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1.
Int J Mol Sci ; 20(18)2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31510090

RESUMO

The negative association between psychological stress and male fertility has been known for many years. This study was aimed at (i) identifying spermatogenesis impairment induced by psychological stress in rats and (ii) exploring the role of glucocorticoid receptor (GR) signaling in these adverse effects (if they exist). Male Sprague Dawley rats were exposed to a six-week period of unpredictable chronic mild stress (uCMS) along with cotreatment of GR antagonist RU486 (1 mg/kg/day). Testicular damage was assessed by testicular pathological evaluation, epididymal sperm concentration, serum testosterone levels, testicular apoptotic cell measurements, and cell cycle progression analyses. Rats in the uCMS group had decreased levels of serum testosterone and decreased epididymal sperm concentration. The uCMS-treated rats also had decreased numbers of spermatids and increased levels of apoptotic seminiferous tubules; additionally, cell cycle progression of spermatogonia was arrested at the G0/G1 phase. Furthermore, uCMS exposure caused an increase in serum corticosterone level and activated GR signaling in the testes including upregulated GR expression. RU486 treatment suppressed GR signaling and alleviated the damaging effects of stress, resulting in an increased epididymal sperm concentration. Overall, this work demonstrated for the first time that the activation of GR signaling mediates stress-induced spermatogenesis impairment and that this outcome is related to cell apoptosis and cell cycle arrest in germ cells.


Assuntos
Epididimo/metabolismo , Receptores de Glucocorticoides/metabolismo , Espermatogênese/fisiologia , Espermatozoides/metabolismo , Estresse Psicológico/fisiopatologia , Testículo/metabolismo , Animais , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Epididimo/citologia , Epididimo/efeitos dos fármacos , Masculino , Mifepristona/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Glucocorticoides/antagonistas & inibidores , Túbulos Seminíferos/citologia , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/metabolismo , Contagem de Espermatozoides , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/citologia , Testículo/efeitos dos fármacos , Testosterona/sangue
2.
J Pharm Pharmacol ; 71(10): 1565-1575, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31385305

RESUMO

OBJECTIVES: This study was designed to assess the role of caffeine on fertility parameters in testicular and epididymal tissues of scopolamine-induced model of amnesia in rats. METHODS: Adult male rats were treated with scopolamine with or without caffeine. The modulatory effects of caffeine or scopolamine on fertility parameters were assessed in rats' testicular and epididymal homogenates. KEY FINDINGS: Scopolamine-induced sperm abnormalities, reduced steroidogenic enzyme 3ß-Hydroxysteroid dehydrogenase (3ß-HSD) and 17ß-Hydroxysteroid dehydrogenase (17ß-HSD) activities and serum testosterone levels in rats' testicular tissues. Treatment with caffeine increased 3ß-HSD and 17ß-HSD as well as testosterone levels. Caffeine also reversed sperm viability, sperm motility and sperm count in testicular tissues of scopolamine-treated rats. Furthermore, scopolamine-induced oxidative damage in rats' epididymal and testicular tissues via reduction of thiol and non-protein thiol content as well as increase in reactive oxygen species (ROS) and malondialdehyde (MDA) levels. Caffeine attenuated oxidative stress in testicular and epididymal tissues of rats treated with scopolamine via increase in non-protein and protein thiol levels with concomitant reduction in ROS and MDA levels. CONCLUSION: This study revealed that caffeine (5 and 25 mg/kg) improved sperm quality, increased steroidogenic enzyme activities and attenuated oxidative damage in testis and epididymis of rats treated with scopolamine.


Assuntos
Amnésia/tratamento farmacológico , Cafeína/farmacologia , Epididimo/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testosterona/metabolismo , 17-Hidroxiesteroide Desidrogenases/metabolismo , Amnésia/induzido quimicamente , Amnésia/metabolismo , Animais , Modelos Animais de Doenças , Epididimo/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Oxirredução/efeitos dos fármacos , Ratos , Ratos Wistar , Escopolamina/farmacologia , Motilidade Espermática/efeitos dos fármacos , Testículo/metabolismo
3.
J Steroid Biochem Mol Biol ; 194: 105433, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31376460

