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1.
Chem Biol Interact ; 315: 108869, 2020 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-31682803

RESUMO

Spermatogenic dysfunction is one of the major secondary complications of male diabetes. Salidroside (SAL) is the important active ingredients isolated from Herba Cistanche, which exhibits numerous pharmacological activities such as antioxidant, anti-diabetic, and anti-inflammatory effects. The present study was designed to determine whether SAL contributes to the recovery from spermatogenic dysfunction in streptozotocin (STZ) induced type-1 diabetic mice. SAL (25, 50, or 100 mg/kg) and Clomiphene citrate (CC, 5 mg/kg) were orally administered to male type-1 diabetic mice for 10 weeks. Testis tissues were collected for histopathological and biochemical analysis. Moreover, reproductive organ weight, sperm parameters, and testicular cell DNA damage were estimated. The results revealed that SAL significantly improved the weight of the reproductive organs, sperm parameters and testicular morphology to different degrees in type-1 diabetic mice. Furthermore, reactive oxygen species (ROS) and malondialdehyde (MDA) levels were significantly reduced, and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), markedly increased in the testicular tissue after SAL treatment. In addition, our data also showed a marked downregulation the fluorescence expressions of p38 MAPK phosphorylation and upregulation the protein expressions of ZO-1, Occludin, Claudin-11 and N-cadherin after SAL administration (100 mg/kg) compared with the type-1 diabetic group. In conclusion, these results demonstrated that SAL exerts protective effects on type-1 diabetes-induced male spermatogenic dysfunction, which is likely mediated by inhibiting oxidative stress-mediated blood testis barrier damage.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 1/tratamento farmacológico , Glucosídeos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fenóis/farmacologia , Substâncias Protetoras/farmacologia , Espermatogênese/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Barreira Hematotesticular/metabolismo , Catalase/metabolismo , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/induzido quimicamente , Diabetes Mellitus Tipo 1/metabolismo , Glutationa/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Espécies Reativas de Oxigênio/metabolismo , Contagem de Espermatozoides/métodos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Estreptozocina/farmacologia , Superóxido Dismutase/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo
2.
Ecotoxicol Environ Saf ; 189: 110053, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31862514

RESUMO

Particulate matter with an aerodynamic diameter of less than 2.5 µm (PM2.5) derived from automobile exhaust can lead to serious male spermatogenesis dysfunction, but its specific molecular mechanism is unclear. In this experiment, we focused on the blood-testis barriers (BTB) and explored the intracellular mechanisms underlying the fertility toxicity of PM2.5 originating from automobile exhaust in the primary cultured Sertoli cells(SCs) of rats. After PM2.5 exposure, excessive reactive oxygen species (ROS) and increased apoptosis of SCs were detected. The expression of the BTB related proteins including ZO-1, Occludin, N-cadherin and ß-catenin were significantly decreased and the spatial arrangement of F-actin was completely disordered through Immunofluorescence and Western blots tests. The phosphorylation of Jun N-terminal kinase (JNK), extracellular signal regulatory kinase (ERK), p38 mitogen-activated protein kinase (MAPK) were upregulated and nuclear factor (erythroid-derived 2) -like 2-related factor (Nrf2) was downregulated respectively. However, combined utilization of vitamin C and E were observed to prevent the increase of ROS generation, reduce celluar apoptosis, increase the expression of BTB related proteins, reconstructed the spatial arrangement of F-actin as well as improved the Nrf2 expression and attenuated the phosphorylation of the MAPK kinases and cleaved caspase-3 levels. Furthermore, ERK inhibitor (SCH772984), JNK inhibitor (SP600125) and p38 MAPK inhibitor (SB203580) obviously up-regulated BTB-related proteins expression as well as activated Nrf2 expression at varying degrees, indicating that ROS-MAPKs-Nrf2 is involved in the signaling pathway that leads to PM2.5-induced spermatogenesis dysfunction. These findings indicate that PM2.5 derived from automobile exhaust causes oxidative stress, which in turn causes cellular apoptosis of SCs and damage of the blood-testis barrier, resulting male spermatogenesis dysfunction, in which ROS-MAPK-Nrf-2 pathways may play a key role.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Material Particulado/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Células de Sertoli/efeitos dos fármacos , Emissões de Veículos/toxicidade , Animais , Apoptose/efeitos dos fármacos , Barreira Hematotesticular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Ratos , Células de Sertoli/metabolismo , Células de Sertoli/patologia
3.
Ecotoxicol Environ Saf ; 187: 109824, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31654863

