Reduction of cryopreservation-induced structural, functional and molecular damages in ram sperm by hydrated C60 fullerene.

This study aimed to investigate the morphological, functional and molecular changes in frozen-thawed ram sperm using an extender containing different concentrations of hydrated carbon 60 fullerene (C60 HyFn), a nanotechnological product. Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3 weeks. Each pooled semen sample was divided into six equal groups and diluted with tris + egg yolk extender including 0 (control), 200, 400, 800 nM, 1 and 5 µM concentrations of C60 HyFn at 37°C. They were then frozen in liquid nitrogen vapour at -140°C, stored in liquid nitrogen container (-196°C) and thawed at 37°C for 25 s before analysis. In comparison with control, C60 HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase-3 and DNA fragmentation levels in frozen-thawed ram semen. When compared to control, C60 HyFn supplementation significantly down-regulated the expression levels of miR-200a and KCNJ11, and significantly up-regulated the expression levels of miR-3958-3p (at the concentrations of 200, 400, 800 nM and 1 µM), CatSper1 (at the concentrations of 200, 400 nM and 5 µM), CatSper2 (at the concentrations of 1 and 5 µM), CatSper3 (at the concentrations of 200, 400 nM, 1 and 5 µM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800 nM, 1 and 5 µM) and TRPV5 (at the concentrations of 200, 400 and 800 nM). The addition of C60 HyFn had no effect on global DNA methylation rates. As a result, C60 HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze-thawing procedure.

Effect of hydrated C60 fullerene on lipid, vitamin and amino acid composition in frozen-thawed ram semen.

This study was conducted to investigate the effect of different doses of hydrated C60 fullerene (C60HyFn) on freeze-thawing process-induced changes in lipid, vitamin and amino acid composition and also in motility, kinematic, sperm quality and oxidative stress parameters in ram semen. Semen was collected from seven rams twice a week for 3 weeks, so six repetitions were performed. The semen collected in each repetition was pooled. Each pooled sample was diluted with tris + egg yolk extender with (200 nM, 400 nM, 800 nM, 1 µM and 5 µM) and without (control) C60HyFn and they were frozen in mini straws. The doses of 800 nM, 1 µM and 5 µM had higher total, progressive motility, sperm membrane functionality rates, glutathione-peroxidase and catalase activities. All doses of C60HyFn significantly reduced dead and total abnormal sperm rates and malondialdehyde levels. Significant increases in vitamin A (400 and 800 nM doses), vitamin K1 (400 nM, 800 nM and 1 µM doses), total amino acid (all doses) levels, but significant decreases in vitamin D2 (800 nM, 1 and 5 µM doses), vitamin D3 (1 and 5 µM doses) and vitamin E (200 nM, 1 and 5 µM) levels were observed compared to control. In conclusion, the addition of C60HyFn to ram semen at 200 nM - 5 µM range, especially at a dose of 800 nM, provides a positive contribution to the protection of motility, vitamins A, K and total amino acid levels, and oxidant/antioxidant balance after freeze-thawing.

Potential ameliorative effect of dietary quercetin against lead-induced oxidative stress, biochemical changes, and apoptosis in laying Japanese quails.

Lead (Pb) is a widespread environmental pollutant which is a toxic threat to human and animal health. The present study was designed to evaluate the ameliorative role of quercetin in laying quails exposed to Pb. A total of 112 birds were randomly divided into four groups. The control group was fed with basal diet, the Pb group was fed with ration supplemented with Pb at the dose of 100 mg/kg (as Pb (II) acetate trihydrate), the Quercetin group was fed with ration supplemented with quercetin at the dose of 400 mg/kg, and the Pb+ Quercetin group was fed with ration supplemented with Pb at the dose of 100 mg/kg and quercetin at dose of 400 mg/kg. Results showed that serum total protein, glucose, albumin, and blood urea nitrogen (BUN) values of the Pb + Quercetin group partially improved with quercetin supplementation. Meanwhile, serum creatinine values of the Pb + Quercetin group was found to be significantly lower than that of the Pb group. Aspartate aminotransferase (AST) and alanine transaminase (ALT) enzyme activities in the Quercetin and Pb + Quercetin groups were similar to those of the Control group, unlike the Pb group. Moreover, alkaline phosphatase (ALP) enzyme activity of the Pb + Quercetin group significantly improved with the addition of quercetin. We also found that malondialdehyde (MDA) levels of the kidney, liver, and heart were significantly reduced by quercetin supplementation. The glutathione, catalase, and glutathione peroxidase activities of the kidney, liver, and heart tissue were increased by quercetin supplementation. These results were in line with the observed apoptotic markers. The expression of caspase-3 and caspase-9 were significantly decreased by quercetin supplementation. It may be concluded that dietary supplementation with quercetin ameliorates the toxic effects of Pb exposure by alleviating oxidative stress, biochemical changes, and apoptosis in quails.