RESUMO

The steroid hormones not only exert various endocrine functions but also act as the autocrine or paracrine factors in different tissues of mammals. In the present study, the seasonal expressions of androgen receptor (AR), estrogen receptors alpha and beta (ERα and ERß), aromatase cytochrome P450 (P450arom) and 5α-reductase 1, 2 were investigated in the epididymis of the muskrat. HE staining showed enlarged lumen and abundant sperm in the breeding season while reduced lumen with no sperm in the non-breeding season. The staining of AR was presented in nuclei of epithelial cells of the epididymis in both seasons. The immunostaining of ERα was localized in both nuclei and cytoplasm of epithelial cells of the epididymis during the breeding season, while the weak staining of ERα was only in the nuclei of epithelial cells during the non-breeding season. In contrast, ERß signal was negative in the epididymis of the muskrat in both seasons. The positive signals for P450arom and 5α-reductase 1, 2 were found in the cytoplasm of epithelial and smooth muscle cells during both seasons. Moreover, the protein and mRNA expression levels of AR, ERα, P450arom and 5α-reductase 1, 2 were significantly higher in the epididymis during the breeding season than those of the non-breeding season, and the expression level of 5α-reductase 1 was higher when compared with 5α-reductase 2. In addition, the levels of testosterone (T) and dihydrotestosterone (DHT) in the epididymis and serum were remarkably higher during the breeding season. Taken together, these findings suggested androgen and estrogen might play an important endocrine or autocrine/paracrine role to regulate the epididymal functions of the muskrat.


Assuntos
Aromatase/metabolismo , Colestenona 5 alfa-Redutase/metabolismo , Epididimo/metabolismo , Receptores Androgênicos/metabolismo , Receptores Estrogênicos/metabolismo , Animais , Aromatase/genética , Arvicolinae , Colestenona 5 alfa-Redutase/genética , Masculino , RNA Mensageiro/metabolismo , Receptores Androgênicos/genética , Receptores Estrogênicos/genética , Reprodução , Estações do Ano
4.
Food Chem Toxicol ; 133: 110780, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31449894

RESUMO

Dietary Apigenin (AP), a natural flavonoid from plants, could alleviate high-fat diet (HFD) induced obesity and its complication. Nonetheless, the direct correlation between dietary AP and their effects in adipose tissues remained unclear. In this study, male C57BL/6 mice were fed with low-fat diet, HFD with or without 0.04% (w/w) AP for 12 weeks. Dietary AP ameliorated HFD induced body weight gain, glucose intolerance, and insulin resistance. Energy expenditure was increased with no influence on energy intake, which indicated us that AP prevented obesity by enhancing energy export. Interestingly, AP activated lipolysis (ATGL/FOXO1/SIRT1) without higher cycling free fatty acids (FFAs). FFAs were consumed by the upregulation of fatty acid oxidation (AMPK/ACC), thermogenesis, and browning (UCP-1, PGC-1α). Additionally, adipose tissue metabolic inflammation (NF-кB, MAPK) was also reduced by AP. Our study proposed that dietary AP could be explored as a new dietary strategy to combat obesity and related insulin resistance.


Assuntos
Tecido Adiposo Marrom/efeitos dos fármacos , Apigenina/farmacologia , Lipólise/efeitos dos fármacos , Termogênese/efeitos dos fármacos , Tecido Adiposo Marrom/citologia , Tecido Adiposo Marrom/metabolismo , Animais , Dieta Hiperlipídica , Epididimo/citologia , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Intolerância à Glucose/dietoterapia , Resistência à Insulina , Masculino , Camundongos Endogâmicos C57BL , Gordura Subcutânea/citologia , Gordura Subcutânea/efeitos dos fármacos , Gordura Subcutânea/metabolismo , Ganho de Peso/efeitos dos fármacos
5.
J Infect Chemother ; 25(10): 832-834, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31255523

RESUMO

OBJECTIVES: Chlamydia trachomatis is one of the major pathogens causing acute epididymitis. Azithromycin (AZM) has a good efficacy against C. trachomatis; however, the ability of AZM to penetrate into human epididymal tissue has not yet been fully elucidated. Here, we examined the appropriate dosage of oral AZM for human epididymal tissue by site-specific pharmacokinetic/pharmacodynamic (PK/PD) analysis. METHODS: Patients with prostate cancer who underwent orchiectomy were included in this study. All patients received a 1-g dose of AZM before orchiectomy. Both epididymal tissue and blood samples were collected during surgery, and the drug concentrations were measured by high-performance liquid chromatography. All concentration-time data were analyzed with a three-compartment model with first-order absorption and elimination processes to simulate AZM concentrations in serum and epididymal tissue. RESULTS: A total of 10 patients were enrolled in the current study. For the observed values, the ratio of the epididymal concentration to the serum concentration was 5.13 ± 3.71 (mean ± standard deviation). For the simulated values, the maximum concentrations were 0.64 µg/mL at 2.42 h in serum and 1.96 µg/g at 4.10 h in epididymal tissue. The 24-h concentrations were 0.239 µg/mL in serum and 0.795 µg/g in epididymal tissue. CONCLUSIONS: The penetration of oral AZM into human epididymal tissue was examined to assess the potential application of AZM for the treatment of acute epididymitis. Based on the previous reports mentioning drug-susceptibility of C. trachomatis, multiple doses of oral AZM 1 g would be recommended for epididymitis based on the site-specific PK/PD.