RESUMO

Microcystin-LR (MC-LR), a widespread environmental contaminant, has been shown to have potent acute testicular toxicity. However, magnitudes of toxic effects, induced by MCs, depend on route and magnitude of exposure to the toxin. In the present study, male mice were orally exposed 1, 10 or 100 µg/L MC-LR for 90 or 180 days, and pathological approach and the isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics were employed with testes. Proteomics revealed that a number of differentially altered proteins may be involved in MC-LR-induced chronic testicular toxicity. The biological process analysis indicated the altered proteins played an important role in biological adhesion, cellular process, response to stimulus or rhythmic process. The cellular component analysis revealed that most of the proteins with altered expression associated with cell part, extracellular region, extracellular region part, membrane, membrane part, organelle or organelle part. The molecular function showed that these proteins were critical in molecular transducer activity. Integrity analyses provide first compelling evidence that MC-LR significantly cause dysfunction of blood-testis barrier (BTB) through affecting tight junctions and gap junctions. Moreover, phosphatidylinositol 3-kinase (PI3K)/AKT eventually contributed to injury result from chronic low-level MC-LR treatment. Identification of proteins in testis responsive to MC-LR provides insights into molecular mechanisms of chronic toxicity of MCs.


Assuntos
Poluentes Ambientais/toxicidade , Microcistinas/toxicidade , Proteoma/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Barreira Hematotesticular/efeitos dos fármacos , Barreira Hematotesticular/fisiopatologia , Masculino , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Proteômica , Transdução de Sinais/efeitos dos fármacos , Testículo/metabolismo
4.
Chemosphere ; 237: 124410, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31362132

RESUMO

The profound influence of environmental chemicals on human health including inducing life-threatening gene mutation has been publicly recognized. Being a substitute for the extensively used endocrine-disrupting chemical BPA, Bisphenol AF (BPAF) has been known as teratogen with developmental toxicities and therefore potentially putting human into the risk of biological hazards. Herein, we deciphered the detrimental effects of BPAF on spermatogenesis and spermiotiliosis in sexual maturity of mice exposing to BPAF (5, 20, 50 mg/kg/d) for consecutive 28 days. BPAF exposure significantly compromises blood-testis barrier integrity and sperm quantity and quality in a dose-dependent manner. Sperms from BPAF exposure mice are featured by severe DNA damage, altered SUMOylation and ubiquitination dynamics and interfered epigenetic inheritance with hypermethylation of H3K27me3 presumably due to the aggregation of cellular reactive oxygen species (ROS). Furthermore, BPAF treatment (50 µM for 24 h) compromises cytoskeleton architecture and tight junction permeability in primary cultured Sertoli cells evidenced by dysfunction of actin regulatory proteins (e.g. Arp3 and Palladin) via activation of ERK signaling, thereby perturbing the privilege microenvironment created by Sertoli cells for spermatogenesis. Overall, our study determines BPAF is deleterious for male fertility, leading to a better appreciation of its toxicological features in our life.


Assuntos
Compostos Benzidrílicos/toxicidade , Barreira Hematotesticular/efeitos dos fármacos , Fenóis/toxicidade , Espermatozoides/efeitos dos fármacos , Animais , Compostos Benzidrílicos/administração & dosagem , Dano ao DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Disruptores Endócrinos/administração & dosagem , Disruptores Endócrinos/toxicidade , Epigênese Genética/efeitos dos fármacos , Histonas/metabolismo , Lisina/metabolismo , Masculino , Camundongos , Fenóis/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologia , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatozoides/metabolismo , Espermatozoides/patologia , Sumoilação/efeitos dos fármacos , Ubiquitinação/efeitos dos fármacos
5.
Nanotechnology ; 30(45): 455101, 2019 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-31362276

