Reproductive Toxicity In Male Mice Research Articles

Alleviative Effect of Rutin on Zearalenone-Induced Reproductive Toxicity in Male Mice by Preventing Spermatogenic Cell Apoptosis and Modulating Gene Expression in the Hypothalamic-Pituitary-Gonadal Axis.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin found in many agricultural products and can cause reproductive disorders, mainly affecting spermatogenesis in male animals. Rutin (RUT) is a natural flavonoid compound recognized for its significant antioxidant, anti-inflammatory and estrogenic properties. The present study aimed to determine the protective role of RUT against ZEN-induced reproductive toxicity in male mice. Twenty-four adult Kunming male mice were divided into four groups: control, RUT (500 mg/kg RUT), ZEN (10 mg/kg ZEN), ZEN + RUT (500 mg/kg RUT + 10 mg/kg ZEN), with six replicates per treatment. The results indicated that RUT mitigated ZEN-induced disruption in spermatogenic cell arrangement, decreased spermatozoa count, and increased sperm mortality in the testes. RUT significantly restored ZEN-induced reduction in T, FSH, LH, and E2 serum levels. Moreover, RUT mitigated ZEN-induced apoptosis by increasing the mRNA expression level of bcl-2, decreasing the mRNA expression level of kiss1-r, and decreasing the protein expression level of caspase 8 in reproductive tissues. These findings indicate the protective role of RUT against ZEN-induced reproductive toxicity in male mice by regulating gonadotropin and testosterone secretions to maintain normal spermatogenesis via the HPG axis, which may provide a new application direction for RUT as a therapeutic agent to mitigate ZEN-induced reproductive toxicity.

Carbon Tetrachloride-Induced Testicular Toxicity and Histopathological Alteration in Male Swiss Albino Mice

Recently there has been an increased association between toxic substances present in the environment and male infertility. Carbon tetrachloride (CCL4) is widely used as a chemical intermediate and as a feedstock in the production of chlorofluorocarbons. CCL4 is highly toxic to the liver, kidney, testicle, brain and other tissues. Therefore, the present study was designed to identify its effect on the reproductive system in adult male mice. Thirty adult male albino mice were divided into three equal groups (n=10): the first group control, the second group received 0.1m/100g body weight olive oil and the third group received 0.1ml/100g/body weight CCL4 intraperitoneally every alternate day for three weeks. The results showed that CCl4 causes a significant decrease in body weight, sperm motility, sperm count and testosterone level, while it leads to a significant increase in the number of abnormal sperm morphology. Additionally, CCL4 caused apparent alterations in the histological structure of the testes. In conclusion, CCl4-induced reproductive toxicity in male mice.

Microplastics cause reproductive toxicity in male mice through inducing apoptosis of spermatogenic cells via p53 signaling.

Studies on male reproductive toxicity of microplastics are still scarce and the precise mechanism is not distinct. C57BL/6 male mice were given oral gavage treatments treated with 5 μm (MPs) and 80 nm (NPs) polystyrene microplastics every day for 60 consecutive days in a row at dosages of 0, 10 and 40 mg/kg/d. The major damage of MPs and NPs were assessed by the assays in vivo and in vitro. Transcriptome sequencing was applied to screen the key involved pathways. In the 10 mg/kg/d NPs group, there was an increase in testicular organ coefficient, and in the 40 mg/kg/d MPs group, an increase in epididymal weight was observed. Vacuolization of spermatogenic cell layer, interstitial congestion, and germ cell apoptosis were found in the testes of MPs and NPs treatment mice at different dose groups. Higher apoptosis rate was observed in GC-2 cells after MPs and NPs treatment at different concentrations. Transcriptome analysis suggested that p53 pathway might be the key signal pathway of the cell apoptosis, and the expressions of p53 and other markers of cell apoptosis were indeed altered after exposure to MPs and NPs. MPs and NPs can cause reproductive toxicity in male mice through inducing apoptosis of spermatogenic cells via p53 signaling pathway, indicating MPs and NPs exposure be an unnegligible risk factor for reproductive health in male mice.

