Testicular Index Research Articles

Myricetin reduces the reproductive toxicity of cyclophosphamide in male mice

To explore the effect of myrica flavone on male reproductive toxicity induced by cyclophosphamide and its mechanism. Thirty 6-week-old male ICR mice were randomly divided into 5 groups: blank control group, cyclophosphamide reproductive injury model group, myricetin low-medium high-dose intervention group. Except the blank control group, the other groups were intraperitoneally injected with cyclophosphamide 50 mg/kg daily for 7 consecutive days. The myricetin group received intragastric administration of 100, 200, and 400 mg/kg myricetin daily for 30 consecutive days since the second day of modeling. The blank control group and the model control group were given an equal volume of a 0. 25% sodium carboxymethyl cellulose solution. The body weight was measured every 3 days, and the day after the last administration, the mice were sacrificed by cervical dislocation, and the epididymis and testes were quickly taken. Testicular weighing, testicular index calculation, epididymis to obtain sperm, sperm analyzer to analyze sperm density and vitality. The expression of Bax and Bcl-2 in testicular tissues was detected by immunoblotting, and the mitochondrial membrane potential of sperm was detected by flow cytometry. After 9 days of modeling, the weight of mice in the model group was lower than that of the blank control group, which was statistically different(P<0. 05). There was no difference between the myricetin treatment group and the model group. The testis index of the model group was(3. 93±0. 91)mg/g, which was significantly lower than that of the blank control group(6. 93±0. 98)mg/g, and the difference was statistically significant(P<0. 05). After treatment with bayberry flavonoids, the testis index increased, in the 100 and 200 groups and 400 mg/kg testis index were(3. 94±1. 21) mg/g, (4. 33±0. 88) mg/g, and(4. 80±0. 43) mg/g, respectively. Compared with model control group, The difference was statistically significant(P<0. 05 and P<0. 01). Compared with the control group, the sperm density, sperm rate of forward movement, sperm rate of non-forward movement, and decreased sperm rate of non-moving sperm increased in the model group. After treatment with bayberry flavonoids, compared with the model group, the sperm density, sperm rate of forward motion, and sperm rate of non-forward motion increased, and the immobility sperm rate decreased. The 200 and 400 mg/kg groups had statistical significance(P<0. 05 or P<0. 01); the normal rate of sperm mitochondrial membrane potential in the model group was(54. 70±5. 45)%, and the normal mitochondrial membrane potential rate after treatment with myricetin of 100, 200 and 400 mg/kg(59. 10±9. 97)%, (62. 10±6. 07)% and(77. 10±8. 87)%, of which the 400 mg/kg group was statistically significant(P<0. 05); the ratio of Bax/Bcl-2 in the model group was 5. 92±1. 45, and the ratio of Bax/Bcl-2 decreased after treatment with myricetin of 100, 200 and 400 mg/kg, which were 2. 52±0. 51, 1. 71±0. 52 and 1. 07±0. 29. There were statistical differences(P<0. 05 or P<0. 01). Myrica flavone can protect sperm mitochondrial membrane potential, inhibit testicular cell apoptosis, and protect the male mice from reproductive toxicity induced by cyclophosphamide.

Assessment of the function of gonad-specific PmAgo4 in viral replication and spermatogenesis in Penaeus monodon

Argonaute family is phylogenetically subdivided into Ago and Piwi subfamilies that operate either transcriptional or post-transcriptional regulation in association with particular types of small RNAs. Among the four members of Ago subfamily (PmAgo1-4) found in black tiger shrimp Penaeus monodon, PmAgo4 exhibits gonad-restricted expression and takes part in transposon repression as the Piwi subfamily. While PmAgo1-3 participate in RNA interference (RNAi)-based mechanism, the role of PmAgo4 in RNAi is still mysterious, and was therefore investigated in this study. The results showed that knockdown of PmAgo4 in shrimp testis did not have a significant effect on the potency of PmRab7 silencing by dsPmRab7. In addition, replication of YHV as well as YHV-induced cumulative mortality in PmAgo4-knockdown shrimp are comparable to the control shrimp, suggesting the irrelevant association of PmAgo4 with RNAi-mediated gene silencing and antiviral immunity. Since PmAgo4 did not function in common with the Ago subfamily, its potential function in gametogenesis of male shrimp was further investigated. The reduction of PmAgo4 transcript levels in male shrimp revealed significant defect in testicular maturity as measured by Testicular Index (TI). Moreover, the numbers of mature sperm in spermatophore of PmAgo4-knockdown shrimp were significantly decreased comparing with the control shrimp. Our studies thus suggest a distinctive role of PmAgo4 that is not consistent with a dsRNA-mediate gene regulation and virus replication, but has a key function in controlling spermatogenesis in P. monodon.