Effect of Hydrated Carbon 60 Fullerene on Frozen Ram Semen Quality.

The aim of this study was to evaluate the effect of hydrated carbon 60 fullerene (C60HyFn) on ram semen quality during cryopreservation. Three ejaculates from each of seven Akkaraman rams were collected using an artificial vagina during the nonbreeding season and pooled. Pooled semen samples were divided into 10 equal parts and diluted with tris + egg yolk extender not containing (control) and containing 100, 200, 400, and 800 nM and 1, 5, 10, 20, and 40 µM C60HyFn at 37°C. After addition of 5% glycerol and an equilibration process for 3 hours, the samples were frozen in 0.25-mL straws in an automatic freezing device at -140°C and stored in a liquid nitrogen container. Straws were thawed 24 hours after freezing and analyzed immediately with no incubation period. Motility, kinematic parameters, abnormality, vitality, hypo-osmotic swelling test (HOST), and oxidative stress levels were analyzed in thawed semen. Compared with the control, 200, 400, and 800 nM and 1 and 5 µM C60HyFn doses increased motility and HOST values and decreased the dead sperm rate. When compared with the control, addition of C60HyFn significantly decreased malondialdehyde levels (between 200 nM and 40 µM doses) and significantly increased glutathione peroxidase (between 800 nM and 40 µM doses) and catalase (between 1 and 40 µM doses) activities. In conclusion, results of this study show that the C60HyFn nanoparticles are nontoxic to ram semen and their supplementation in the extender is beneficial to sperm motility and membrane integrity after freeze-thawing.

Changes in heavy metal levels, reproductive characteristics, oxidative stress markers and testicular apoptosis in rams raised around thermal power plant.

Male reproductive dysfunction is one of the damages in the organism caused by heavy metals. In this study, it was aimed to investigate the changes in heavy metal levels in serum and testicular tissue, and serum hormone levels, epididymal spermatozoa characteristics, tissue oxidative stress levels, testicular histology and testicular apoptosis level in rams raised in remote and near regions of a thermal power plant. A total of 75 rams were divided into 3 groups according to the regions, where they were born and raised, being far [250 km distance, group 1 (control), n = 25], close (20 km distance, group 2, n = 25) and very close (10 km distance, group 3, n = 25) to the thermal power plant. The blood along with testis and epididymis tissues was taken from the animals after slaughtering. In addition, soil and water heavy metal analyzes were also performed. The highest levels of serum Al, Cr, As, Ag, Sn and testicular Al, V, Ni, Ag, Cd, Cr, As, Pb, and the lowest levels of serum Cu, testicular Cu and Zn were determined in group 3 compared to control. Soil and water heavy metal results were similar to those found in serum and testis. The lowest serum testosterone level, tissue glutathione-peroxidase and catalase activities, spermatozoon concentration, and the highest tissue malondialdehyde level, dead spermatozoon rate, Bax apoptotic protein expression level and Bax/Bcl-2 rate alongside some testicular histopathological lesions were found in group 3 in comparison to control. Significant correlations were determined between some heavy metal levels and some parameters measured. As a result, heavy metals accumulate in the soil and water in the region close to the thermal power plant. The endocrine and exocrine reproductive potentials of rams born and grown in these regions were clearly damaged by the increased testicular heavy metals due to water drank and herbs consumed.

Effect of freeze-thawing process on lipid peroxidation, miRNAs, ion channels, apoptosis and global DNA methylation in ram spermatozoa.