Pumpkin seed ethanolic extract protects against escitalopram-induced reproductive toxicity in male mice

Objective: To investigate the protective role of pumpkin seed ethanolic extract against escitalopram-induced reproductive toxicity in male mice.Methods: Swiss albino male mice were randomly divided into five groups with six mice in each group. Group I received normal water orally, Group II, III, IV and V received escitalopram oxalate (10 mg/kg), pumpkin seed extract (300 mg/kg) plus escitalopram oxalate (10 mg/kg), escitalopram oxalate (20 mg/kg), and pumpkin seed extract (300 mg/kg) plus escitalopram oxalate (20 mg/kg), respectively. All test doses were continuously administered orally once daily per animal body weight for 30 days and 60 days. Body weight and sexual organ weight were evaluated on day 31 and 61. Effects of pumpkin seed extract on sperm parameters, biochemical parameters and histology of testis were also investigated.Results: Escitalopram 10 or 20 mg/kg caused reproductive toxicity in male mice after 30 and 60 days of treatment. However, simultaneous administration of escitalopram oxalate (10 or 20 mg/kg) with pumpkin seed extract (300 mg/kg) attenuated escitalopram-induced testicular toxicity. Significant increase in the body weight and relative organ weight was observed. Sperm count, sperm motility and viability significantly increased (P<0.05). The histopathological alterations caused by escitalopram was also ameliorated.Conclusions: Ethanolic extract of pumpkin seeds (300 mg/kg body weight) protects again reproductive toxicity induced by escitalopram. Therefore, dietary intake of pumpkin seed extract might be useful for male patients who expose to antidepressant drug due to depression.

Glycine protects the male reproductive system against lead toxicity via alleviating oxidative stress, preventing sperm mitochondrial impairment, improving kinematics of sperm, and blunting the downregulation of enzymes involved in the steroidogenesis

Lead (Pb) is a highly toxic heavy metal widely dispersed in the environment because of human industrial activities. Many studies revealed that Pb could adversely affect several organs, including the male reproductive system. Pb-induced reproductive toxicity could lead to infertility. Thus, finding safe and clinically applicable protective agents against this complication is important. It has been found that oxidative stress plays a fundamental role in the pathogenesis of Pb-induced reprotoxicity. Glycine is the simplest amino acid with a wide range of pharmacological activities. It has been found that glycine could attenuate oxidative stress and mitochondrial impairment in various experimental models. The current study was designed to evaluate the role of glycine in Pb-induced reproductive toxicity in male mice. Male BALB/c mice received Pb (20 mg/kg/day; gavage; 35 consecutive days) and treated with glycine (250 and 500 mg/kg/day; gavage; 35 consecutive days). Then, reproductive system weight indices, biomarkers of oxidative stress in the testis and isolated sperm, sperm kinetic, sperm mitochondrial indices, and testis histopathological alterations were monitored. A significant change in testis, epididymis, and Vas deferens weight was evident in Pb-treated animals. Markers of oxidative stress were also significantly increased in the testis and isolated sperm of the Pb-treated group. A significant disruption in sperm kinetic was also evident when mice received Pb. Moreover, Pb exposure caused significant deterioration in sperm mitochondrial indices. Tubular injury, tubular desquamation, and decreased spermatogenic index were histopathological alterations detected in Pb-treated mice. It was found that glycine significantly blunted oxidative stress markers in testis and sperm, improved sperm mitochondrial parameters, causing considerable higher velocity-related indices (VSL, VCL, and VAP) and percentages of progressively motile sperm, and decreased testis histopathological changes in Pb-exposed animals. These data suggest glycine as a potential protective agent against Pb-induced reproductive toxicity. The effects of glycine on oxidative stress markers and mitochondrial function play a key role in its protective mechanism.

Hesperidin partly ameliorates the decabromodiphenyl ether-induced reproductive toxicity in pubertal mice.