RESUMO

Zinc-based nanoparticles are promising materials for various applications, including in biomedicine. The aim of our study was to determine the effect of fluorescent europium-doped zinc oxide nanoparticles (ZnO:Eu NPs) on sperm parameters, cell apoptosis and integrity of the blood-testis barrier (BTB) in mice. Nanostructures were orally administered to adult mice (n = 34). Animals were sacrificed after 3 h, 24 h, 7 d and 14 d following oral administration. Sperm was collected and analysed for viability and kinetic parameters. Collected testes were quantitatively analysed for accumulation of ZnO:Eu NPs. Microscopic evaluation based on immunofluorescence and histopathological studies were also conducted. Results showed that ZnO:Eu NPs were able to overcome the BTB with their subsequent accumulation in the testis. No toxic or pro-apoptotic effects of nanoparticles on the male reproductive system were observed. The results suggested that ZnO:Eu NPs were able to accumulate in the testis with no negative impact on sperm parameters, tissue architecture or the integrity of the BTB.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Espermatozoides/citologia , Óxido de Zinco/administração & dosagem , Administração Oral , Animais , Apoptose , Európio/administração & dosagem , Európio/química , Masculino , Camundongos , Nanopartículas , Espermatozoides/efeitos dos fármacos , Óxido de Zinco/química , Óxido de Zinco/farmacologia
6.
Hum Exp Toxicol ; 38(12): 1329-1343, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31272229

RESUMO

Doxorubicin (DOX) is an anthracycline derivative antibiotic that still frequently used in the treatment of solid tumors and hematological malignancies. The clinical use of DOX is largely restricted due to acute and chronic renal, cardiac, hematological, and testicular toxicities. Previous studies have indicated that oxidative stress, lipid peroxidation, and apoptosis in germ cells are the main factors in DOX-induced testicular toxicity, but the entire molecular mechanisms that responsible for DOX-induced testicular damage are not yet fully understood. Fluvastatin is a cholesterol-lowering agent that acts by inhibiting hydroxylmethyl glutaryl coenzyme A, the key enzyme for cholesterol biosynthesis. In addition to its cholesterol-lowering effect, fluvastatin showed an antioxidant effect by cleaning hydroxyl and superoxide radicals and this drug could have a protective effect by acting on the mammalian target of rapamycin (mTOR) signal pathway in testicular damage caused by obesity. This study aimed to investigate the possible protective and therapeutic effects of fluvastatin on the DOX-induced testicular toxicity model by histochemical, immunohistochemical, biochemical, and real-time polymerase chain reaction analyses. The present study indicates that fluvastatin may have a protective and therapeutic effect by removing reactive oxygen species and by regulating the mTOR, connexin 43, and matrix metalloproteinase 9 protein and messenger ribonucleic acid expressions, which play an important role in regulating the blood-testis barrier. On the other hand, the use of fluvastatin as a protective/prophylactic agent was found to be more effective than the use of this drug for treatment. In light of this information, fluvastatin may be a candidate agent that can be used to prevent testicular toxicity observed in men receiving DOX treatment.


Assuntos
Antibióticos Antineoplásicos/efeitos adversos , Doxorrubicina/efeitos adversos , Fluvastatina/uso terapêutico , Substâncias Protetoras/uso terapêutico , Testículo/efeitos dos fármacos , Animais , Barreira Hematotesticular/efeitos dos fármacos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Contagem de Espermatozoides , Serina-Treonina Quinases TOR/metabolismo , Testículo/metabolismo , Testículo/patologia
7.
Andrologia ; 51(6): e13285, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31006889

RESUMO

The present study was designed to investigate the therapeutic effect of bone marrow MSC-derived factors on gonadotropic toxicity induced by busulfan in vivo. The conditioned media (CM) was obtained from MSCs in serum-free incubation for 48 hr and concentrated ~25-fold by ultrafiltration. The CM of HEK 293 cells was treated as control (293-CM). MSC-CM was injected into busulfan mice via caudal veins after 1 day of busulfan treatment for 2 weeks (200 µl per dose/twice weekly). Compared to the 293-CM group, testicular injury was delayed in MSC-CM group, including reduced vacuolations of cells in the basal compartment of the seminiferous epithelium and detachment of cells from basement membrane. Apoptotic spermatogenic cells were significantly decreased in MSC-CM group (p ï¼œ 0.05). Interesting N-cadherin,ICAM-1 and P-cadherin expressions significantly increased in MSC-CM group, while occludin, ZO-1 and connexin 43 expressions showed no difference among MSC-CM, 293-CM and busulfan groups. Present results suggest MSC-secreted factors protect spermatogenesis impairment after busulfan treatment by reducing the apoptosis of spermatogenic cells and enhancing intercellular adhesion molecule expressions.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Bussulfano/toxicidade , Meios de Cultivo Condicionados/farmacologia , Infertilidade Masculina/tratamento farmacológico , Células-Tronco Mesenquimais/metabolismo , Animais , Apoptose/efeitos dos fármacos , Barreira Hematotesticular/citologia , Barreira Hematotesticular/patologia , Caderinas/metabolismo , Adesão Celular/efeitos dos fármacos , Meios de Cultivo Condicionados/metabolismo , Modelos Animais de Doenças , Células HEK293 , Humanos , Infertilidade Masculina/induzido quimicamente , Infertilidade Masculina/patologia , Molécula 1 de Adesão Intercelular/metabolismo , Masculino , Camundongos , Espermatogênese/efeitos dos fármacos
8.
Mol Nutr Food Res ; 63(10): e1800843, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30924608