Assuntos
Antibacterianos/farmacocinética , Azitromicina/farmacocinética , Epididimo/metabolismo , Administração Oral , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/administração & dosagem , Azitromicina/administração & dosagem , Infecções por Chlamydia/tratamento farmacológico , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/efeitos dos fármacos , Chlamydia trachomatis/patogenicidade , Relação Dose-Resposta a Droga , Farmacorresistência Bacteriana , Epididimo/microbiologia , Epididimite/tratamento farmacológico , Epididimite/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Orquiectomia , Neoplasias da Próstata/cirurgia , Distribuição Tecidual
6.
Exp Mol Pathol ; 110: 104271, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31251898

RESUMO

Testicular torsion/detorsion (T/D) is an inflammatory problem in men genital system with infertility effects. Cyclosporine A (CsA) as an immunosuppressant medication, exerts anti-inflammatory properties in tissue injuries. We sought to compare the efficacy of 3 doses of CsA on oxidative stress, apoptosis and epididymal sperm quality after ipsilateral testicular T/D. METHODS: 96 mature male rats were divided into six groups 16 each in: Control group (Group1), Sham operated (Group2), In rest groups, the right testis was twisted 720° in a clockwise direction for 1 h; T/D + 0.1% dimethylsulfoxide) DMSO((Group3), and in groups 4-6; CsA were administered 1, 5, and 10 mg/kg, intravenously (iv) 30 and 90 min after torsion, respectively. RESULTS: Tissue malondialdehyde (MDA) level and caspase-3 activity increased and catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities decreased in compared with control group 4 h after detorsion (p < .001). In six rats of each group 24 h after detorsion, histopathological changes and germ cell apoptosis were significantly deteriorated by determining mean of seminiferous tubules diameters (MSTD) and TUNEL assay. Moreover, 30 days after T/D, sperm concentration and motility were examined in rest of animals. CONCLUSIONS: Pre- and post-reperfusion CsA diminished MDA and caspase-3levels and normalized antioxidant enzymes activities. Germ cell apoptosis was significantly reduced, as well as, MSTD and long-term sperm insults were improved. Inhibition of mitochondrial permeability transition pore opening is suggested mechanism for cell protection against testicular T/D insults.


Assuntos
Apoptose/efeitos dos fármacos , Ciclosporina/farmacologia , Epididimo/efeitos dos fármacos , Traumatismo por Reperfusão/fisiopatologia , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Caspase 3/metabolismo , Catalase/metabolismo , Epididimo/metabolismo , Epididimo/patologia , Células Germinativas , Glutationa Peroxidase/metabolismo , Imunossupressores/farmacologia , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Torção do Cordão Espermático/fisiopatologia , Espermatozoides/patologia , Superóxido Dismutase/metabolismo , Testículo/irrigação sanguínea , Testículo/metabolismo
7.
Gen Comp Endocrinol ; 281: 153-163, 2019 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-31181195

RESUMO

Uroplakins (UPKs) play an important role in the normal and pathophysiology of the urothelium. They protect the urothelium and play a crucial role during urothelial infections by Uropathogenic E. coli. However, their functions beyond this organ system remain unexplored. A wide variety of proteins secreted in the male reproductive tract tissues contribute to spermatogenesis, sperm maturation, fertilization and innate immunity. However, the presence of UPKs and their possible contribution to the male reproductive tract physiology is not yet reported. Hence, in this study, we characterized UPKs in the male reproductive tract of rats. To the best of our knowledge, for the first time, we report the expression of UPKs in the male reproductive system. Upk1a, Upk1b, Upk2 and Upk3b mRNA and their corresponding proteins were abundantly expressed in the caput, cauda, testis, seminal vesicles and the prostate. Their expression was not developmentally regulated. UPK protein expression was also localized on the spermatozoa, suggesting a role for these proteins in sperm function. To study the role of UPKs in innate immunity, Upk mRNA expression in response to endotoxin challenge was evaluated in vitro and in vivo. In the rat testicular and epididymal cell lines, Upk mRNA levels increased in response to lipopolysaccharide challenge. However, in the caput, cauda, testes, seminal vesicle and prostate obtained from LPS treated rats, Upk mRNA expression was significantly reduced. Results of this study indicate a role for UPKs in male reproductive physiology and innate immune responses.