Many materials use polybrominated diphenyl ethers (PBDEs) as flame retardants. As one of the most common congeners of PBDEs, decabromodiphenyl ether (PBDE-209) is reported to harm reproductive health. However, little is known research on attenuating the reproductive toxicity induced by PBDE-209. The present study aimed to investigate the effects of hesperidin against PBDE-209-induced reproductive toxicity in male mice. Pubertal male C57BL/6J mice were exposed to PBDE-209 groups (20, 100, 500mg/kg·bw) and hesperidin groups (100mg/kg·bw PBDE-209 + 100mg/kg·bw hesperidin) for 8weeks. The results showed that PBDE-209 increased the amount of abnormal morphological sperms and decreased the sex hormone levels. PBDE-209 induced the histopathological lesions of seminiferous tubules and blood-testis barrier in mice testis. Expressions of apoptosis-associated proteins and mRNA (Bax, Bcl-2, etc.) were altered by the PBDE-209 treatment. PBDE-209 prominently increased the malondialdehyde (MDA) levels, the biomarker of oxidative stress. Hesperidin treatment partly alleviated PBDE-209-induced histopathological lesions and apoptosis in mice testis. These findings suggested that hesperidin partly protects against PBDE-induced reproductive toxicity in pubertal mice. We conclude that more work needs to be done to explore the appropriate dosage of hesperidin or find other drugs to protect against the reproductive toxicity of PBDEs.

Effect of aqueous garlic ( Allium sativum ) extract against di‐(2‐ethylhexyl) phthalate induced reproductive toxicity in male mice

This study was designed to investigate testicular and male reproductive tract histopathologies and lipid profile against di-ethylhexyl phthalate (DEHP) exposure in mice and curative potentials of aqueous garlic (Allium sativum) extract. Four groups (n=10) were named and treated as follow (a) control (C): (normal feed and drinking water + 0.2ml corn oil); (b) aqueous garlic extract group (AGE): (500 mg/kg body weight of aqueous garlic extract); (c) DEHP group: (500 mg/kg body weight of DEHP, dissolved in corn oil; (d) AGE + DEHP group (500 mg/kg body weight garlic aqueous extract, and DEHP 500 mg/kg body weight dissolved in corn oil). The doses were given once daily through gavages for 28 days and on the 29th day, all the animals were euthanized through cervical dislocation and reproductive organs and blood samples were collected. The results showed that exposure to DEHP caused a significant effect on body weight, testicular weight, serum cholesterol, triglycerides, lipid profile, average cross-sectional area (ACSA) of the seminiferous tubule, ACSA of the lumen of seminiferous tubule, spermatogenic cells, Leydig's cells number, vas deferens diameter, lumen, muscular thickness, and epithelial cell height of vas deferens. This study revealed that exposure to DEHP can be injurious to male reproductive health and aqueous garlic extract can decrease the toxic effects of DEHP in male mice.

Phycobiliproteins extract from Spirulina protects against single-dose cadmium-induced reproductive toxicity in male mice.

Cadmium (Cd) is known for its many toxic effects on male population such as hypogonadism and fertility difficulties, which are oftenly associated with oxidative stress. As beneficial food, Spirulina(Sp) has been proved efficient against the heavy metal toxicity. This capacity can be associated with its phycobiliproteins (PBP). In this study, the capability of PBP and Sp to treat Cd-induced oxidative damage on the testes and spermatozoa was considered. CD-1 strain mice were orally treated with either Sp or PBP for 10 days prior to single-dose Cd challenge. Sperm quality determinations and testicle histology analysis were performed. Testosterone on serum was measured using enzyme-linked immunosorbent assay (ELISA). Oxidative damage was determined. Antioxidant enzyme activity was analyzed by measuring the activity of super oxide dismutase (SOD), catalase (Cat), and glutathione peroxidase (GpX). The motility and viability of sperm decrease with Cd and improve with PBP and Sp, as the acrosomal reaction (AR) is diminished by PBPs. Testosterone levels decrease due to Cd, and only Sp maintains elevated levels. Cd increases the production of malondialdehyde in the spermatozoa, but not in testes; this production of malondialdehyde in the spermatozoa decreases in the presence of PBP. ROS only decreases with Cd, FBP, and Sp at high concentrations. Advanced oxidative protein products (AOPP) decrease with Cd and PBPs. Cat and GpX increase their activity with Cd and are altered by FBP. Cd produces vascular alterations testes. Within the seminiferous tubule, it produces areas of necrosis and apoptosis, which improve with PBPs and Sp. PBPs have a strong antioxidant activity as they show protective properties against Cd oxidative-induced toxicity on testes and sperm.

