Effect of Lipoic Acid on Serum Paraoxonase-1 and Paraoxonase-3 Protein Levels and Activities in Diabetic Rats.

The aim of the present study was to investigate the effect of streptozotocin-induced diabetes mellitus and lipoic acid treatment on serum paraoxonase-1 and paraoxonase-3 protein levels and paraoxonase, arylesterase and lactonase activities.36 rats were equally and randomly divided into 4 groups as control, lipoic acid, diabetes and diabetes+lipoic acid. To induce diabetes, a single dose of streptozotocin (40 mg/kg) was injected intraperitoneally to diabetes and diabetes+lipoic acid groups. Lipoic acid (10 mg/kg/day) was injected intraperitoneally for 14 days to lipoic acid and diabetes+lipoic acid groups. Serum PON1 and PON3 protein levels were measured by western blotting. Serum paraoxonase, arylesterase and lactonase activities were determined by the measuring initial rate of substrate (paraoxon, phenylacetate and dihydrocoumarin) hydrolysis.Streptozotocin-induced diabetes mellitus caused a significant decrease whereas lipoic acid treatment caused a significant increase in serum PON1 and PON3 protein levels and paraoxonase, arylesterase and lactonase activities. The increase percent of serum PON3 protein was higher than that of serum PON1 protein and the increase percent of serum lactonase activity was higher than that of serum paraoxonase and arylesterase activities in diabetes+lipoic acid group.We can report that, like PON1 protein, PON3 protein and actually its lactonase activity may also have a role as an antioxidant in diabetes mellitus and lipoic acid treatment may be useful for the prevention of the atherosclerotic complications of diabetes by increasing serum PON1 and PON3 protein levels and serum enzyme activities.

The effect of lipoic acid in the prevention of myocardial infarction in diabetic rats.

OBJECTIVES: We aimed to investigate the effect of lipoic acid in the prevention of myocardial infarction in diabetic rats. METHODS: Rats were divided into five groups as control, ISO, LA+ISO, STZ+ISO and STZ+LA+ISO. To induce diabetes, single dose of streptozotocin was injected to STZ+ISO and STZ+LA+ISO groups. Lipoic acid (10 mg/kg/day) was injected for 14 days to LA+ISO and STZ+LA+ISO groups. To induce myocardial infarction, isoproterenol was injected to ISO, LA+ISO, STZ+ISO and STZ+LA+ISO groups on the days 13 and 14 of lipoic acid treatment. Cardiac necrosis and leucocyte infiltration were investigated histopathologically. Serum malondialdehyde levels, paraoxonase and lactonase activities were measured spectrophotometrically. RESULTS: Isoproterenol caused a significant increase in cardiac necrosis, leucocyte infiltration and serum lipid peroxidation whereas a significant decrease in serum paraoxonase and lactonase activities. In myocardial infarcted non-diabetic rats, while lipoic acid caused a significant decrease in cardiac necrosis, leucocyte infiltration and serum lipid peroxidation and a significant increase in serum paraoxonase and lactonase activities, it did not change these histopathologic or biochemical parameters in myocardial infarcted diabetic rats. CONCLUSION: Lipoic acid, at the dose of 10 mg/kg, is effective to prevent myocardial infarction in non-diabetic rats but it is insufficient in diabetic rats (Tab. 1, Fig. 2, Ref. 35).

Can radiation-induced chronic oxidative stress in kidney and liver be prevented by dimethylsulfoxide? Biochemical determination by serum and tissue markers.

PURPOSE: To investigate the protective effects of dimethylsulfoxide (DMSO) on chronic oxidative stress in the liver, kidney and serum with biochemical parameters such as malondialdehyde (MDA), advanced oxidation protein product (AOPP), catalase, glutathione (GSH), and free-thiols (F-SH). METHODS: Thirty Wistar albino female rats were randomly divided into 3 groups: group I (control, n=10), group II (irradiation-alone group, n=10) and group III (DMSO and irradiation group, n=10). Rats in groups II and III were irradiated with a single dose of 6 Gy to the entire liver and right kidney. Group III received DMSO 4.5 g/kg by intraperitoneal injection 30 min before irradiation. At the end of the 24th week, the rats were sacrificed and their trunk blood, kidney and liver tissues were collected. RESULTS: Group II rats showed increased levels of lipid peroxidation and protein oxidation, with decreased GSH, FSH and catalase levels in all specimens when compared with group I. Serum and kidney MDA and AOPP levels were significantly lower in group III when compared with group II. However, serum and kidney GSH and F-SH levels were significantly higher in group III when compared with group II. The additive effect on catalase was seen only in the serum. CONCLUSION: DMSO is a protective agent on chronic oxidative stress in the serum and kidney tissue. No oxidant or antioxidant effect of DMSO in the liver was seen.