Rubus apetalus (Rosaceae) improves spermatozoa characteristics, antioxidant enzymes and fertility potential in unilateral cryptorchid rats

BackgroundCryptorchidism (CPT) is an important cause of male infertility. Rubus apetalus is a medicinal plant with a powerful antioxidant potential. We investigated the effects of aqueous and methanolic extracts of R. apetalus on spermatozoa parameters, antioxidant enzymes and fertility potential of rats with experimental unilateral CPT.MethodNormal (n = 15), sham-operated (n = 15) and cryptorchid rats (n = 80; distributed into 16 groups of 5 rats/group) were treated for 2, 4 or 8 weeks with either distilled water (10 ml/kg/day), vitamin E (75 mg/kg/day), aqueous or methanolic extract of R. apetalus (12 and 60 mg/kg). Sex organ weights, spermatozoa parameters, testicular proteins, sex hormones, fertility potential, morphometric characteristics of testis and oxidative stress markers were measured.ResultsCPT significantly (p < 0.05–0.001) decreased testicular and epididymal weights, spermatozoa density, spermatozoa motility, spermatozoa normality, testicular proteins, LH, FSH and testosterone concentrations. In cryptorchid rats, peri-vascular fibrosis significantly increased (p < 0.001), while diameter of the seminiferous tube, germ cell thickness, gestation index and fertility index decreased when compared to control. Additionally, CPT induced oxidative stress by increasing lipid peroxidation and by reducing superoxide dismutase and catalase activities. These alterations were corrected by R. apetalus. For instance, a significantly increase (p < 0.05–0.001) in spermatozoa motility, normality, viability and density after 2, 4 and 8 weeks of treatment was noticed. R. apetalus also increased (p < 0.05–0.001) testicular proteins, gestation index (90–100%) and fertility index (90–100%), compared to the untreated cryptorchid rats.ConclusionR. apetalus boosts fertility potential in cryptorchid rats and could be considered as a promising alternative agent for the management of infertility associated with CPT.

Effects Of Exercise On The Expression Of Kiss-1/GPR54 In Testis Of High-fat Diet Rats In Growth Phase

PURPOSE: To explore the effect of exercise on localization and expression of KISS-1 and GPR54 in the growth period (PND 21st to 56th day) of high-fat diet rat testes. METHODS: 21D old weanling rats were randomly divided into group HC (n=32) and group HE (n=32). HE group took 5-weeks trained (60-70% vVO2max, 1h/day, 5days/week). Groups HC and HE were fed with high-fat feed, which was purchased from Beijing Huafukang Biotechnology Co. Ltd. (LOT No: D12451).The rats of two groups were killed on the 21st D, 35th D, 43rd D, and 56th D old. The localization, mRNA expression and protein expression of KISS-1/GPR54 in the testis of each group were tested. RESULTS: The high-fat diet intervention resulted in a decrease in testicular index from pre-puberty(p<0.05); testicular tissue testosterone decreased significantly from prepuberty(p<0.01), and endocrine function was impaired; rat testicular tissue structure was loose, and the number of mature sperm decreased. The high-fat diet intervention had no significant effect on the localization of KISS-1/GPR54 system in rat testis, but affected the expression of KISS-1/GPR54 in rat testis and down-regulated the expression of KISS-1 protein(p<0.01). Exercise intervention can improve testicular development in rats with high-fat diet, especially to improve sex hormone disorders caused by high-fat diet. Under exercise intervention, the testicular tissue testosterone decline caused by high-fat diet was improved(p<0.05). Comparing with HC group, the expression of GPR54 protein increased significantly(p<0.01),but the KISS-1 protein expression had no obvious change(p>0.05). CONCLUSION: High-fat diet can inhibit the testicular development of male rats in the growth phase, and can also down-regulate the protein and gene expression levels of KISS-1/GPR54 system in testis tissue, and change the expression of KISS-1/GPR54 system, and its role needs further study. 60~70% VO2max moderate-intensity aerobic exercise change the inhibitory effect of high-fat diet on testicular development in male rats, and up-regulate KISS-1/GPR54 in prepubertal stage and whether KISS-1/GPR54 in testicular tissue participates in its regulation remains to be further study.