Protective effect of resveratrol against bisphenol A-induced reproductive toxicity in male mice

We investigated the effect of resveratrol on reproductive toxicity induced by Bisphenol A (BPA) in male mice. Forty mice were randomly classified into five groups: Control; Bisphenol A; Bisphenol A + Resveratrol; Resveratrol and vehicle. Histological alterations, oxidative stress markers, and sperm parameters were examined. BPA caused an evident decrease in sperm motility, count, viability, and oxidative stress markers (p < 0.05). Also, the increase in malondialdehyde, tissue damage, and abnormal sperm observed in the BPA group (p < 0.05) while the treatment of resveratrol significantly increased the above disarrays (p < 0.05). Overall, we reported that resveratrol reduces negative effects caused by BPA in spermatogenesis.

The Role of Mitochondrial Impairment and Oxidative Stress in the Pathogenesis of Lithium-Induced Reproductive Toxicity in Male Mice.

Lithium (Li+) is prescribed against a wide range of neurological disorders. Besides its excellent therapeutic properties, there are several adverse effects associated with Li+. The impact of Li+ on renal function and diabetes insipidus is the most common adverse effect of this drug. On the other hand, infertility and decreased libido is another complication associated with Li+. It has been found that sperm indices of functionality, as well as libido, is significantly reduced in Li+-treated men. These adverse effects might lead to drug incompliance and the cessation of drug therapy. Hence, the main aims of the current study were to illustrate the mechanisms of adverse effects of Li+ on the testis tissue, spermatogenesis process, and hormonal changes in two experimental models. In the in vitro experiments, Leydig cells (LCs) were isolated from healthy mice, cultured, and exposed to increasing concentrations of Li+ (0, 10, 50, and 100 ppm). In the in vivo section of the current study, mice were treated with Li+ (0, 10, 50, and 100 ppm, in drinking water) for five consecutive weeks. Testis and sperm samples were collected and assessed. A significant sign of cytotoxicity (LDH release and MTT assay), along with disrupted testosterone biosynthesis, impaired mitochondrial indices (ATP level and mitochondrial depolarization), and increased biomarkers of oxidative stress were detected in LCs exposed to Li+. On the other hand, a significant increase in serum and testis Li+ levels were detected in drug-treated mice. Moreover, ROS formation, LPO, protein carbonylation, and increased oxidized glutathione (GSSG) were detected in both testis tissue and sperm specimens of Li+-treated mice. Several sperm anomalies were also detected in Li+-treated animals. On the other hand, sperm mitochondrial indices (mitochondrial dehydrogenases activity and ATP levels) were significantly decreased in drug-treated groups where mitochondrial depolarization was increased dose-dependently. Altogether, these data mention oxidative stress and mitochondrial impairment as pivotal mechanisms involved in Li+-induced reproductive toxicity. Therefore, based on our previous publications in this area, therapeutic options, including compounds with high antioxidant properties that target these points might find a clinical value in ameliorating Li+-induced adverse effects on the male reproductive system.

Effects of in utero and lactational exposure to the no-observed-adverse-effect level (NOAEL) dose of the neonicotinoid clothianidin on the reproductive organs of female mice.

Recently, developmental exposure to clothianidin (CLO) has been shown to cause reproductive toxicity in male mice, but the effects in female mice remain to be clarified. Pregnant C57BL/6N mice were given a no-observed-adverse-effect-level (NOAEL) dose of CLO until weaning. We then examined ovaries of 3- or 10-week-old female offspring. In the CLO-administered group, morphological changes, a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 (GPx4), and activation of genes in the steroid hormone biosynthesis pathway were observed in 3-week-old mice, and decreases of GPx4 immunoreactivity, 17OH-progesterone and corticosterone levels were observed in 10-week-old mice, along with high rates of infanticide and severe neglect, providing new evidence that developmental exposure to CLO affects juvenile and adult mice differently.