The role of N-acetylcysteine in lower extremity ischemia/reperfusions.

AIM: To evaluate the efficacy of N-acetyl cysteine (NAC) in lower extremity ischemia/reperfusion. METHODS: A total of 23 patients who underwent surgical intervention due to acute femoral artery occlusion were assigned into 2 groups: control group (group 1, n=12); and NAC group (group 2, n=11). Patients in NAC group received NAC before reperfusion, and 8 and 16 h after reperfusion (3x300 mg), while patients in control group received only NaCl 0.9% (3x100 mL). Catalase, malondialdehyde (MDA) and thiol concentrations were determined in femoral vein samples collected at 6 different time points: before reperfusion (t1), and 30 min (t2), 2 h (t3), 6 h (t4), 12 h (t5) and 24 h (t6) after reperfusion. Alveolar-arterial oxygen gradient (A-aO2) was calculated in radial artery blood samples simultaneously collected at the same time points. RESULTS: No significant differences between the two groups with regard to age (control group 61+/-13 and NAC group 64+/-11 years), gender (control group M/F: 7/5, NAC 6/5) and the average time from onset of symptoms (control group 9.6+/-3.5 h, and NAC group 10.2+/-3.1 h) were present. Catalase enzyme activity increased with reperfusion in both groups and there were no differences between the two groups. MDA levels did not change significantly with reperfusion in NAC group, whereas they were significantly higher in control group at t2 and t3 compared to NAC group (P<0.05). Thiol concentrations decreased with reperfusion in control group, and in NAC group increases that started with reperfusion returned back to baseline levels after 24 hours. Although the A-aO2 gradient increased in both groups with the beginning of reperfusion, the most prominent increase occurred in control group (P<0.05). CONCLUSIONS: In control group, the significant increase in MDA levels and A-aO2 gradient in reperfusion phase were considered a sign of local and end organ injury. We did not observe these changes in NAC performed group thus showing the efficacy of NAC.

Effect of psychological stress on the L-arginine-nitric oxide pathway and semen quality.

It has been reported that mental stress causes abnormality of spermiogram parameters. We investigated the effect of psychological stress on the L-arginine-nitric oxide (NO) pathway. Semen samples were collected from 29 healthy fourth semester medical students just before (stress) and 3 months after (non-stress) the final examinations. Psychological stress was measured by the State Anxiety Inventory questionnaire. After standard semen analysis, arginase activity and NO concentration were measured spectrophotometrically in the seminal plasma. Measurements were made in duplicate. During the stress period, sperm concentration (41.28 +/- 3.70 vs 77.62 +/- 7.13 x 10(6)/mL), rapid progressive motility of spermatozoa (8.79 +/- 1.66 vs 20.86 +/- 1.63%) and seminal plasma arginase activity (0.12 +/- 0.01 vs 0.22 +/- 0.01 U/mL) were significantly lower than in the non-stress situation, whereas seminal plasma NO (17.28 +/- 0.56 vs 10.02 +/- 0.49 micromol/L) was higher compared to the non-stress period (P < 0.001 for all). During stress there was a negative correlation between NO concentration and sperm concentration, the percentage of rapid progressive motility and arginase activity (r = -0.622, P < 0.01; r = -0.425, P < 0.05 and r = -0.445, P < 0.05, respectively). These results indicate that psychological stress causes an increase of NO level and a decrease of arginase activity in the L-arginine-NO pathway. Furthermore, poor sperm quality may be due to excessive production of NO under psychological stress. In the light of these results, we suggest that the arginine-NO pathway, together with arginase and NO synthase, are involved in semen quality under stress conditions.