Photoperiod affects gamete production, and protein and lipid metabolism in male narrow-clawed Crayfish Pontastacus leptodactylus (Eschscholtz, 1823)

Understanding effects of photoperiod on the reproductive efficiency and physiological responses of Pontastacus leptodactylus is important for successful management of the crayfish hatcheries. Spermatozoal number, gonado-somatic index (GSI), hepatosomatic index (HSI), testicular index (TI), vas deferens index (VDI), and total protein, amino and fatty acid profiles of the hepatopancreas were evaluated in male narrow-clawed crayfish P. leptodactylus. Animals (30–70 g) were maintained utilizing five different photoperiodic regimens (3 L:21D, 6L:18D, 24 L:0D, 0L:24D, and natural photoperiod (9 L:15D)) for 5 weeks. Different photoperiods affected spermatozoal number, GSI, and total protein, amino acid and fatty acid profiles of the hepatopancreas (P < 0.05). The greatest spermatozoal number and GSI were recorded in darkness (P < 0.05). There was the least spermatozoal number (3.5 × 106 ± 0.5) and GSI (2 ± 0.1%) when imposing the 9 L and 24 L photoperiodic regimens, respectively. In addition, maintenance in constant light as compared with other lighting regimens resulted in a greater total protein (43.68 ± 4.83 mg/g), phenylalanine (7.23 ± 0.35%), and total single unsaturated fatty acids (ΣMUFA) (48.07 ± 2.30%), but lesser total polyunsaturated fatty acids (ΣPUFA) (23.36 ± 1.07%), serine (1.53 ± 0.02%), Σn-6 (11.24 ± 1.18%), 20:4 (n-6) (2.7 ± 0.14%), and 20:2 (0.67 ± 0.17%) contents in the hepatopancreas (P < 0.05). The results of the present study indicate total darkness increases the spermatozoal production and imposing longer dark periods improved the reproductive efficiency and physical conditions in male P. leptodactylus.

Effect of serotonin injection on the reproductive parameters and haemolymph methyl farnesoate level in the narrow‐clawed crayfishPontastacus leptodactylus(Eschscholtz, 1823)

The effects of serotonin injection (1, 5 and 10 µg/g body weight) on spermatozoal production, vas deferens index (VDI), hepatosomatic index (HSI), testicular index (TI), gonado-somatic index (GSI), ovarian egg number, total protein, methyl farnesoate (MF) levels of haemolymph, amino acid and fatty acid profiles of hepatopancreas were evaluated in Pontastacus leptodactylus. In male P. leptodactylus, the highest spermatozoal number (9.3 × 106 ± 1.2) and MF level (0.66 ± 0.06 ng−1 ml) were observed after injection of 10 µg/g of serotonin (p < .05). Serotonin injection, however, did not significantly affect the reproductive parameters in female P. leptodactylus (p > .05). It was also found that serotonin injection caused an increase in the level of MF in male P. leptodactylus but a reduction in female P. leptodactylus (p < .05). Therefore, a sex-dependent response to serotonin could exist for MF synthesis in P. leptodactylus. Serotonin injection did not affect the total protein, amino acid and fatty acid profiles in the hepatopancreas of male P. leptodactylus (p > .05). It can be concluded that serotonin injection affects the reproductive parameters such as spermatozoal production, VDI, TI, GSI, HSI and haemolymph MF levels in P. leptodactylus.