Insight into the mechanism of aspartame-induced toxicity in male reproductive system following long-term consumption in mice model.

Aspartame is one of the most common consumed artificial sweeteners utilized in many food products and beverages. It has been indicated that long-term consumption of aspartame leads to reproductive toxicity but its mechanism is not well-clear. In this study we investigated mechanism of aspartame-induced reproductive toxicity in male mice. For this purpose, 36 NMRI mature male mice received three doses of 40, 80, and 160 mg/kg body weight of aspartame, respectively per day by gavage for 90 days and also a control group was considered which received 0.5 mL of normal saline as the same route. The results revealed that long-term administration of aspartame at high doses significantly (P < .05) reduced gonadosomatic index, serum concentration of pituitary-testicular axis hormones (FSH, LH, and testosterone). It also decreased sperm parameters and total antioxidant capacity, antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase), while it caused increase in nitric oxide and malondialdehyde levels in testis tissue and sperm samples. Also, it decreased attenuated testicular histomorphometric indices (tubular differentiation index, spermiogenesis index, and repopulation index), and steroidogenic foci, while increased mRNA damages and apoptosis rate, downregulated antiapoptotic (Bcl-2) and upregulated proapoptotic (P53, BAX, and caspase-3) mediators respectively in testis. These findings indicated that consumption of aspartame for a long period results in male reproductive toxicity by decrease in serum concentration of pituitary-testis axis hormones and induction of oxidative stress and apoptosis in testis.

Manganese exposure caused reproductive toxicity of male mice involving activation of GnRH secretion in the hypothalamus by prostaglandin E2 receptors EP1 and EP2.

Exposure to manganese (Mn) can cause male reproductive damage and lead to abnormal secretion of sex hormones. Gonadotropin-releasing hormone (GnRH) plays an important role in the neuromodulation of vertebrate reproduction. Astrocytes can indirectly regulate the secretion of GnRH by binding paracrine prostaglandin E2 (PGE2) specifically to the EP1 and EP2 receptors on GnRH neurons. Prior studies assessed the abnormal secretion of GnRH caused by Mn exposure, but the specific mechanism has not been reported in detail. This study investigated the effects of Mn exposure on the reproductive system of male mice to clarify the role of PGE2 in the abnormal secretion of GnRH in the hypothalamus caused by exposure to Mn. Our data demonstrate that antagonizing the EP1 and EP2 receptors of PGE2 can restore abnormal levels of GnRH caused by Mn exposure. Mn exposure causes reduced sperm count and sperm shape deformities. These findings suggest that EP1 and EP2, the receptors of PGE2, may be the key to abnormal GnRH secretion caused by Mn exposure. Antagonizing the PGE2 receptors may reduce reproductive damage caused by Mn exposure.

Ameliorative effect of edible Halopteris scoparia against cadmium-induced reproductive toxicity in male mice: A biochemical and histopathologic study.

Cadmium (Cd) is a toxic metal affecting the reproductive system. Halopteris scoparia (brown algae) is generally consumed as a salad in the Far East countries. This study was conducted to compare and determine the possible protective effects of H.scoparia and vitamin E and C combination (VEC) against cadmium chloride (CdCl2 )-induced reproductive toxicity. A total of 36 male mice were equally divided into as control, CdCl2 (2mg/kg), CdCl2 +H.scoparia (900mg/kg), CdCl2 +VEC (200mg/kg), H.scoparia alone and VEC alone groups. Blood and testis samples were taken for biochemical, histochemical and immunohistochemical analyses. H.scoparia was also examined for antioxidant activity (by DPPH assay) and mineral/trace element content (by ICP-MS method). CdCl2 exposure caused a significant deterioration in body weight, sperm parameters (count, motility, viability and morphology) (p<.001), histopathology, immunoreactivity and testosterone levels. However, H.scoparia improved CdCl2 -induced deterioration effects more successfully than VEC-treated group. The present study suggests that edible H.scoparia can be used as a natural protective agent against Cd-induced testicular damage by possibly enhancing essential element levels or increasing antioxidant defence system.