RESUMO

SCOPE: Luteolin, a natural flavonoid, displays protective activities to testicular tissue. However, the molecular mechanisms are still unclear. In this study, the aim is to identify the protective effects and underlying mechanisms of luteolin against triptolide (TP)-induced damage of testicular tissue. METHODS AND RESULTS: Pre-incubation of Sertoli cells (SCs) with luteolin results in a significant reduction of TP-induced apoptotic cells, which occurs concomitantly with the effective inhibition of reactive oxygen species accumulation. Luteolin results in a significant reduction in testicular damage and spermatogenesis dysfunction in a mouse model of testicular damage. Mechanistic studies reveal that luteolin significantly triggers Nrf2 translocation, increases antioxidant response element-luciferase reporter activity, and induces antioxidant enzyme expression. Nrf2 siRNA reduces luteolin-induced protection in SCs. Besides inhibiting apoptosis, luteolin recovers the blood-testis barrier (BTB) integrity by upregulating connexin43 (Cx43) expression. Moreover, specifically blocked Cx43 activity completely blocks repairmen of luteolin to BTB values. In accordance with in vitro results, luteolin suppresses testicular injury and spermatogenesis dysfunction by activation of Nrf2 and Cx43 in a testicular injury model. CONCLUSION: Luteolin is identified as a novel active ingredient that contributes to the protective activity in testicular damage through activating the Nrf2 signaling pathway and by upregulating Cx43.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Conexina 43/metabolismo , Luteolina/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Testículo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Barreira Hematotesticular/fisiologia , Diterpenos/toxicidade , Compostos de Epóxi/toxicidade , Masculino , Camundongos Endogâmicos ICR , Estresse Oxidativo/efeitos dos fármacos , Fenantrenos/toxicidade , Substâncias Protetoras/farmacologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologia , Transdução de Sinais/efeitos dos fármacos , Testículo/metabolismo , Testículo/patologia , Regulação para Cima/efeitos dos fármacos
9.
Andrologia ; 51(5): e13241, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30706522

RESUMO

Mirabegron is a selective beta3-adrenoceptor (ß3 -AR) agonist, which is commonly used for the treatment of overactive bladder. This medicine is associated with atrophy of reproductive organs in rats. However, no study has examined the detailed action and mechanism of its toxicity in reproductive cells. In this study, we examined the effect of mirabegron on primary cultured rat Sertoli cells. Firstly, RT-PCR and immunocytochemistry revealed that ß3 -AR was present in rat Sertoli cells. Then, primary cultured rat Sertoli cells were treated with mirabegron. Quantitative real-time PCR revealed that mirabegron treatment induced a significant increase in claudin-11 mRNA, which is crucial for spermatogenesis. Western blot analysis also showed that mirabegron treatment significantly activated p44/42 mitogen-activated protein kinase (MAPK). After additional treatment with U0126, a specific noncompetitive inhibitor of mitogen-activated protein kinase kinase (MAPKK), the upregulation of claudin-11 mRNA induced by mirabegron was reduced. At the same time, immunocytochemistry showed mirabegron treatment disturbed claudin-11 localisation to tight junction, which was recovered when treated with mirabegron in the presence of U0126. These results suggest that mirabegron treatment is associated with assembly of the blood-testis barrier through p44/42 MAPK pathway. These findings could explain one of the underlying mechanisms of reproductive toxicity induced by mirabegron.