This study was carried out to investigate the effect of the semen freeze-thawing process on the functionality and molecular structure of ram spermatozoa. The temperature of pooled and diluted semen at 38°C (group 1, control) was lowered to 5°C (group 2), and it was subjected to glycerolisation-equilibration (group 3), frozen and thawed (group 4). Compared to the control, deterioration in spermatological parameters and significant increases in lipid peroxidation and global DNA methylation levels were observed in groups 3 and 4. When compared with the control, significant downregulation in the levels of miR-485 of group 2, miR-29a of group 3 and let-7a, miR-485 and miR-29a of group 4, and significant upregulation in the levels of miR-107 of group 3 and miR-127 of groups 3 and 4 were detected. In comparison to the control, significant upregulation in the levels of CatSper1, CatSper2, CatSper3, CatSper4, ANO1 and TRPM3 of group 2, CatSper4, ANO1 and TRPM3 of group 3 and KCNJ11 of group 4, and significant downregulation in the CatSper 3 level of group 4 were determined. As a result, the semen freeze-thawing process causes motility and morphological disorders in rams. This may be due to molecular changes associated with lipid peroxidation in spermatozoa.

The in-vivo assessment of Turkish propolis and its nano form on testicular damage induced by cisplatin.

OBJECTIVE: Chemotherapeutic drugs, such as cisplatin (CP), which are associated with oxidative stress and apoptosis, may adversely affect the reproductive system. This study tests whether administration of propolis and nano-propolis (NP) can alleviate oxidative stress and apoptosis in rats with testicular damage induced by CP. METHODS: In this study, polymeric nanoparticles including propolis were synthesized with a green sonication method and characterized using Fourier transform-infrared spectroscopy, Brunauer-Emmett-Teller, and wet scanning transmission electron microscopy techniques. In total, 56 rats were divided into the following seven groups: control, CP, propolis, NP-10, CP + propolis, CP + NP-10, and CP + NP-30. Propolis (100 mg/kg), NP-10 (10 mg/kg), and NP-30 (30 mg/kg) treatments were administered by gavage daily for 21 d, and CP (3 mg/kg) was administered intraperitoneally in a single dose. After the experiment, oxidative stress parameters, namely, malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), and catalase (CAT), and apoptotic pathways including B cell leukemia/lymphoma-2 protein (Bcl-2) and Bcl-2-associated X protein (Bax) were measured in testicular tissues. Furthermore, sperm quality and weights of the testis, epididymis, right cauda epididymis, seminal vesicles and prostate were evaluated. RESULTS: Propolis and NP (especially NP-30) were able to preserve oxidative balance (decreased MDA levels and increased GSH, CAT, and GPx activities) and activate apoptotic pathways (decreased Bax and increased Bcl-2) in the testes of CP-treated rats. Sperm motility in the control, CP, and CP + NP-30 groups were 60%, 48.75%, and 78%, respectively (P < 0.001). Especially, NP-30 application completely corrected the deterioration in sperm features induced by CP. CONCLUSION: The results show that propolis and NP treatments mitigated the side effects of CP on spermatogenic activity, antioxidant situation, and apoptosis in rats.

Protective role of yeast beta-glucan on lead acetate-induced hepatic and reproductive toxicity in rats.

Lead (Pb) is one of the most common environmental pollutants and causes adverse effects on human and animal health. This study aimed to evaluate the protective role of beta-glucan against hepatic and reproductive toxicity induced by lead acetate. A total of 28 Sprague Dawley male rats were distributed into four groups (n = 7). The control group was intraperitoneally injected saline (1 ml/kg b.w.) daily for 21 days, the Pb group was intraperitoneally injected lead acetate (15 mg/kg b.w.) daily for 21 days, the beta-glucan group was orally administrated beta-glucan (50 mg/kg b.w.) daily for 21 days, and the Pb + beta-glucan group was intraperitoneally injected lead acetate (15 mg/kg b.w.) daily following the oral administration of beta-glucan (50 mg/kg b.w.) daily for 21 days. Results showed that feed intake in the Pb + beta-glucan group was significantly increased in comparison with that of the Pb group (p < 0.001). We also found that liver malondialdehyde (MDA) level was increased significantly in the Pb group (p < 0.01), while glutathione (GSH) level (p < 0.05), glutathione peroxidase (GSH-Px) (p < 0.05), and catalase (CAT) (p < 0.01) activities were reduced when they were compared with control. Moreover, Pb administration increased expression of pro-apoptotic protein Bax, the ratio of Bax/Bcl-2, and decreased the expression of the antiapoptotic protein Bcl-2 (p < 0.01). Also, Pb was found to cause a significant decrease in sperm motility (p < 0.01) and sperm concentration (p < 0.05) but increase in sperm tails and total sperm anomalies (p < 0.05). These findings were partially preserved by the administration of beta-glucan. Taken together, these results indicated that beta-glucan has the potential to alleviate the Pb-induced toxicity.