Assuntos
Genitália Masculina/metabolismo , Uroplaquinas/genética , Animais , Simulação por Computador , Endotoxinas/toxicidade , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Genitália Masculina/efeitos dos fármacos , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Wistar , Testículo/efeitos dos fármacos , Testículo/metabolismo , Distribuição Tecidual/efeitos dos fármacos , Uroplaquinas/metabolismo
8.
Environ Sci Pollut Res Int ; 26(23): 23306-23318, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31190303

RESUMO

The impact of Moringa oleifera leaf ethanol extract (MOLEE) was assessed on the expression of the steroidogenic genes (steroidogenic acute regulatory protein (StAR) and cytochrome P450c17 subfamily a (CYP17a) and luteinizing hormone receptor (LHR) gene) as well as on the cadmium chloride (CdCl2)-induced reproductive toxicity for 56 days in male rats. Four groups were used: control, Moringa-treated (MOLEE), CdCl2-treated, and CdCl2 + MOLEE groups. The reproductive toxicity of CdCl2 was confirmed; it caused a significant decrease in the accessory sex organ weights, testosterone level, testicular GST level, elevated MDA level (lipid peroxidation indicator), and histopathological alterations in seminiferous tubules, prostate, seminal vesicles, and epididymis as well as sperm characteristics. It also induced downregulation in the expression of StAR and CYP17a genes without change in the expression LHR gene. Eleven active compounds were detected in the GC-MS analysis of MOLEE; six of them have antioxidant properties, and five new compounds presented variable activities. MOLEE alone induced a stimulatory effect on the expression of steroidogenic and LHR genes. It restored the weight of reproductive organs to the control level; however, the recovery in sperm count, motility, abnormalities, percentage of alive sperm, testosterone, and MDA level are still comparable with the control level. Similar findings were also reported at the histological structure of the testes, epididymis, and accessory sex glands. Complete recovery of the GST enzyme activity was observed. Additionally, a restoration in the expression level of the steroidogenic genes was also reported. Our results indicated that the concurrent administration of MOLEE with CdCl2 can partially mitigate its harmful effects on male fertility.


Assuntos
Cloreto de Cádmio/toxicidade , Moringa/química , Fosfoproteínas/genética , Extratos Vegetais/farmacologia , Reprodução/efeitos dos fármacos , Esteroide 17-alfa-Hidroxilase/genética , Animais , Regulação para Baixo , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Expressão Gênica/efeitos dos fármacos , Masculino , Folhas de Planta/química , Ratos , Ratos Wistar , Reprodução/genética , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismo
9.
Toxicol Lett ; 311: 66-79, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31039416

RESUMO

Decabromodiphenyl ether (BDE-209), a flame retardant, interferes with thyroid homeostasis and androgen biosynthesis. BDE-209 evokes hyperglycemia through impaired glucose homeostasis in rat liver. This study is in continuation to our earlier work for a better understanding of whether or not BDE-209 affects testicular and epididymal physiology in relation to oxidative status in peripupertal mice offspring. Lactating female Parkes mice were orally gavaged with 500 and 700 mg/kg body weight of BDE-209 in corn oil from postnatal day (PND) 1 to PND 28. Male pups of lactating dams were sacrificed at PND 42. Maternal BDE-209 exposure during lactation increased apoptosis and oxidative status with altered expressions of various cell survival (Bcl-2), apoptotic (Bax and caspase-3) and oxidative stress (Nrf2 and HO-1) markers in testes and epididymis of peripubertal mice offspring. Testicular glucose and lactate concentrations were markedly reduced in these pups with down-regulation in GLUT3 and GLUT8 expressions and decreased LDH activity. Maternal BDE-209 exposure markedly affected fertility potential, epididymal histology, sialic acid concentration and sperm quality with decreased expression of epididymal Cx43 and AR in these mice offspring. Results thus suggest that maternal BDE-209 exposure during lactation causes reproductive toxicity in peripubertal mice offspring.


Assuntos
Epididimo/efeitos dos fármacos , Fertilidade/efeitos dos fármacos , Éteres Difenil Halogenados/toxicidade , Infertilidade Masculina/induzido quimicamente , Lactação , Estresse Oxidativo/efeitos dos fármacos , Testículo/efeitos dos fármacos , Fatores Etários , Animais , Conexina 43/metabolismo , Epididimo/metabolismo , Epididimo/patologia , Feminino , Glucose/metabolismo , Heme Oxigenase-1/metabolismo , Homeostase , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Infertilidade Masculina/fisiopatologia , Masculino , Exposição Materna , Proteínas de Membrana/metabolismo , Camundongos , Fator 2 Relacionado a NF-E2/metabolismo , Gravidez , Receptores Androgênicos/metabolismo , Desenvolvimento Sexual , Transdução de Sinais/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Espermatozoides/patologia , Testículo/metabolismo , Testículo/patologia
10.
Food Funct ; 10(6): 3324-3333, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31095144

RESUMO

The aim of the present study was to assess whether dietary advanced glycation end products (AGEs) induce testicular dysfunction. Using a BALB/c mouse model, AGE intake and serum levels were found to increase in AGE diet-treated mice relative to the controls. Histopathological damage was detected in the testes and epididymides of the AGE diet-induced mice. The total number of epididymal sperm decreased, and increased abnormal sperm rate was found in the mice. Moreover, the mice testes showed an increased level of the receptor for AGEs (RAGE) and malondialdehyde (MDA). Using a Sprague-Dawley rat model, AGE diet-induced rats showed 3- to 4-fold higher AGE intake than the controls. In these rats, higher serum and sperm MDA levels, decreased epididymal sperm numbers, and increased abnormal sperm rates were also observed. Silymarin, a natural AGE inhibitor, was found to restore these AGE-induced phenomena. Concluding from the above findings, dietary AGEs may promote testicular dysfunction.