Assuntos
Acetanilidas/toxicidade , Agonistas de Receptores Adrenérgicos beta 3/toxicidade , Barreira Hematotesticular/efeitos dos fármacos , Células de Sertoli/efeitos dos fármacos , Tiazóis/toxicidade , Junções Íntimas/efeitos dos fármacos , Animais , Butadienos/farmacologia , Células Cultivadas , Claudinas/metabolismo , Masculino , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Nitrilos/farmacologia , Cultura Primária de Células , Ratos , Ratos Sprague-Dawley , Células de Sertoli/citologia , Células de Sertoli/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Junções Íntimas/metabolismo
10.
Ecotoxicol Environ Saf ; 171: 475-483, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-30639874

RESUMO

Bisphenol A (BPA), an environmental contaminant, has been shown to disturb the dynamics of Sertoli cell blood-testis barrier (BTB) in mammal testis. However, the effects of BPA on Sertoli cell barrier (SC barrier) were little known in fish to date. To evaluate the potential mechanism of reproductive toxicity of BPA, we studied the damage of SC barrier using in vivo models. In this study, male adult rare minnow Gobiocypris rarus were exposed to 15 µg/L BPA for 7-35 days. Gonadal histology and the integrity of SC barrier were analyzed. Meanwhile, the expressions of SC barrier -associated proteins, tumor necrosis factor (TNFα) content, and the mRNA expressions of genes in the mitogen activated protein kinase (MAPK) pathway were detected. Histological analysis demonstrated 15 µg/L BPA promoted the infiltration of inflammatory cells in fish testes after 7-days exposure. The biotin tracer assay showed that 7-days BPA exposure increased permeability for spermatid cysts. In addition, the BPA treatment caused increased TNFα in testis, which was reportedly related to SC barrier impairment. The expressions of Occludin and ß-Catenin protein were significantly decreased in the testes after 7- and 21-days exposure. BPA also altered the mRNA expressions of occludin, ß-catenin, p38 MAPK and JNK. Therefore, the detrimental effects of BPA on reproduction of male fish may attribute to the disturbed expressions of SC junction proteins.


Assuntos
Compostos Benzidrílicos/toxicidade , Barreira Hematotesticular/efeitos dos fármacos , Cyprinidae , Fenóis/toxicidade , Células de Sertoli/efeitos dos fármacos , Animais , Barreira Hematotesticular/metabolismo , Gônadas/efeitos dos fármacos , Gônadas/metabolismo , Masculino , Ocludina/genética , Ocludina/metabolismo , Células de Sertoli/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
11.
Environ Sci Pollut Res Int ; 26(5): 4801-4820, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30565106

RESUMO

Deca-bromodiphenyl ether (BDE-209) regulates various aspects of spermatogenesis and male fertility through its effect on estrogen receptor α (ERα), but the underlying mechanism remains unclear. Because molecular mechanisms such as remodeling of the blood-testis barrier (BTB) play crucial roles in spermatogenesis, we investigated the disruptive effects of ERα agonists on the BTB in spermatogenesis. In this study, 0, 300, and 500 mg/kg/day of BDE-209 were administered to pregnant adult mice by oral gavage from gestation day 7 to postnatal day 21. SerW3 cells were treated with methylpiperidino pyrazole (MPP) for 30 min before being treated with 50 µg/mL of BDE-209. BDE-209 increases ERα in time- and dose-dependent manners and decreases formin 1 and BTB-associated protein in F1 male mice. Furthermore, BDE-209 impairs the structure and function of the BTB. Activation of ERα signaling could disrupt the BTB, leading to spermatogenesis dysfunction. The results identified the role of ERα in BTB disruption during spermatogenesis and suggested that BTB disruption occurs because of exposure to BDE-209, which could potentially affect spermatogenesis. In conclusion, Sertoli cells seem to be the primary target of BDE-209 in the perinatal period, and this period constitutes a critical window of susceptibility to BDE-209. Also, the SerW3 cell model may not be a particularly useful cell model for studying the function of the cytoskeleton.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Receptor alfa de Estrogênio/metabolismo , Éteres Difenil Halogenados/toxicidade , Células de Sertoli/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Animais , Barreira Hematotesticular/fisiologia , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Proteínas Fetais/metabolismo , Éteres Difenil Halogenados/administração & dosagem , Éteres Difenil Halogenados/farmacocinética , Masculino , Camundongos Endogâmicos ICR , Proteínas dos Microfilamentos/metabolismo , Proteínas Nucleares/metabolismo , Gravidez , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/fisiologia , Testículo/efeitos dos fármacos
12.
Toxicol Appl Pharmacol ; 360: 257-272, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30291936