Cuscuta chinensis flavonoids alleviate bisphenol A-induced apoptosis of testicular cells in male mice offspring.

Bisphenol A (BPA) is a widespread environmental endocrine disruptor that has multiple effects on reproductive organ development. To investigate the effect of Cuscuta chinensis flavonoids (CCFs) on testicular apoptosis induced by BPA in male mice offspring, pregnant mice were administered intragastrically with BPA and CCF at gestation day (GD) 0.5-17.5. The testes of male offspring (F1 males) were collected at post-natal day (PND) 21 and PND 56 for the detection of related indicators. The results showed that compared with the BPA group, the testicular index in CCF groups was significantly increased at PND 21 (p<.01). For the mice of different concentrations of CCF groups, the expression levels of bax, caspase-9 and caspase-7 proteins were significantly decreased at PND 21 and PND 56, while the expression level of bcl-2 protein was significantly increased, and testicular apoptotic cells were also decreased significantly (p<.01 or p<.05). Forty mg/kg CCF has no significant difference compared with the control group. The results indicated that CCF could protect the testis development of F1 male mice by alleviating the apoptosis of testicular cells induced by BPA.

Effects of Carob Fruit Extract on Spermatogenesis, Antioxidant Status, and Apoptosis in Adult Male Mice

Background: Certain plants stimulate spermatogenesis and increase fertility; in contrast, some plants arrest the spermatogenesis cycle. Ceratonia siliqua is an herb plant with a strong antioxidant property. The aim of this study was to evaluate the effects of carob fruit extract on spermatogenesis, testicular apoptosis, and oxidative stress in adult male mice. Methods: Forty adult male mice were randomly divided into five groups: control, sham, and carob 1–3. The sham group was injected with normal saline and the carob 1–3 groups were injected with 200, 400, and 800 mg/kg of the carob fruit extract intraperitoneally for 14 days, respectively. At the end of the injection period, spermatogenesis, testicular apoptosis, and oxidative stress were examined. Results: The sperm parameters increased in the mice that received 200 mg/kg of carob compared to the sham group (p &lt;0.05). There was a significant increase in the weight index of the epididymis in the carob 3 group in comparison to the sham group (p = 0.01). The number of positive tunnel cells was not statistically significant between different groups (p&gt;0.05). The level of malondialdehyde decreased in the carob 1 and carob 3 groups, but this reduction was not statistically significant (p &gt; 0.05). In addition, the statistical analysis showed a significant difference in the mean superoxide dismutase levels in the carob 2 and carob 3 groups in comparison to the sham group (p ≤ 0.001). The statistical analysis showed a significant increase in the mean level of the catalase enzyme in the carob 1 group in comparison to the sham (p = 0.02), and carob 2 groups (p = 0.008). Conclusion: The administration of 200 mg of the carob fruit extract for 14 days increased the testicular index as well as sperm parameters and decreased the level of oxidative stress in the testicular tissue of adult mice.

Maternal exposure to bisphenol A during pregnancy interferes testis development of F1 male mice.

This study was conducted to investigate the effects of maternal exposure to bisphenol A (BPA) on testis development of F1 male mice. The BPA exposure model of pregnant mice was prepared by intragastric administration of BPA at the doses of 0, 2.5, 5, 10, 20, and 40mg/kg/day at gestation day (GD) 0.5-17.5. The testis index of the offspring mice was calculated at postnatal day (PND) 21 and PND 56. The results showed that maternal exposure to 20mg/kg BPA during pregnancy significantly increased the testicular index of F1 males at PND 21, and 40mg/kg BPA significantly decreased the testicular index of F1 males at PND 56 (P < 0.01). BPA significantly reduced serum testosterone (T) and estradiol (E2) levels, and improved testicular ERα and ERβ levels in F1 males at both PND 21 and PND 56. BPA exposure also upregulated transcription of testicular Dnmt1 and inhibited the transcription of testicular Dnmt3A and Dnmt3B in F1 mice at PND 21. BPA reduced the transcriptional level of testicular DNA methyltransferase (Dnmt), increased the expression of testicular caspase-7, caspase-9, and bax, and decreased the expression of bcl-2 in F1 mice at PND 56. Consistent with that, BPA improved the apoptosis rate in the testis at PND 56 (P < 0.01 or P < 0.05). Our study indicates that BPA disrupts the secretion of testosterone, estradiol, and estrogen receptors by interfering with the transcription of testicular DNA methyltransferase (Dnmt) in offspring males, which damages testicular tissues and affects the potential reproductive function.