ZnO Nanoparticles Induced Male Reproductive Toxicity Based on the Effects on the Endoplasmic Reticulum Stress Signaling Pathway.

PurposeThe aim of this study was to evaluate the adverse effects of ZnO NPs on male reproductive system and explore the possible mechanism.MethodsIn this study, the effect of oral administration of 50, 150 and 450 mg/kg zinc oxide nanoparticles (ZnO NPs) in adult male mice was studied over a 14-day period.ResultsThe results showed that the number of sperms in the epididymis and the concentration of testosterone in serum were decreased with an increased dose of ZnO NPs. Testicular histopathological lesions like detachment, atrophy and vacuolization of germ cells were observed. The results showed that increased dosage of ZnO NPs correspondingly up-regulated the IRE1α, XBP1s, BIP, and CHOP (P<0.05) which are genes related to ER stress. These observations indicated that ZnO NPs had adverse effects on the male reproductive system in a dose-dependent manner possibly through ER stress. The expression of caspase-3 was significantly increased in all the treated groups (P<0.001), which reflected the possible activation of apoptosis. Additionally, there was significant down-regulation of the gene StAR (P<0.05), a key player in testosterone synthesis. When an ER-stress inhibitor salubrinal was administered to the 450 mg/kg ZnO NPs treatment group, the damages to the seminiferous tube and vacuolization of Sertoli and Leydig cells were not observed. Furthermore, the testosterone levels in the serum were similar to the control group after the subsequent salubrinal treatment.ConclusionIt may be inferred that the ZnO NP's reproductive toxicity in male mice occurred via apoptosis and ER-stress signaling pathway.

Effect of a beta-cypermethrin and emamectin benzoate pesticide mixture on reproductive toxicity in male mice in a greenhouse environment

With the widespread use of pesticides, the resistance to pesticides of pests has gradually increased, caused mixed pesticides to become even more widely used for practical applications. To investigate the effects of mixed pesticides on reproductive health in an occupational greenhouse environment, the greenhouse environment and the characteristics of the actual application were constructed, and then the male mice were comprehensively exposed to a mixture of the beta-cypermethrin and emamectin benzoate environmental. Additionally, the effect of the beta-cypermethrin and emamectin benzoate mixture on the reproductive health of male mice was known. The results showed that with the prolongation of exposure duration, the activities of Glutathione Peroxidase (GSH-Px), Total Superoxide Dismutase (T-SOD), Lactate dehydrogenase (LDH) and Acid phosphatase (ACP) in the testes of mice gradually decreased and the activity of Malondialdehyde (MDA) gradually increased. It was also found that the apoptosis rate of murine testicular cells increased and that DNA damage occurred with prolonged exposure duration. Therefore, it can be inferred that exposure to a mixture of the pesticides beta-cypermethrin and emamectin benzoate in the greenhouse environment may have adverse effects on the reproductive health of male mice.

Reproductive toxicity assessment of Senecio scandens extract in kunming mice

This study assessed the effects of Senecio scandens Buch. Ham.extract on reproductive spectrum in female mice using a three phase reproductive assessment method. Sperm abnormality rate was used to evaluate effect on genetic reproductive toxicity. Senecio scandens extract was relatively non-toxic at low doses in all phases of reproduction in female mice. However, it showed a dose-response effect on sperm abnormality rate, and at high doses has the potential to induce genetic reproductive toxicity in male mice. It is suggested that the well-understood pharmacological benefits of S. scandens extract as an herbal medicine far outweigh the minimal reproductive toxicity effects observed in this study.