Ellagic acid plays an important role in enhancing productive performance and alleviating oxidative stress, apoptosis in laying quail exposed to lead toxicity.

Lead (Pb) is one of the most toxic heavy metal environmental pollutants due to its widespread use of the industry and it is a harmful substance for human and animal health. This study was conducted to investigate the potential protective effects of ellagic acid (EA) on performance, egg quality, antioxidant parameters, and apoptotic pathway proteins in laying quails exposed to Pb toxicity. A total of 168 (15-week old) laying quails (Coturnix coturnix Japonica) were divided into 6 experimental groups (with similar initial average body weight). Birds were fed 1 of 6 diets for 8 weeks: 1 - Control (basal diet), 2 - Pb (basal diet + 100 mg/kg Pb), 3 - EA-300 (basal diet + 300 mg/kg EA), 4 - EA-500 (basal diet + 500 mg/kg EA), 5 - Pb + EA-300 (basal diet + 100 mg/kg Pb + 300 mg/kg EA), 6 - Pb + EA-500 (basal diet + 100 mg/kg Pb + 500 mg/kg EA). The results showed that adding 100 mg/kg of Pb to basal diet was adversely affected the performance parameters and, feed intake and egg production were significantly decreased by Pb supplementation (P < 0.01). However, the EA supplementation to Pb groups improved the performance parameters. Compared with the Pb alone group, in Pb + EA-500 group increased egg production by 8.4%. There were no significant differences in the Haugh unit, albumen index, and yolk index among groups (P > 0.05). Liver and kidney tissues of Pb group malondialdehyde (MDA) level increased (P < 0.001) and, GSH, GSH-Px, and CAT values decreased (P < 0.001) but, EA supplementation alleviated this condition (P < 0.001). The protein levels of caspase-3 and -9 were significantly increased in the Pb group compared to the control group, whereas EA supplementation alleviated the Pb-induced apoptosis by decreasing caspase-3 and -9 levels in the liver tissue (p < 0.001). In laying quails exposed to Pb toxicity, EA supplementation improves the performance parameters, enhances the antioxidant defense system, and suppresses apoptosis via regulates the expression of caspase-3 and -9. Thus, it was concluded that EA (especially 500 mg/kg) can ameliorate the toxic effects of Pb exposure in quails.

Protective effects of carvacrol against diabetes-induced reproductive damage in male rats: Modulation of Nrf2/HO-1 signalling pathway and inhibition of Nf-kB-mediated testicular apoptosis and inflammation.

Diabetes mellitus, which causes many complications, also adversely affects reproductive system in men. Studies reported that natural antioxidants are effective in reducing important complication risks caused by diabetes. Carvacrol is an antioxidant phenolic monoterpene compound with therapeutic effect in various diseases found in essential oils of aromatic plants such as pepper, wild bergamot and thyme. We aimed to investigate the effects of carvacrol on diabetes-induced reproductive damage in male rats by evaluating the Nrf2/HO-1 pathway and Nf-kB-mediated apoptosis/inflammation and spermatological parameters. For this purpose, 74 Wistar albino male rats were used. The diabetes model was performed using single-dose intraperitoneal injection of streptozotocin 55 mg/kg. Rats were fed with carvacrol 75 mg/kg/daily/gavage for 4 and 8 weeks. Rats were divided into four groups: control group, diabetic group, carvacrol group which fed with carvacrol and the diabetic group which fed with carvacrol. It was determined that carvacrol significantly decreased malondialdehyde levels, Bax,COX-2,Nf-kB protein expression levels, Bax/Bcl-2 ratio and significantly increased glutathione peroxidase, catalase activities, Bcl-2, Nrf2,HO-1 protein expression levels and it was determined that had a positive effect on spermatological parameters. In conclusion, the negative effects of diabetes in the male reproductive system can be prevented and/or reduced by giving carvacrol.