The annual cycle of spermatozoa content in the vas deferens and some reproductive parameters in the narrow-clawed crayfish Pontastacus leptodactylus (Eschscholtz, 1823)

Understanding the annual dynamic of gametes in crayfish is useful for successful management of crayfish population. The annual variations in spermatozoa content of vas deferens, gonado-somatic index (GSI), hepatosomatic index (HSI), vas deferens index (VDI), and testicular index (TI) were investigated in male Pontastacus leptodactylus. The spermatozoa content, GSI, HSI, VDI, and TI of P. leptodactylus significantly (P < 0.05) changed during different seasons of the year. For example, significantly higher spermatozoa number, GSI, VDI, and TI were recorded in January (P < 0.05). On the other hand, the HSI reached the peak in August (P < 0.05). The spermatophore degeneration started in April. Although the vas deferens lacks spermatophore structure during the summer, the presence of spermatozoa was observed throughout the year. This study represents a step forward in understanding the seasonal spermatozoa content and the reproductive status of male P. leptodactylus.

Effect of administration of neurotransmitters: Serotonin and dopamine on testicular maturation in the freshwater crab Travancoriana schirnerae Bott, 1969 (Crustacea: Gecarcinucidae)

The current investigation aimed at the effect of administration of the neurotransmitters, serotonin and dopamine, on testicular maturation in the freshwater crab Travancoriana schirnerae Bott, 1969 (Crustacea: Gecarcinucidae). Adult males in active, inactive and revival phases of spermatogenesis were injected with serotonin and dopamine in multiple doses and their dissected testes were submitted to histomorphological examinations. Though serotonin treatment during active phase showed increased values for testicular index, acinar diameter and proportion of mature spermatozoa, only the testicular index values differed statistically from that of controls. Serotonin administration during inactive phase caused an increase in testicular index and acinar diameter, reduction in intra and interacinar spaces and pycnosis of germ cells and detection of division stages. A notable increase in the testicular index, acinar diameter and fresh batches of spermatozoa was perceptible in crabs injected with serotonin during revival phase. In contrast, dopamine injection showed a decline in testicular activity irrespective of the phase of spermatogenesis, as evinced from low testicular index, acinar diameter, presence of small, irregularly shaped acini with indistinct acinar boundaries, reduction in the number of mature spermatozoa and absence of division stages. To conclude, the stimulatory neurotransmittant, serotonin can be used to induce testicular maturation in species of aquaculture potential.

Short-term toxicity of dibutyl phthalate to mice intestinal tissue.

The objective of this study was to investigate changes in intestinal histopathology and expression of heat-shock proteins (HSPs) in the small intestinal tissue of mouse after acute exposure to dibutyl phthalate (DBP). Forty-eight 60-day-old Institute of Cancer Research (ICR) mice were administered DBP by gavage once a day for 10 days. The mice were divided into three groups of 16 mice each: the high-dose group was administered 500 mg/kg body weight (BW) DBP; the low-dose group was administered 50 mg/kg BW; and the control group was not administered DBP. Significant increases in the uterine index, ovary index, and testicular index were observed in the DBP-exposed groups compared to those in the control group. Villus height and V/ C ratio significantly increased ( p < 0.05) in the duodenum and decreased ( p < 0.05) in the jejunum after the administration of DBP. The goblet cell number decreased in both the duodenum and the jejunum of mice exposed to DBP ( p < 0.05) compared to the number in the control group mice. Damage to the structure of the small intestine was accompanied by a marked increase in HSP27 expression and a decrease in the expression of HSP70 and HSP90 in both high-dose and low-dose groups. These results indicate that elevated HSP27 levels in the duodenum and jejunum may be important markers for acute DBP exposure and that HSP27 may act as a protective protein involved in intestinal mucosa repair.