Assuntos
Epididimo/citologia , Produtos Finais de Glicação Avançada/metabolismo , Espermatogênese , Espermatozoides/citologia , Testículo/citologia , Animais , Epididimo/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Sprague-Dawley , Receptor para Produtos Finais de Glicação Avançada/genética , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Contagem de Espermatozoides , Espermatozoides/metabolismo , Testículo/metabolismo
11.
Glycobiology ; 29(8): 593-607, 2019 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-31091305

RESUMO

Discoveries on involvement of glycan-protein recognition in many (patho)physiological processes are directing attention to exploring the significance of a fundamental structural aspect of sugar receptors beyond glycan specificity, i.e., occurrence of distinct types of modular architecture. In order to trace clues for defining design-functionality relationships in human lectins, a lectin's structural unit has been used as source material for engineering custom-made variants of the wild-type protein. Their availability facilitates comparative analysis toward the stated aim. With adhesion/growth-regulatory human galectin-1 as example, the strategy of evaluating how changes of its design (here, from the homodimer of non-covalently associated domains to (i) linker-connected di- and tetramers and (ii) a galectin-3-like protein) affect activity is illustrated by using three assay systems of increasing degree of glycan complexity. Whereas calorimetry with two cognate disaccharides and array testing with 647 (glyco)compounds disclosed no major changes, galectin histochemical staining profiles of tissue sections that present natural glycome complexity revealed differences between wild-type and linker-connected homo-oligomers as well as between the galectin-3-like variant and wild-type galectin-3 for cell-type positivity, level of intensity at the same site and susceptibility for inhibition by a bivalent glycocompound. These results underscore the strength of the documented approach. Moreover, they give direction to proceed to (i) extending its application to other members of this lectin family, especially galectin-3 and (ii) then analyzing impact of architectural alterations on cell surface lattice formation and ensuing biosignaling systematically, considering the variants' potential for translational medicine.


Assuntos
Galectina 1/metabolismo , Processamento de Proteína Pós-Traducional , Amino Açúcares/metabolismo , Animais , Sítios de Ligação , Epididimo/metabolismo , Galectina 1/química , Humanos , Jejuno/metabolismo , Lactose/análogos & derivados , Lactose/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ligação Proteica , Multimerização Proteica
12.
J Agric Food Chem ; 67(18): 5240-5249, 2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31008594

RESUMO

Fluoride is a widespread environmental pollutant that can induce low sperm quality and fertilizing ability; however, the underlying mechanism still remains unclear. Hence, we aimed to investigate the influence of fluoride on the sperm fertilizing ability via some key proteins in the epididymis. For this, 40 adult rats were assigned randomly into four groups. The control group was given distilled water, while the other three groups were given 25, 50, and 100 mg of NaF/L via drinking water for 56 days, respectively. After 1 day, epididymides were processed for sperm-egg binding, RNA extraction, western blot, and immunofluorescence analysis. Fluoride exposure reduced the ability of sperm to break down the egg cumulus cell layer. A further study revealed that fluoride altered the expression levels of genes and proteins related to acrosome reaction in vivo, including SPAM1, ACR, and PRSS21. However, fluoride only affected the expression of the ACR protein only in the epididymis but not in the testis. Fluoride also affected the expression levels of the membrane proteins CD9 and CD81 of epididymosomes in the epididymis. From the results, it can be concluded that fluoride exposure reduced the ability of sperm to break down the egg cumulus cell layer, which could be one of the reasons for decreased fertility ability in males treated with fluoride. These results provide some theoretical guidance and new ideas for treatments of low fertility, infertility, and other reproductive diseases.


Assuntos
Acrosina/metabolismo , Moléculas de Adesão Celular/metabolismo , Epididimo/efeitos dos fármacos , Fluoretos/farmacologia , Hialuronoglucosaminidase/metabolismo , Serina Endopeptidases/metabolismo , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Acrosina/genética , Animais , Moléculas de Adesão Celular/genética , Epididimo/metabolismo , Fertilização/efeitos dos fármacos , Hialuronoglucosaminidase/genética , Masculino , Ratos , Ratos Sprague-Dawley , Serina Endopeptidases/genética , Testículo/efeitos dos fármacos , Testículo/metabolismo
13.
Reprod Domest Anim ; 54(7): 956-963, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31004535

