Ökkeş Yilmaz

Melatonin suppresses cisplatin-induced nephrotoxicity via activation of Nrf-2/HO-1 pathway.

BackgroundCisplatin, one of the most effective and potent anticancer drugs, is used in the treatment of a wide variety of both pediatric and adult malignancies. However, the chemotherapeutic use of cisplatin is limited by its serious side-effects such as nephrotoxicity and ototoxicity. Cisplatin chemotherapy induces a reduction in the antioxidant status, leading to a failure of the antioxidant defense against free-radical damage generated by antitumor drugs. Cisplatin-induced oxidative stress in the kidney was partially prevented by antioxidant treatments using superoxide dismutase, glutathione, selenium and flavonoids. Melatonin and its metabolites possess free-radical scavenging activity and it has been shown that they protect against cisplatin toxicity. However, the mechanism of the protective effects of melatonin against cisplatin-induced nephrotoxicity is still essentially unknown. We therefore designed this study to investigate the underlying mechanism of the protective effect of melatonin against cisplatin-induced renal damage in a rat nephrotoxicity model in vivo.MethodsTwenty eight 8-week-old male Wistar rats were divided into four groups of control, melatonin treatment (4 mg/kg b.w i.p. for 10 days), cisplatin treatment (7 mg/kg b.w., i.p.) and melatonin and cisplatin combination treatment. Serum urea nitrogen (urea-N) and creatinine levels were measured. Histopathological changes were evaluated. In addition, we analyzed the expression levels of HO-1, Nrf2, NF-κB and AP-1 in Western blot analysis.ResultsBoth serum creatinine and urea nitrogen increased significantly following cisplatin administration alone; these values decreased significantly with melatonin co-treatment of cisplatin-treated rats. Histological analysis showed that cisplatin caused damage in the proximal tubular cells in the kidneys of cisplatin-treated rats; these changes were reversed by melatonin co-treatment. Upon Western blot analysis, melatonin treatment increased Nrf2 accumulation in the nuclear fraction, and increased the expression of HO-1 in the cytosolic fraction as compared to the cisplatin-treated rats. Expressions of NF-κB p65 and AP-1 were increased significantly in the kidneys of rats treated with cisplatin compared with the expression in the kidneys from the control, melatonin-only-treated and melatonin co-treated rats.ConclusionOur present data suggest that melatonin attenuates cisplatin-induced nephrotoxicity possibly by modulating Nrf2/HO-1 signaling.

Effect of vitamin C and lipoic acid on streptozotocin-induced diabetes gene expression: mRNA and protein expressions of Cu–Zn SOD and catalase

The involvement of oxidative stress in the pathogenesis of diabetes mellitus has been confirmed by numerous studies. In this study, the expression of two antioxidant enzymes, superoxide dismutase (SOD), and catalase which are involved in the detoxification of reactive oxygen species was studied in the streptozotocin-induced diabetic rat liver tissues. The enzyme assays showed a significant decrease in both enzymes activities compared to control animals. The RT-PCR and Western-blot analysis results demonstrated that this decrease in activity is regulated at the level of gene expression, as both catalase and Cu–Zn SOD mRNA and protein expressions were also suppressed. Supplementing the animals with vitamin C, a powerful antioxidant increased both SOD and catalase activities with no change in both mRNA and protein expressions suggesting a role of post-translational modification. However, even though mRNA expressions of both catalase and Cu–Zn SOD were not changed, the protein levels increased in parallel to activities in the case of another antioxidant, α-lipoic acid. An increase in the rate of translation, without changing the rate of transcription indicates a translational effect of lipoic acid in changing the activities of antioxidant enzymes to prevent the oxidative damage in diabetes.

Protective effects of nanostructures of hydrated C60 fullerene on reproductive function in streptozotocin-diabetic male rats