Biochemical and immunocytochemical studies of the testicular alteration caused by Amitriptyline in adult male rat

Amitriptyline is one of the first reference tricyclic antidepressants (TCAs) with sedative and analgesic properties. This study aimed to investigate the testicular indices that influenced by amitriptyline treatment and examined it by biochemical, histological and Immunohistochemical methods. A total of 20 male albino rats were equally divided into two groups: the first was control and the second was amitriptyline intoxicated group. The results of this study showed that amitriptyline consumption significant decrease hormones parameters, increase sperms abnormalities, decrease sperm numbers, induced testicular tissue damage and P53 alterations.

Effects of the Endocrine-Disrupting Chemicals, Vinclozolin and Polychlorinated Biphenyls, on Physiological and Sociosexual Phenotypes in F2 Generation Sprague-Dawley Rats.

Background:Exposure to endocrine-disrupting chemicals (EDCs) during gestation influences development of the F1 generation offspring and can result in disease and dysfunction in adulthood. Limited evidence suggests consequences on the F2 generation, exposed as germ cells within the F1 fetus. These F2s provide a unique window into the programming effects of EDCs.Objective:This study assessed intergenerational effects of EDC exposure on adult physiology and behavior in Sprague-Dawley rats.Methods:Pregnant rats were exposed to either a polychlorinated biphenyl (PCB) mixture, Aroclor 1,221 (A1221), the fungicide vinclozolin (VIN), or the vehicle (VEH) (6% dimethylsulfoxide in sesame oil) alone. A1221 is weakly estrogenic, while VIN is antiandrogenic, enabling us to compare different classes of EDCs. The F1 male and female offspring were bred to generate the paternal- and maternal-lineage F2 generation. This F2 generation was assessed for physiological outcomes, ultrasonic vocalizations (USVs), and sexual behavior in adulthood.Results:Each EDC caused phenotypic effects in a sex- and lineage-dependent manner. The most robustly affected group was the paternal-lineage males. F2 VIN paternal male descendants had increased body weight throughout the lifespan, lower concentrations of circulating estradiol, and lower adrenal and testicular indices. Both VIN and A1221 paternal-lineage males also exhibited the greatest number of changes in the characteristics of USVs in response to an opposite-sex animal and changes in sexual behaviors in a mating test.Conclusion:Exposure of rats to EDCs at the germ cell stage led to differences in the physiological and behavioral phenotype later in life, especially in males. This finding has implications for multigenerational physiological and reproductive health in wildlife and humans. https://doi.org/10.1289/EHP3550

Effects of Administration of Monosodium Glutamate and Methylphenidate on the Histologic Structure of Testicular Tissue and Microscopic Indices of Spermatogenesis in Adolescent Rats

Introduction: Methylphenidate (MPH) is one of the common medications used for maintaining alertness and improving attention. Monosodium glutamate (MSG) is a food additive, which acts as an enhancer of palatability. The aim of this study was to evaluate the various effects of these compounds on the reproductive system during adolescence. Materials and Methods: Methylphenidate (5 and 10 mg/kg) and monosodium glutamate (6 and 60 mg/kg) were administrated to adolescent rats. After 60 days, the pituitary-testicular axis hormones were assayed and testicular histomorphometric studies were performed. Findings: The coadministration of MPH (5 mg/kg) and MSG (60 mg/kg) led to elevation in serum FSH levels (P less-than 0.05). The highest blood level of luteinizing hormone was observed following high doses of MPH and MSG separately or in combination form. The effect of MPH and MSG on serum testosterone level was observed dose dependently and contradictorily (P greater-than 0.05). The administration of MSG led to a reduction in the population of Sertoli cells, spermatogonia, and spermatocyte. Coadministration of MPH and MSG also reduced this population. Morphometric studies revealed decrement of tubular diameter and germinal epithelium height (P less-than 0.05), especially with low doses of MPH and high doses of MSG. The changes in spermatogenic indices were similar to morphometric results. Tubular atrophy, interstitial edema, and depopulation of spermatogenic cells were observed in MPH and MSG treated groups. Coadministration of these compounds increased spermatids with pyknotic nucleus. Conclusion: It has been concluded that the coadministration of MPH and MSG through the induction of some hormonal and structural alterations could induce some changes in the normal structure and function of the reproductive system.