The flavonoid chrysin protects against zearalenone induced reproductive toxicity in male mice

The mycotoxin zearalenone (ZEA) has strong estrogenic effects and elicits reproductive toxicity. Chrysin is a natural flavonoid found in many plant and has a broad range of pharmacological activities, including anticancer, antioxidant and anti-inflammatory. The present study aimed to investigate the potential protective effects of chrysin against ZEA toxicity. Mice received chrysin (5 or 20 mg/kg; i.g.) for ten days, and then received a single injection of ZEA (40 mg/kg). Two days thereafter, blood and testes were collected. ZEA decreased number and motility of sperm, plasma testosterone levels, enzymatic (glutathione peroxidase, glutathione reductase, glutathione-S-transferase) and non-enzimatic defenses (reduced glutathione). Moreover, ZEA increased 4-hydroxynonenal and 8-hydroxy-2′-deoxyguanosine levels, myeloperoxidase activity and levels of proinflammatory cytokines (interleukins-1β and 6, tumor necrosis factor alpha). ZEA also decreased levels of anti-inflammatory cytokine interleukin-10 and increased activity of caspases 3 and 9. Chrysin treatment increased the number and motility of sperm, testosterone levels, restored antioxidant defenses and reduced the inflammation and apoptosis process. In summary, chrysin attenuated the toxic effects caused by ZEA in blood and testes of mice, suggesting a potential preventive treatment against the deleterious effects of ZEA.

From the Cover: Roles of mmu_piR_003399 in Microcystin-Leucine Arginine-Induced Reproductive Toxicity in the Spermatogonial Cells and Testis.

Our previous work has demonstrated that microcystin-leucine arginine (MC-LR) is a potent toxin for the reproductive system of male mice and it exerts cytotoxicity in spermatogonial cells, resulting in the constitutional and functional changes of the mouse testis. The present study was designed to investigate the functions of P-element-induced wimpy (piwi)-interacting RNAs (piRNAs) in MC-LR-induced reproductive toxicity in male mice. We observed an increase in the mmu_piR_003399 level in spermatogonial cells and mouse testicular tissues following treatment with MC-LR. Moreover, our data confirmed that cyclin-dependent kinase 6 (CDK6) was the target gene of mmu_piR_003399. Increases in the concentration of mmu_piR_003399 were correlated with the reduced expression of CDK6 both in vitro and in vivo. mmu_piR_003399 induced cell cycle arrest at the G1-phase, down-regulated sperm counts and sperm motility, and compromised sperm morphology. On the contrary, suppressing the expression of mmu_piR_003399 could substantially attenuate MC-LR-induced pathology in mice including cell cycle arrest, reduced mature sperm counts, sperm viability loss and abnormal sperm morphology. Furthermore, our data supported that mmu_piR_003399 existed in mouse serum and plasma, and its level was increased in MC-LR-treated mice. In conclusion, we demonstrate that mmu_piR_003399 plays a crucial role in regulating MC-LR-induced reproductive toxicity.

Proanthocyanidin protects against acute zearalenone-induced testicular oxidative damage in male mice.

Zearalenone (ZEN) exerts a major effect on human and animal health and has led to serious worldwide economic problems. In this study, we investigated whether proanthocyanidin (PC) can prevent ZEN-induced testicular oxidative damage in male mice and explored the underlying mechanism. Kunming mice were injected with ZEN (40mgkg-1) on the 11th day after intragastric administration of PC (75 or 150mg/kg) for 10days; the sperm quality of mice was then analysed statistically. Additionally, testicular morphology parameters related to oxidative damage, apoptosis and the expression of endoplasmic reticulum (ER) stress-related genes (GRP78, CHOP and XBP-1) were all measured. Results showed that ZEN exposure significantly reduced the sperm density, improved the sperm aberration rate, increased the MDA level and reduced SOD and GSH-Px activities. Meanwhile, ZEN was attributed to the downregulation of the expressions of the gene and protein of Bcl-2 and upregulation of the expressions of the gene and protein of Bax and caspase-3. ZEN exposure also upregulated the mRNA expression of GRP78, CHOP and XBP-1; however, PC pre-treatment reduced ZEN-induced oxidative damage and tended to maintain normal testicular morphology. Furthermore, PC pre-treatment substantially downregulated the expressions of the GRP78, CHOP and XBP-1 and upregulated the expression of the Bcl-2 gene. In conclusion, PC, due to its anti-oxidative ability, could ameliorate ZEN-induced testicular reproductive toxicity in male mice by decreasing ER stress and testicular cell apoptosis.