RESUMO

Cryptorchidism is associated with changes in the gonads and the spermatic duct system, which may cause infertility problems. Urocortin (UCN) is a corticotrophin-releasing hormone (CRH)-related peptide, which affects several functions of male genital organs. The aim of the present study was to investigate the expression of UCN and its receptors CRHR1 and CRHR2 using immunohistochemistry, western blotting and real-time reverse transcription polymerase chain reaction in tissues collected from the epididymis of normal and cryptorchid dogs. The lumen of the cryptic epididymal duct was found to be relatively smaller than that of the normal one, and interstitial tissue was abundant in the cryptic epididymis. In addition, only a few spermatids were observed in the lumen of the epididymal duct. Results showed that UCN, CRHR2 and CRHR1 were expressed in tissues collected from normal and cryptic epididymal ducts. Urocortin- and CRHR2-immunoreactivities (IRs) were detected in the principal cells of the caput, corpus and cauda of the normal and cryptic epididymides. CRHR1-IR was detected in vascular smooth muscles and fibromuscular cells surrounding epididymal tubules of the normal and cryptorchid dogs. Expression levels of UCN and CRHR2 mRNA were higher in cryptic epididymal ducts than that in normal epididymal ducts. These results suggest that UCN and its receptors might play a role in regulating the maturation and storage of spermatozoa. These findings indicated that the expression of these proteins could be modulated by the cryptorchidism condition.


Assuntos
Criptorquidismo/veterinária , Doenças do Cão/patologia , Cães , Epididimo/metabolismo , Urocortinas/metabolismo , Animais , Epididimo/anormalidades , Masculino , RNA Mensageiro , Receptores de Hormônio Liberador da Corticotropina/genética , Espermátides , Distribuição Tecidual , Urocortinas/genética
14.
Toxicol Lett ; 310: 1-6, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30980909

RESUMO

The increased number of cell divisions undergone by spermatogonia of older fathers cannot fully account for the observed increase in germline genetic damage. Studies have shown that the mechanisms induced in germ cells in response to oxidative damage varies with age, that DNA repair efficiency declines, and both sperm DNA damage and spontaneous mutations increase. However, it is not known whether the altered response with age is a cause, or consequence, of an age-associated change in cell susceptibility to genetic damage. Following a single 150 mg/kg dose of cyclophosphamide (CP), young (8-weeks old) and aged (17-month old) male mice were examined 24 h later for induced genetic damage in epididymal spermatozoa using the alkaline comet and sperm chromatin stability assays. Apoptosis among testicular cells was examined on tissue cross-sections using the TUNEL assay. Sperm showed no significant increase in DNA strand breaks with age (detected by the comet assay) and no change in sperm chromatin stability (detected by the SCSA assay). Following CP treatment, there was no effect on DNA-strand breakage but sperm chromatin instability was significantly higher. Furthermore, it was also significantly elevated in old treated, compared with young treated, animals suggesting that increased age affects the sensitivity of epididymal sperm to chromatin damage. There was no difference in apoptosis in testicular germ cells from either young or old control animals, while CP administration resulted in a significant increase in apoptosis among young animals but not old animals. Following genotoxin exposure, an increase in chromatin instability in the spermatozoa of old animals and a decrease in the ability of their testicular germ cells undergo apoptosis suggests an age-related decrease in genome protection mechanisms. Since those germ cells are only transiently present in the testis, it is likely that this age-related deterioration originates in the spermatogonial stem cells. The findings are also evidence that the safety evaluation of reproductive genotoxins should consider young and old individuals separately.


Assuntos
Ciclofosfamida/toxicidade , Epididimo/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Fatores Etários , Animais , Apoptose/efeitos dos fármacos , Montagem e Desmontagem da Cromatina/efeitos dos fármacos , Ensaio Cometa , Dano ao DNA , Epididimo/metabolismo , Epididimo/patologia , Marcação In Situ das Extremidades Cortadas , Masculino , Camundongos Endogâmicos C3H , Medição de Risco , Espermatozoides/metabolismo , Espermatozoides/patologia , Testículo/metabolismo , Testículo/patologia
15.
Int J Mol Sci ; 20(7)2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30978930

RESUMO

Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.