RESUMO

Methamidophos (MET) is an organophosphate (OP) pesticide widely used in agriculture in developing countries. MET causes adverse effects in male reproductive function in humans and experimental animals, but the underlying mechanisms remain largely unknown. We explored the effect of MET on mice testes (5 mg/kg/day/4 days), finding that this pesticide opens the blood-testis barrier and perturbs spermatogenesis, generating the appearance of immature germ cells in the epididymis. In the seminiferous tubules, MET treatment changed the level of expression or modified the stage-specific localization of tight junction (TJ) proteins ZO-1, ZO-2, occludin, and claudin-3. In contrast, claudin-11 was barely altered. MET also modified the shape of claudin-11, and ZO-2 at the cell border, from a zigzag to a more linear pattern. In addition, MET diminished the expression of ZO-2 in spermatids present in seminiferous tubules, induced the phosphorylation of ZO-2 and occludin in testes and reduced the interaction between these proteins assessed by co-immunoprecipitation. MET formed covalent bonds with ZO-2 in serine, tyrosine and lysine residues. The covalent modifications formed on ZO-2 at putative phosphorylation sites might interfere with ZO-2 interaction with regulatory molecules and other TJ proteins. MET bonds formed at ZO-2 ubiquitination sites likely interfere with ZO-2 degradation and TJ sealing, based on results obtained in cultured epithelial cells transfected with ZO-2 mutated at a MET target lysine residue. Our results shed light on MET male reproductive toxicity and are important to improve regulations regarding the use of OP pesticides and to protect the health of agricultural workers.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Inseticidas/farmacologia , Organofosfatos/farmacologia , Compostos Organotiofosforados/farmacologia , Proteína da Zônula de Oclusão-2/metabolismo , Animais , Barreira Hematotesticular/metabolismo , Claudinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ocludina/metabolismo , Fosfoproteínas/metabolismo , Fosforilação/efeitos dos fármacos , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/metabolismo , Espermatogênese/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
13.
Environ Toxicol Pharmacol ; 63: 115-126, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30212741

RESUMO

Sertoli cells were treated with 0, 20, 40, 60 and 80 µg/L of MC-LR to investigate its toxic effects, mechanism of action and immune response of the cells. Our results revealed that treatment containing 20 µg/L of MC-LR was non-toxic to the cells. Treatments containing 40, 60 and 80 µg/L of MC-LR reduced the cell viability, induced nuclear morphological changes and downregulated the blood-testis barrier constituent proteins within 48 h after treatment. The toll-like receptor 4 (TLR4) and nuclear factor-kappaB (NF-kB) were activated and significantly (P < 0.05) upregulated in cells treated with 40, 60 and 80 µg/L of MC-LR compared to the control. The pro-inflammatory cytokines were upregulated within 48 h after treatment. However commencing from 72 h, upregulation of anti-inflammatory cytokines and expression of blood-testis barrier constituent proteins was observed. This study indicates that MC-LR induced inflammatory response in bovine Sertoli cell via activation of TLR4/NF-kB signaling pathway.


Assuntos
Microcistinas/toxicidade , Células de Sertoli/citologia , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima , Animais , Barreira Hematotesticular/efeitos dos fármacos , Bovinos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Testes de Toxicidade
14.
Ecotoxicol Environ Saf ; 166: 165-175, 2018 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-30267989