Diabetes mellitus is a well-recognized cause of male sexual dysfunction and impairments of male fertility. Streptozotocin (STZ) is used for medical treatment of neoplastic islet β-cells of pancreas and producing of animal model of diabetes mellitus type 1 that is characterized by suppression of reproductive activity due to the hyperglycaemia-induced oxidative stress and histopathological alterations in testes. Seeking for the agents that could alleviate diabetes-induced damage to reproductive system is yet the important area of inquiry. The present study was designed to evaluate whether hydrated C(60) fullerene (C(60)HyFn), which is known to be powerful bioantioxidant, eliminate testicular dysfunction induced by STZ-diabetes in rats. Wistar strain male albino rats were divided into four groups of six animals each: (1) control group, (2) C(60)HyFn-treated nondiabetic group, (3) STZ-diabetic group and (4) C(60)HyFn-treated diabetic group. Once hyperglycaemia was induced by STZ, rats in the second and fourth groups were treated with C(60)HyFn (in the form of drinking water) at the dose of 4μg/kg daily for 5 weeks. In diabetic rats, relative weights of right cauda epididymis, seminal vesicles, prostate, sperm motility and epididymal sperm concentration were significantly less than those of control group, but which were restored in the fourth group treated with C(60)HyFn (p<0.001). In hematoxylin and eosin staining, marked histopathological changes including degeneration, desquamation, disorganisation and reduction in germinal cells, interstitial oedema and congestion were evident in the testis of diabetic rats, but C(60)HyFn treatment resulted in recovery of histopathological changes and an increase in Johnsens testicular score significantly (p<0.001). C(60)HyFn treatment restores the increased apoptosis induced by STZ-diabetes. In diabetic rats, levels of serum testosterone, testicular reduced glutathione (GSH) and alpha-tocopherol were significantly reduced and testicular lipid peroxidation level was increased (p<0.001). Nevertheless, treatment of diabetic rats with C(60)HyFn resulted in significant corrective effects on these parameters towards the control levels. C(60)HyFn, applied alone, did not exert any toxic effects in testicular tissues. Furthermore, C(60)HyFn treatment in diabetic and nondiabetic rats resulted in considerable elevations of some important polyunsaturated fatty acids. In conclusion, we have presented for the first time substantial evidence that administration of C(60)HyFn significantly reduces diabetes-induced oxidative stress and associated complications such as testicular dysfunction and spermatogenic disruption.

Insecticide imidacloprid induces morphological and DNA damage through oxidative toxicity on the reproductive organs of developing male rats

We investigated whether treatment with imidacloprid would induce morphological changes, DNA fragmentation, antioxidant imbalance and apoptosis in the reproductive system of developing male rats. Twenty‐four male rats were included in this 90‐day study, starting at 7 days of age. The rats were divided into four groups. The first group was used as control. The second, third and fourth groups received oral 0.5‐, 2‐ and 8‐mg/kg imidacloprid, respectively. Serum, sperm and testis samples were collected from all groups at the end of the experimental period. The weights of the epididymis, vesicula seminalis, epididymal sperm concentration, body weight gain, testosterone and reduced glutathione values were lower in the imidacloprid‐treated groups than that in the controls. All treated groups had increased lipid peroxidation, fatty acid concentrations and higher rates of abnormal sperm. Apoptosis and fragmentation of seminal DNA were higher in rats treated at the two higher doses of imidacloprid. These results show that this compound has a negative effect on sperm and testis of rats. Copyright

Effects of Certain Micronutrients and Melatonin on Plasma Lipid, Lipid Peroxidation, and Homocysteine Levels in Rats

BACKGROUND Numerous studies suggest an association between high intake of antioxidant vitamins and fish oil and reduced risk of coronary heart disease. Hyperhomocysteinemia has also been identified as an independent risk factor for arteriosclerosis. In this paper, we aimed to evaluate the effects of vitamin E, vitamin C, vitamin C 6 palmitate (VC6P), lipoic acid, fish oil, and melatonin supplementation on lipid peroxidation, plasma lipid, and homocysteine (Hcy) levels in rats. METHODS Animals were divided into seven groups: one was used as control and each remaining group was supplemented with one substance for 6 weeks. All substances were dissolved in olive oil and injected intraperitoneally (i.p.) with the exception of vitamin C, which was dissolved in drinking water. Plasma Hcy, lipid peroxidation, and lipids were determined. RESULTS Plasma malondialdehyde (MDA) levels decreased significantly in melatonin (p <0.01), lipoic acid (p <0.01), and vitamin E (p <0.05) groups. On the other hand, supplementation with vitamin C and VC6OP lowered MDA levels moderately but not significantly (p >0.05). Fish oil supplementation caused a slight but insignificant increase in plasma MDA levels (p >0.05). Plasma lipid levels in animals treated with melatonin, vitamin E, vitamin C, lipoic acid, and fish oil were significantly lower than those of controls; however, treatment of rats with VC6P has no significant effect on plasma lipid level. Melatonin and fish oil administration significantly lowered plasma Hcy levels, whereas VC6P elevated its level. There was no significant effect of vitamin E, vitamin C, and lipoic acid on levels of plasma Hcy. CONCLUSIONS Our data suggest that supplementation with antioxidants appears to be hypolipidemic. In addition to these beneficial effects, administration of melatonin and fish oil deserves careful consideration as a measure to lower plasma Hcy levels and reduce risk of cardiovascular diseases.