Assuntos
Criopreservação/veterinária , Preservação do Sêmen/veterinária , Espermatozoides/citologia , Suínos , Animais , Criopreservação/métodos , Ejaculação , Epididimo/citologia , Epididimo/metabolismo , Fertilização In Vitro , Masculino , Proteoma/metabolismo , Preservação do Sêmen/métodos , Motilidade Espermática , Espermatozoides/metabolismo , Suínos/fisiologia
16.
Genes Genomics ; 41(7): 757-766, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30859493

RESUMO

BACKGROUND: In the epididymis of bilateral castrated male rat, gene expression profile changed significantly. However, up to date, no study has investigated how these genes were regulated by microRNAs (miRNAs). OBJECTIVE: We investigated the alterations in the miRNA signature of the epididymis from sham-operated and bilaterally castrated rats. METHODS: By employing deep sequencing technology and qPCR, the global alterations of epididymal miRNA signature between sham-operated (Con-EP library) and bilaterally castrated rats (Cas-EP library) were explored. MiRNA-target interaction networks were annotated by GO and KEGG enrichment. RESULTS: We identified 313 and 306 known miRNAs as well as 152 and 114 novel miRNAs in the Con-EP and Cas-EP libraries, respectively. 59 miRNAs were differentially expressed, including 24 up-regulated and 35 down-regulated miRNAs, among which two up-regulated and three down-regulated ones were validated using qPCR. The expression of these miRNAs in the epididymides of rats at different postnatal ages showed regular changes from birth to adult, suggesting they were androgen-regulated. GO analysis showed that many of the miRNA targets were enriched in metabolic processes. KEGG analysis demonstrated that the targets mainly participated in the mitogen-activated protein kinase (MAPK) pathway. Moreover, 3 and 6 functional modules were detected among the up- and down-regulated miRNA target interaction networks, respectively, and these modules were involved in various biological processes. CONCLUSION: This study represents the first systematic investigation of alterations in the miRNA signature of the epididymis from bilaterally castrated rats and will provide useful resources for functional studies of the miRNAs in the male reproductive system.


Assuntos
Castração/efeitos adversos , Epididimo/metabolismo , MicroRNAs/genética , Transcriptoma , Animais , Epididimo/crescimento & desenvolvimento , Masculino , MicroRNAs/metabolismo , Ratos , Ratos Sprague-Dawley
17.
Cell Physiol Biochem ; 52(3): 421-434, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30845381

RESUMO

BACKGROUND/AIMS: The aim of this study was to evaluate the potential and significant applications of Sertoli cells (SCs) transplantation, and to explore the effect of transplantation on spermatogenesis process, in azospermic mice. METHODS: In this study, we utilized 18 adult mice (28‒30 g), divided into four experimental groups: (1) control, (2) vehicle (DMSO 2%) (10 µl) (3) busulfan and (4) busulfan+ SCs (1×104 cells/µL). SCs were isolated from the testis of 4-week-old mouse and after using anesthetics, 10 µl of SCs suspension (1×104 cells/µL) was injected over 3-5 min, into each testis and subsequently, sperm samples were collected from the tail of the epididymis. Afterward, the animals were euthanized and testis samples were taken for histopathology experiments, and RNA extraction, in order to examine the expression of c-kit, STRA8 and PCNA genes. RESULTS: Our data showed that SCs transplantation could notably increase the total sperm count and the number of testicular cells, such as spermatogonia, primary spermatocyte, round spermatid, SCs and Leydig cells, compared to the control, DMSO and busulfan groups. Furthermore, the result showed that the expression of c-kit and STRA8 were significantly decreased in busulfan and busulfan/SCs groups, at 8 weeks after the last injection (p<0.001), but no significant decrease was found for PCNA, compared to the control and DMSO groups (P<0.05). CONCLUSION: These findings suggest that SCs transplantation may be beneficial as a practical approach for therapeutic strategies in reproductive and regenerative medicine. We further highlighted the essential applications that might provide a mechanism for correcting fertility in males, suffering from cell deformity.


Assuntos
Células de Sertoli/transplante , Espermatogênese , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Bussulfano/farmacologia , Epididimo/citologia , Epididimo/metabolismo , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/metabolismo , Masculino , Camundongos , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/metabolismo , Medicina Regenerativa , Células de Sertoli/citologia , Motilidade Espermática , Espermátides/citologia , Espermátides/metabolismo , Espermatogênese/efeitos dos fármacos , Espermatogônias/citologia , Espermatogônias/fisiologia , Testículo/metabolismo , Testículo/patologia
18.
Nat Commun ; 10(1): 777, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30770815

RESUMO

Androgen receptor (AR) is regulated by SUMOylation at its transactivation domain. In vitro, the SUMOylation is linked to transcriptional repression and/or target gene-selective regulation. Here, we generated a mouse model (ArKI) in which the conserved SUMO acceptor lysines of AR are permanently abolished (ArK381R, K500R). ArKI males develop normally, without apparent defects in their systemic androgen action in reproductive tissues. However, the ArKI males are infertile. Their spermatogenesis appears unaffected, but their epididymal sperm maturation is defective, shown by severely compromised motility and fertilization capacity of the sperm. Fittingly, their epididymal AR chromatin-binding and gene expression associated with sperm maturation and function are misregulated. AR is SUMOylated in the wild-type epididymis but not in the testis, which could explain the tissue-specific response to the lack of AR SUMOylation. Our studies thus indicate that epididymal AR SUMOylation is essential for the post-testicular sperm maturation and normal reproductive capability of male mice.


