxidant-antioxidant balance in the liver and heart of hypercholesterolemic rats. Rats were fed on 4% (w/w) cholesterol and 1% cholic acid (w/w) supplemented diet for 1 month. ALE (1.5 g/kg/day) was given by gavage during the last 2 weeks. High cholesterol (HC) diet caused significant increases in serum and liver cholesterol and triglyceride levels. It increased malondialdehyde (MDA) and diene conjugate (DC) levels in both tissues. Hepatic vitamin E levels and hepatic and cardiac glutathione peroxidase (GSH-Px) activities decreased, but superoxide dismutase and glutathione transferase activities, glutathione, and vitamin C levels remained unchanged due to HC diet. Serum cholesterol and triglyceride levels and ratio of cholesterol to high-density lipoprotein (HDL)-cholesterol decreased in ALE plus HC-treated rats, but liver cholesterol and triglyceride levels remained unchanged. Significant decreases in hepatic and cardiac MDA and DC levels and increases in hepatic vitamin E and GSH-Px activities were observed in ALE-treated hypercholesterolemic rats. Our results indicate that ALE decreases serum lipids and hypercholesterolemia-induced pro-oxidant state in both tissues.

Effect of taurine treatment on pro-oxidant-antioxidant balance in livers and brains of old rats.

The effect of taurine treatment on antioxidant defense in liver and brain tissues of old rats was investigated. Endogenous malondialdehyde (MDA) and diene conjugate (DC), ascorbic acid (AA)- and NADPH-induced lipid peroxide levels as well as non-enzymatic (glutathione--GSH, vitamin E and vitamin C) and enzymatic antioxidants (superoxide dismutase, glutathione peroxidase and glutathione transferase) were determined in livers and brains of young (5 months), old (22 months), and taurine-treated old rats. Taurine (2%, w/v; in drinking water) was administered to old rats for 6 weeks. Taurine levels decreased in the liver and brain of old rats compared to young rats.MDA and DC levels increased, GSH levels decreased, but induced lipid peroxidation remained unchanged in livers of aged rats. Oxidative stress parameters did not change in brains of aged rats. Taurine treatment resulted in significant increases in taurine levels, decreases in MDA and DC levels and increases in GSH levels in livers of old rats. Taurine treatment also increased brain taurine levels. However, no significant changes were detected in lipid peroxidation and antioxidant system in brains of old rats following taurine treatment. Accordingly, in old rats the liver seems more susceptible to age-related lipid peroxidation increases and taurine level changes than the brain. Thus, taurine supplementation seems to be useful for decreasing hepatic oxidative stress in aging.

Induced oxidative stress and decreased expression of inducible heat shock protein 70 (ihsp 70) in patients with colorectal adenocarcinomas.

BACKGROUND: Colorectal cancer is one of the most common carcinomas observed in humans. Recently we have reported that increased oxidative stress is associated with human colorectal cancer. There are few and controversial studies on the clinical relevance of the expression of heat shock protein 70 (HSP 70), a member of the HSP family, in colorectal cancer. In this study, we assayed lipid peroxide levels, glutathione peroxidase (GPx) activity and the expression of inducible heat shock protein 70 (ihsp 70) in 20 paired samples of colorectal tumor and adjacent normal tissues in order to determine the relationship between oxidative stress and ihsp 70 expression. Histopathological results were also considered to establish the clinical relevance of ihsp 70 in colorectal cancer. METHODS: Malondialdehyde (MDA) levels as an indicator of lipid peroxidation and GPx activity were assayed by spectrophotometric methods. The Western blotting procedure was used for the determination of ihsp 70 expression. RESULTS: Significant increases were observed in MDA levels (111%) and GPx activities (50%) of malignant tissues as compared with normal tissues of the patients with colorectal cancer. The expression of ihsp 70 was found to be decreased in malignant tissues as compared with normal tissues of the patients. Significant negative correlations were detected between MDA levels and ihsp expression (r = -0.516; P < 0.05 ) and GPx activity and ihsp 70 expression (r = -0.471; P < 0.05) in malignant tissues of patients. When the patients were grouped according to histopathological characteristics, no difference was found in MDA levels, GPx activity and ihsp 70 expression. CONCLUSION: Our results indicate that ihsp70 expression is suppressed under induced oxidative stress conditions in malignant tissues of patients with colorectal cancer. Further research is needed to clarify the mechanisms responsible for this decrease and the definitive role of ihsp 70 in colorectal cancer.