Effect of psychological stress on the L-arginine-nitric oxide pathway and semen quality

It has been reported that mental stress causes abnormality of spermiogram parameters. We investigated the effect of psychological stress on the L-arginine-nitric oxide (NO) pathway. Semen samples were collected from 29 healthy fourth semester medical students just before (stress) and 3 months after (non-stress) the final examinations. Psychological stress was measured by the State Anxiety Inventory questionnaire. After standard semen analysis, arginase activity and NO concentration were measured spectrophotometrically in the seminal plasma. Measurements were made in duplicate. During the stress period, sperm concentration (41.28 ± 3.70 vs 77.62 ± 7.13 x 10(6)/mL), rapid progressive motility of spermatozoa (8.79 ± 1.66 vs 20.86 ± 1.63 percent) and seminal plasma arginase activity (0.12 ± 0.01 vs 0.22 ± 0.01 U/mL) were significantly lower than in the non-stress situation, whereas seminal plasma NO (17.28 ± 0.56 vs 10.02 ± 0.49 æmol/L) was higher compared to the non-stress period (P < 0.001 for all). During stress there was a negative correlation between NO concentration and sperm concentration, the percentage of rapid progressive motility and arginase activity (r = -0.622, P < 0.01; r = -0.425, P < 0.05 and r = -0.445, P < 0.05, respectively). These results indicate that psychological stress causes an increase of NO level and a decrease of arginase activity in the L-arginine-NO pathway. Furthermore, poor sperm quality may be due to excessive production of NO under psychological stress. In the light of these results, we suggest that the arginine-NO pathway, together with arginase and NO synthase, are involved in semen quality under stress conditions.

The effects of taking chronic nitrate by drinking water on thyroid functions and morphology.

Nitrate incorporation in humans takes place via drinking water and food. The water used for drinking and cooking in the goitrous areas is high in nitrate content. The aim of the present study was to evaluate both chronic effects and the dose-response relationship of nitrate on thyroid functions. A total of rats were divided into 5 work groups and sodium nitrate was added to their drinking water in different concentrations (0, 50, 100, 250 and 500 mg/l) over a 30-week period. The radioiodine uptake of thyroid was decreased in the 50 mg/l nitrate group, whereas it was increased in the 250 and 500 mg/l nitrate groups as compared to control. All hormones of thyroid gland except total thyroxine and thyroid-stimulating hormone were decreased in the 50, 250 and 500 mg/l nitrate groups. However, the level of total thyroxin was increased in the 100 mg/l nitrate group. Thyroid gland weights were increased in all experimental groups. Histomorphological changes were observed in the 250 and 500 mg/l nitrate groups. These findings suggest that nitrate impairs thyroid function involving the hypothalamo-hypophysio-thyroid axis. This observation could contribute to the current discussion about the acceptable daily intake of nitrate, as well as drinking water nitrate standard safety margins.

Glutathione and free sulphydryl content of seminal plasma in healthy medical students during and after exam stress.

BACKGROUND: It has been reported that there is a relationship between stress and infertility. The mechanisms of stress-related semen quality alterations have not been fully elucidated. In the present study, we investigated the effect of examination stress on seminal glutathione and free sulphydryl content and sperm quality. METHODS: Semen samples were collected from 34 healthy volunteers who were students of medical school in the fourth semester just before (stress period) and 3 months after (non-stress period) their final examinations. Their psychological examination stress was measured by the State Trait Anxiety Inventory (STAI) questionnaire. After standard semen analysis, semen samples were centrifuged at 10 000g for 15 min. Glutathione and free sulphydryl concentration of seminal plasma were measured. RESULTS: During the period of examination stress, the glutathione and free sulphydryl content of seminal plasma and the motility index of spermatozoa were significantly lower, whereas the percentage of morphologically abnormal spermatozoa was higher, than during the non-stress period (P < 0.001, for all). An association between seminal plasma glutathione and motility index was observed at both periods (P < 0.05 and P < 0.01, respectively). CONCLUSIONS: This study demonstrated that glutathione and free sulphydryl levels in seminal plasma decreased in subjects undergoing examination stress. Furthermore, poor sperm quality may be due to loss of glutathione and free sulphydryl content of seminal plasma.