Assuntos
Epididimo/metabolismo , Epididimo/fisiopatologia , Infertilidade Masculina/metabolismo , Infertilidade Masculina/fisiopatologia , Receptores Androgênicos/metabolismo , Espermatogênese/fisiologia , Animais , Epididimo/patologia , Humanos , Infertilidade Masculina/patologia , Masculino , Camundongos , Receptores Androgênicos/genética , Espermatogênese/genética , Sumoilação/genética , Sumoilação/fisiologia
19.
Toxicology ; 416: 44-53, 2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30721722

RESUMO

Imipenem is a beta-Lactam antibiotic characterized by a broad spectrum of activity. It is prescribed to treat severe infections. Our goal is to investigate toxicity induced in male rat reproductive systems following exposure to this drug (15, 50 or 100 mg/kg) compared to gentamicin (50 mg/kg) treatment. Effects of imipenem on reproductive organ weights, histoarchitecture, sperm parameters, and oxidative stress parameters were evaluated. Serum testosterone levels were measured. Apoptosis and inflammatory behaviors were investigated by immunohistochemical proteins expression analysis of apoptosis regulator BAX (Bax), B-cell lymphoma 2 (Bcl-2), and interleukin-1 beta (IL-1 beta) in testis. Results showed a significant decrease in male fertility parameters including sperm count, sperm motility, reproductive organ weights and serum testosterone levels after imipenem administration as compared to the control and gentamicin treated groups. Increased sperm abnormality was significant in animals treated with high doses of imipenem. Oxidative stress analysis revealed an expressed increase in lipid peroxidation and carbonyl groups levels in testicular tissues compared to control. Similar results were observed with superoxide dismutase and catalase activities from testicular tissues. In addition, severe testicular lesions were observed in the seminiferous tubules as well as important impairments in spermatogenesis testifying an inflammatory microenvironment confirmed by the intensive expression of IL1-beta and Bax protein by germinal cells and Bcl-2 by Leydig cells. In conclusion, imipenem treatment with high doses was found to lead to oxidative stress in male reproductive organs and an inflammatory microenvironment leading to spermatogenesis dysfunction and histopathological changes in the testis.


Assuntos
Antibacterianos/toxicidade , Microambiente Celular , Imipenem/toxicidade , Infertilidade Masculina/induzido quimicamente , Mediadores da Inflamação/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Epididimo/patologia , Gentamicinas/toxicidade , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Carbonilação Proteica/efeitos dos fármacos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Espermatozoides/patologia , Testículo/metabolismo , Testículo/patologia , Testosterona/sangue
20.
Theriogenology ; 128: 62-73, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30743105

RESUMO

Toll-like receptors (TLRs) are important molecules, which provide protection against infections of the reproductive tract. This study demonstrates for the first time the expression and localization patterns of TLRs in the caput, corpus and cauda segments of the epididymal duct (ED) and the vas deferens (VD) of adult domestic cats using immunohistochemistry and western blotting. While immunoblot analyses revealed relatively similar protein levels for TLRs 2, 4, 5, and 9 in three segments of the ED, the protein levels of TLR2 and TLR4 in the VD were found to be significantly higher than those measured in the ED segments (P < 0.05). On the other hand, immunostaining showed that TLRs exhibited regional- and cell-specific localization patterns. TLR2 and TLR5 were immunolocalized to the nucleus and cytoplasm of the principal cells in all ducts. TLR4 was restricted to the stereocilia, and TLR9 was located in the cytoplasm of the principal cells. Narrow cells displayed positive immunoreactions for TLR4 and TLR5. The basal cells of the different ED segments were positive for all four TLRs. TLR2, TLR5 and TLR9 were detected in the cytoplasmic droplets of the spermatozoa. TLR4 and TLR9 were detected along the entire length of the sperm tail, whilst TLR2 and TLR5 were absent in the midpiece. TLR2 and TLR5 were also detected in the equatorial segment of the sperm head. These results suggest that TLR2, TLR4, TLR5 and TLR9 are important not only for the protection of the ED, VD and spermatozoa but also for the maturation and storage of spermatozoa in the ED and VD, respectively.


Assuntos
Gatos/metabolismo , Epididimo/metabolismo , Receptores Toll-Like/metabolismo , Ducto Deferente/metabolismo , Animais , Western Blotting/veterinária , Gatos/crescimento & desenvolvimento , Imuno-Histoquímica/veterinária , Masculino , Receptor 2 Toll-Like/análise , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/análise , Receptor 4 Toll-Like/metabolismo , Receptor 5 Toll-Like/análise , Receptor 5 Toll-Like/metabolismo , Receptor Toll-Like 9/análise , Receptor Toll-Like 9/metabolismo , Receptores Toll-Like/análise
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