Hesperetin, a citrus flavonoid, attenuates testicular damage in diabetic rats via inhibition of oxidative stress, inflammation, and apoptosis

AimThis study was designed to assess the beneficial effect of hesperetin on diabetes-associated testicular injury in the rat. Main methodsOral treatment with hesperetin started 10 days after diabetes induction by streptozotocin (60 mg/kg, i.p.) for 46 days. Testicular damage was evaluated by histological evaluation of seminiferous tubules in addition to assessment of epididymal sperm count, motility, and viability. In addition, testicular biomarkers of apoptosis, inflammation, and oxidative stress were also determined. Key findingsHesperetin treatment of diabetic group prevented body weight loss and reduced serum glucose in addition to improvement of serum testosterone. Additionally, hesperetin-treated diabetic group had lower levels of malondialdehyde (MDA), reactive oxygen species (ROS), protein carbonyl, DNA fragmentation, and caspase 3 activity as specific biomarkers of oxidative stress and/or apoptosis. Furthermore, hesperetin augmented testicular antioxidant system as shown by higher levels of glutathione (GSH), mitochondrial membrane potential (MMP), and ferric reducing antioxidant power (FRAP) in addition to improvement of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx). Moreover, hesperetin administration to diabetic rats attenuated testicular indices of inflammation consisting of tumor necrosis factor α (TNFα) and interleukin 17 (IL-17) and prevented damage of seminiferous tubules as revealed by higher levels of sperm count, motility, and viability in diabetic rats. SignificanceCollectively, hesperetin could alleviate testicular damage in DM, at least through inhibition of apoptosis, oxidative stress, and inflammation in addition to its up-regulation of endogenous enzymatic and non-enzymatic antioxidants.

Influence of season and climatic variables on testicular cytology, semen quality and melatonin concentrations in crossbred bucks reared under subtropical climate.

Seasonality in reproduction and effects of climatic variables on testicular cytology and semen quality in bucks reared under subtropical climatic conditions were not well understood. In the present study, using testicular cytology, semen evaluation and melatonin concentrations assessed over a period of 1year, we report that bucks reared under subtropical climatic conditions did not show seasonality in reproduction. Climatic variables including temperature, relative humidity, temperature-humidity index (THI), sunshine hours and day length were recorded daily during the whole period of experimentation (one complete year). Ejaculates were collected from crossbred (Alpine X Beetal) males (n = 6) biweekly using artificial vagina, and semen quality (volume, mass activity, sperm concentration, motility, viability, membrane integrity and protamine deficiency) was assessed. To understand the seasonal influence at testicular level, using fine needle aspiration biopsy method, testicular cells were aspirated and different types of cells and testicular cytology indices were quantified. Blood was collected biweekly for estimation of melatonin concentrations. Mass activity was higher (P < 0.05) during rainy season while individual sperm motility and sperm concentration were higher (P < 0.05) during rainy and autumn seasons as compared to other seasons. Sperm functional parameters did not show any differences during different seasons. Sertoli cell count, spermatogenic cell count and testicular indices did not differ among the seasons. Melatonin concentrations also did not differ significantly among the four seasons studied. Among the climatic parameters, THI had significant (P < 0.05) influence on sperm quality. The proportion of Sertoli cell in the testicular cytology had a significant and positive relationship with RH, THI and day length. It was concluded that seasonal variations are less evident in terms of spermatogenesis and semen quality in Alpine X Beetal crossbred bucks reared under subtropical climatic conditions.

Effects of perfluorooctanoic acid exposure during pregnancy on the reproduction and development of male offspring mice.