Effects of triple antioxidant combination (vitamin E, vitamin C and α-lipoic acid) with insulin on lipid and cholesterol levels and fatty acid composition of brain tissue in experimental diabetic and non-diabetic rats

The aim of this research was to examine the effects of a triple antioxidant combination (vitamins E (VE) and C (VC) plus α‐lipoic acid (LA)) on the total lipid and cholesterol levels and the fatty acid composition of brain tissues in experimental diabetic and non‐diabetic rats. VE and LA were injected intraperitoneally (50 mg/kg) four times per week and VC was provided as a supplement dissolved in the drinking water (50 mg/kg). In addition, rats in the diabetes 1 and D + VELAVC groups were given daily by subcutaneous insulin injections (8 IU/kg), but no insulin was given to rats in the diabetes 2 group.

Effects of alpha lipoic acid, ascorbic acid-6-palmitate, and fish oil on the glutathione, malonaldehyde, and fatty acids levels in erythrocytes of streptozotocin induced diabetic male rats

In this research, it has been aimed to evaluate the improvement effects of alpha lipoic acid (ALA), ascorbic acid‐6‐palmitate (AA6P), fish oil (FO), and their combination (COM) on some biochemical properties in erythrocytes of streptozotocin (STZ)‐induced diabetic male rats. According to experimental results, glutathione (GSH) level in erythrocytes decreased in diabetes (P < 0.01), D + ALA, and D + AA6P groups (P < 0.001). Malonaldehyde (MA) level increased in diabetes (P < 0.05), D + FO, and D + COM groups (P < 0.001), but its level in D + AA6P and D + ALA groups was lower in diabetes group (P < 0.01). Total lipid level in diabetes and diabetes plus antioxidant administered groups were higher than control. Total cholesterol level was high in diabetes and D + ALA groups (P < 0.05), but its level reduced in D + FO compared to control and diabetes groups, P < 0.05, < 0.001, respectively. Total triglyceride (TTG) level was high in the D + ALA (P < 0.05) and D + COM (P < 0.001) groups. In contrast, TTG level in blood of diabetes group was higher than diabetes plus antioxidant and FO administered groups (P < 0.001). According to gas chromatography analysis results, while the palmitic acid raised in diabetes group (P < 0.05), stearic acid in D + FO, D + ALA, and diabetes groups was lower than control (P < 0.05), oleic acid reduced in D + COM and D + FO groups, but its level raised in D + AA6P and D + ALA groups (P < 0.01). As the linoleic acid (LA) elevated in ALA + D, D + AA6P, and diabetes groups, linolenic acid level in diabetes, D + AA6P, and D + FO groups was lower than control (P < 0.001). Arachidonic acid (AA) decreased in D + ALA, D+ AA6P, and diabetes groups (P < 0.01), but its level in D + COM and D + FO was higher than control (P < 0.05). Docosahexaenoic acid (DHA) increased in D + AA6P and D + COM (P < 0.05). While the total saturated fatty acid level raised in diabetes group, its level reduced in D + ALA and D + FO groups (P < 0.05). In contrast, total unsaturated fatty acid level in D + ALA and D + FO groups was higher than control (P < 0.05). In conclusion, present data have confirmed that the combination of the ALA, AA6P, and FO have improvement effects on the recycling of GSSG to reduced GSH in erythrocytes of diabetic rats, and in addition to this, oxidative stress was suppressed by ALA and AA6P, and unsaturated fatty acid degree was raised by the effects of ALA and FO. J. Cell. Biochem. 86: 530–539, 2002.

Resveratrol (trans-3,4′,5-trihydroxystilbene) decreases lipid peroxidation level and protects antioxidant capacity in sera and erythrocytes of old female Wistar rats induced by the kidney carcinogen potassium bromate