Turkish Propolis and Its Nano Form Can Ameliorate the Side Effects of Cisplatin, Which Is a Widely Used Drug in the Treatment of Cancer.

This study was performed to determine the effects of chitosan-coated nano-propolis (NP), which is synthesized via a green sonochemical method, and propolis on the side effects of cisplatin (CP), which is a widely used drug in the treatment of cancer. For this aim, 56 rats were divided into seven groups, balancing their body weights (BW). The study was designed as Control, CP (3 mg/kg BW at single dose of CP as intraperitoneal, ip), Propolis (100 mg/kg BW per day of propolis by gavage), NP-10 (10 mg/kg BW of NP per day by gavage), CP + Propolis (3 mg/kg BW of CP and 100 mg/kg BW of propolis), CP + NP-10 (3 mg/kg CP and 10 mg/kg BW of NP), and CP + NP-30 (3 mg/kg BW of CP and 30 mg/kg BW of NP). Propolis and NP (especially NP-30) were preserved via biochemical parameters, oxidative stress, and activation of apoptotic pathways (anti-apoptotic protein: Bcl-2 and pro-apoptotic protein: Bax) in liver and kidney tissues in the toxicity induced by CP. The NP were more effective than propolis at a dose of 30 mg/kg BW and had the potential to ameliorate CP's negative effects while overcoming serious side effects such as liver and kidney damage.

Nobiletin attenuates acetaminophen-induced hepatorenal toxicity in rats.

The study aimed to examine the effects of nobiletin on the toxicity model induced with acetaminophen (APAP). For this purpose, 24 adult male rats were equally divided into four groups. The groups were the control group (group 1); dimethyl sulfoxide only, the APAP group (group 2) received a single dose of APAP 1000 mg/kg on the 10th day of experiment; the Nobiletin group (group 3), nobiletin (10 mg/kg) for 10 days; and the APAP + Nobiletin group (group 4), nobiletin (10 mg/kg) for 10 days with a single dose of APAP (1000 mg/kg) administered on the 10th day and the experiment ended after 48 hours. At the end of the study, a significant increase in malondialdehyde, interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels and a significant decrease in glutathione levels, glutathione peroxidase activities and nuclear factor erythroid-derived 2-like 2 (Nrf-2) and heme oxygenase-1 (HO-1) expressions were observed with APAP application in liver and kidney tissues. Serum aspartate transaminase (AST), alanine transaminase (ALT), urea, and creatinine levels were also significantly increased in the APAP group. However, nobiletin treatment in group 4 reversed oxidative stress and inflammatory and histopathological signs caused by APAP. It is concluded that nobiletin may be a beneficial substance that confers hepatorenal protection to APAP-induced toxicity via antioxidant and anti-inflammatory mechanisms.

Tyrosol prevents AlCl3 induced male reproductive damage by suppressing apoptosis and activating the Nrf-2/HO-1 pathway.

Aluminium is a ubiquitous element that occurs naturally in the soil making human exposure to it is unavoidable. Tyrosol is present in olive oil and is known to have antioxidant effects. Therefore, the present study explores the toxic effects of aluminium chloride (AlCl3 ) and evaluates the possible protection by tyrosol in male rats. Testicular injury was induced by the administration of AlCl3 (34 mg kg-1  day-1 ). Rats were treated with either tyrosol (20 mg kg-1 day-1 ) or AlCl3 (34 mg kg-1 day-1 ). The experiment lasted for 10 weeks. Biochemical, histopathological and protein expression profiles were determined to decipher the role of tyrosol in protecting the cellular damage. Further, histomorphometric analyses of testes showed deranged architecture along with other noted abnormalities. AlCl3 group rats' testes showed decreased GSH levels, CAT activities, Nrf-2, HO-1, bcl-2 expressions and sperm motility whereas increased caspase-3 expressions, MDA levels, abnormal and dead/live sperm ratio. However, tyrosol treatment attenuated these changes. The present results demonstrate the beneficial role of tyrosol treatment in AlCl3 induced testicular toxicity alterations of rat.