This study was conducted to explore the effects of maternal exposure to perfluorooctanoic acid (PFOA) on reproduction and development of male offspring mice. Pregnant mice were given 1, 2.5 or 5mg/kg BW PFOA daily by gavage during gestation. The results showed that the survival number of offspring mice at weaning was significantly decreased. There were no differences in the testicular index of offspring mice between PFOA exposure groups and non-PFOA group. Maternal exposure to PFOA reduced the level of testosterone in the male offspring mice on PND 21 (p<0.01) but increased in 1mg/kg group and decreased in 2.5 and 5mg/kg groups on PND 70 (p<0.01). There were different degrees of damage to testis in a dose-dependent manner, and the number of Leydig cells markedly decreased (p<0.01) in 2.5 and 5mg/kg PFOA groups on PND 21 and PND 70. The expression of Dlk1-Dio3 imprinted gene cluster showed a decreasing trend, where Glt2, Rian and Dio3 gene expressions were significantly reduced (p<0.05) on PND 21. Therefore, PFOA exposure during pregnancy reduces the number of survival offspring mice, damages testis, disrupts reproductive hormones and reduces the mRNA expressions of the Dlk1-Dio3 imprinted cluster in testis.

Comparative Profiling of MicroRNAs Reveals the Underlying Toxicological Mechanism in Mice Testis Following Carbon Ion Radiation.

This study investigated the toxicity of heavy ion radiation to mice testis by microRNA (miRNA) sequencing and bioinformatics analyses. Testicular indices and histology were measured following enterocoelia irradiation with a 2 Gy carbon ion beam, with the testes exhibiting the most serious injuries at 4 weeks after carbon ion radiation (CIR) exposure. Illumina sequencing technology was used to sequence small RNA libraries of the control and irradiated groups at 4 weeks after CIR. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses implicated differential miRNAs in the regulation of target genes involved in metabolism, development, and reproduction. Here, 8 miRNAs, including miR-34c-5p, miR-138, and 6 let-7 miRNA family members previously reported in testis after radiation, were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) to validate miRNA sequencing data. The differentially expressed miRNAs described here provided a novel perspective for the role of miRNAs in testis toxicity following CIR.

Ginsenoside Rg1 ameliorates testicularsenescencechanges in D‑gal‑induced aging mice via anti‑inflammatory and antioxidative mechanisms.

With the growing population, aging, extended lifespans and anti-aging have become popular areas of research in the life and social sciences. With increasing age, the structure and function of the testes, the spermatogenetic and androgen-producing organ in the male reproductive system, gradually declines. Ginsenoside Rg1 is an extract of Panax ginseng in traditional Chinese medicine. The extract facilitates anti-aging through its anti-inflammatory and antioxidant properties. However, it has not been reported whether ginsenoside Rg1 delays testicular aging. The present study established D-galactose (D-gal)-induced aging mouse models to examine the protective effects of ginsenoside Rg1 on the structure and function of the testes, and the underlying mechanism. A total of 60 healthy specific pathogen-free male C57BL/6 mice were randomly divided into four groups: Control group; Rg1 group; D-gal + Rg1 group; and D-gal group. The tissues of the mice were used for further experiments. The present study further investigated the effects of Rg1 on the volume of serum testosterone, the testicular index, testicular microscopic structures, the senescence of spermatogenetic cells, the apoptosis of spermatogenetic cells, the activity of the antioxidant enzymes, the levels of inflammatory cytokines, and the levels of S-phase kinase-associated protein (p19), cyclin-dependent kinase inhibitor 1 (p21) and cellular tumor antigen p53 (p53) in D-gal-induced aging mice. In general, compared with the D-gal group, the treatment of Rg1 increased the testis index, serum testosterone level and the active content of superoxide dismutase and the total antioxidant capacity. The percentage of senescence-associated β-galactosidase-positive cells, the level of apoptosis and the volume of methane dicarboxylic aldehyde, tumor necrosis factor-α, interleukin (IL)-1β and IL-6 in testicular tissues were significantly decreased, and the expression of p19, p53 and p21 was downregulated due to the treatment with Rg1. The results of the present study demonstrated that ginsenoside Rg1 was able to protect the testes against D-gal-induced aging in mice. In addition, the protective effect of Rg1 may be achieved via antioxidation and downregulation of the p19/p53/p21 signaling pathway.