Resveratrol is a natural polyphenolic compound and it is found in number of edible plants, especially grapes and peanuts, has been shown to have anti-inflammatory, anti-oxidant, anti-tumor and anti-platelet activities. The aim of the study was to examine the effects of resveratrol on the level of malondialdehyde (MDA), homocysteine, cholesterol, GSH, GSSG and lipophylic vitamins in serum and erythrocytes of old female Wistar rats induced by the kidney carcinogen potassium bromate (KBrO(3)). In the study, total 30-old female Wistar rats were used, and the rats were randomly divided into three groups. The first group was used as a control, the second group KBrO(3) group, and third group R+KBrO(3). Rats in KBrO(3) and R+KBrO(3) groups were injected intraperitoneally a single dose KBrO(3) (80mg/kg) in physiologic saline buffer. After 2days, those in R+KBrO(3) group were intraperitoneally injected with resveratrol (33mg/kg) four times per week, and physiological saline was injected to control group rats. All the analysis was performed fully automatic with high performance liquid chromatography equipment. The results indicate that serum cholesterol level in the R+KBrO(3) group was higher than the control group (p<0.05), and level of the cholesterol in erythrocytes membranes was lower in the same group (p<0.01). The MDA level in serum and erythrocytes of the R+KBrO(3) were lower than the control and KBrO(3) groups (p<0.01). However, the MDA level in erythrocytes of the KBrO(3) group was high compared to the control group (p<0.05). Homocysteine and δ-tocopherol levels in serum of the R+KBrO(3) group were lower than the control group (p<0.05, p<0.001). α-Tocopherol level in serum and erythrocytes of the KBrO(3) group was lower than the control group (p<0.05), whereas its level was not found to differ between the control and R+KBrO(3) groups. GSH and GSSG levels in the KBrO(3) group of erythrocytes were higher than control group (p<0.05, p<0.01), however the ratio GSH/GSSG in the same group was lower than control group. In conclusion, our results confirm that the lipid peroxidation formation in serum and erythrocytes of old female Wistar rats by induced the KBrO(3) is prevented by the resveratrol. It was observed that the antioxidant capacity of erythrocytes was protected by resveratrol.

Assessment of imidacloprid toxicity on reproductive organ system of adult male rats.

In the current study it was aimed to investigate the toxicity of low doses of imidacloprid (IMI) on the reproductive organ systems of adult male rats. The treatment groups received 0.5 (IMI-0.5), 2 (IMI-2) or 8 mg IMI/kg body weight by oral gavage (IMI-8) for three months. The deterioration in sperm motility in IMI-8 group and epidydimal sperm concentration in IMI-2 and IMI-8 groups and abnormality in sperm morphology in IMI-8 were significant. The levels of testosterone (T) and GSH decreased significantly in group IMI-8 compared to the control group. Upon treatment with IMI, apoptotic index increased significantly only in germ cells of the seminiferous tubules of IMI-8 group when compared to control. Fragmentation was striking in the seminal DNA from the IMI-8 group, but it was much less obvious in the IMI-2 one. IMI exposure resulted in elevation of all fatty acids analyzed, but the increases were significant only in stearic, oleic, linoleic and arachidonic acids. The ratios of 20:4/20:3 and 20:4/18:2 were decreased and 16:1n-9/16:0 ratio was increased. In conclusion, the present animal experiments revealed that the treatment with IMI at NOAEL dose-levels caused deterioration in sperm parameters, decreased T level, increased apoptosis of germ cells, seminal DNA fragmentation, the depletion of antioxidants and change in disturbance of fatty acid composition. All these changes indicate the suppression of testicular function.

Antimicrobial and biological effects of ipemphos and amphos on bacterial and yeast strains

In this study, the antimicrobial effects of monophosphazenes such as SM ipemphos and amphos were examined on bacterial and yeast strains. In addition, the biological effects of these compounds were tested on the lipid level of Saccharomyces cerevisiae and Candida albicans cells. The SM has an antimicrobial effect on the bacterial and yeast strains within the range of 100 and 1500 μg. When the concentration was increased, the inhibition zone expanded on the growth media ( p < 0·01; p < 0·001). The ipemphos did not affect the bacterial and yeast cells in the 100 and 600 μg range. In addition, the amphos did not show an antimicrobial effect on the bacterial cells between 100 and 300 μg or on yeast cells at any of the administered concentrations. In vitro media, the biological effects of these molecules were compared with vitamin E, melatonin and fish oil on the yeast cells. We have found that monophosphazenes have growth effects on the cells in vitro media. The lipid level of S. cerevisiae cells was decreased by 300 μg doses of vitamin E, fish oil, and ipemphos (respectively; p < 0·05, p < 0·01, and p < 0·001). In addition, the lipid levels of the same yeast cells were depressed by 1000‐μg doses in all supplemented groups. However, it was observed that the highest decrease in lipid level of S. cerevisiae cells occurred in the amphos group ( p < 0·001). The lipid levels of the C. albicans cells were significantly reduced ( p < 0·01) by 300 μg of amphos and melatonin. In contrast, the vitamin E and fish oil significantly raised ( p < 0·01; p < 0·001) the lipid level of the same yeast cell, as compared with the control. In addition, the lipid level of these cells was increased by administration of 1000 μg vitamin E, and melatonin ( p < 0·01). In conclusion, while high concentrations of ipemphos and amphos have an antimicrobial effect on bacterial and yeast cells, amphos did not affect the yeast cells. While ipemphos and amphos increased cell growth in media, they reduced the lipid level of C. albicans and S. cerevisiae. In addition, the antioxidants such as vitamin E, melatonin, and fish oils affected the lipid level of yeast cells. Copyright