RESUMO

This study was conducted to investigate the ameliorative effect of selenium on microcystin-LR induced toxicity in bovine Sertoli cells. Bovine Sertoli cells were pretreated with selenium (Na2SeO3) for 24 h after which selenium pretreated and non-pretreated Sertoli cells were cultured in medium containing 10% heat activated fetal bovine serum FBS+ 80 µg/L MC-LR to assess its ameliorative effect on MC-LR toxicity. The results show that selenium pretreatment inhibited the MC-LR induced mitophagy, downregulation and mislocalization of blood-testis barrier constituent proteins in bovine Sertoli cells via NF-kB and cytochrome c release blockage. The observed downregulation of electron transport chain (ETC) related genes (mt-ND2, COX-1, COX-2) and upregulation of inflammatory cytokines (IL-6, TNF-α, IL-1ß, IFN-γ, IL-4, IL-10, 1 L-13, TGFß1) in non-pretreated cells exposed to MC-LR were ameliorated in selenium pretreated cells. There was no significant difference (P > 0.05) in the protein levels of blood-testis barrier constituent proteins (ZO-1, occludin, connexin-43, CTNNB1, N-cadherin) and mitochondria related genes (mt-ND2, COX-1, COX-2, ACAT1, mtTFA) of selenium pretreated Sertoli cell compared to the control. Taken together, we conclude that selenium inhibits MC-LR caused Mitophagy, downregulation and mislocalization of blood-testis barrier proteins of bovine Sertoli cell via mitochondrial and TLR4/NF-kB signaling pathways blockage.


Assuntos
Microcistinas/toxicidade , Mitocôndrias/efeitos dos fármacos , Células de Sertoli/efeitos dos fármacos , Selenito de Sódio/farmacologia , Animais , Barreira Hematotesticular/efeitos dos fármacos , Bovinos , Citocinas/metabolismo , Regulação para Baixo , Masculino , Mitocôndrias/metabolismo , NF-kappa B/antagonistas & inibidores , Células de Sertoli/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidores
15.
J Reprod Dev ; 64(6): 511-522, 2018 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-30175719

RESUMO

Stem cell homing is a complex phenomenon that involves multiple steps; thus far, attempts to increase homing efficiency have met with limited success. Spermatogonial stem cells (SSCs) migrate to the niche after microinjection into seminiferous tubules, but the homing efficiency is very low. Here we report that reversible disruption of the blood-testis barrier (BTB) between Sertoli cells enhances the homing efficiency of SSCs. We found that SSCs on a C57BL/6 background are triggered to proliferate in vitro when MHY1485, which stimulates MTORC, were added to culture medium. However, the cultured cells did not produce offspring by direct injection into the seminiferous tubules. When acyline, a gonadotropin-releasing hormone (GnRH) analogue, was administered into infertile recipients, SSC colonization increased by ~5-fold and the recipients sired offspring. In contrast, both untreated individuals and recipients that received leuprolide, another GnRH analogue, remained infertile. Acyline not only decreased CLDN5 expression but also impaired the BTB, suggesting that increased colonization was caused by efficient SSC migration through the BTB. Enhancement of stem cell homing by tight junction protein manipulation constitutes a new approach to improve homing efficiency, and similar strategy may be applicable to other self-renewing tissues.


Assuntos
Barreira Hematotesticular/metabolismo , Células de Sertoli/metabolismo , Espermatogônias/metabolismo , Testículo/metabolismo , Animais , Barreira Hematotesticular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Claudina-5/metabolismo , Masculino , Camundongos , Morfolinas/farmacologia , Oligopeptídeos/farmacologia , Células de Sertoli/citologia , Células de Sertoli/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatogênese/fisiologia , Espermatogônias/citologia , Espermatogônias/efeitos dos fármacos , Nicho de Células-Tronco/efeitos dos fármacos , Testículo/efeitos dos fármacos , Triazinas/farmacologia
16.
Chemosphere ; 211: 826-833, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30099167

RESUMO

Fluoride is known to affect the pro-inflammatory cytokines in the testis. Most of the recent literatures cited that cytokines regulate the blood-testis-barrier (BTB). However, the involvement of cytokines in the fluoride induced toxicity in BTB remains unclear. In order to study this, 60 male Sprague-Dawley (SD) rats were taken and randomly divided into 5 groups which included four fluoride groups exposed to 0, 25, 50, and 100 mg/L NaF in distilled water and one positive control group. On the 29th day of fluoride exposure, the positive control group rats were administered 0.1% CaCl2 solution. Biotin tracer technology and transmission electron microscopy (TEM) analysis were applied to evaluate the function and ultra-structure of BTB. The expression levels of the BTB associated proteins, actin relative protein 3 (Arp3), interleukin-1 alpha (IL-1α), and transforming growth factor beta-3 (TGF-ß3) were determined using Western blotting and Enzyme Linked Immunosorbent Assay (ELISA) respectively, meanwhile the actin filament (F-actin) was detected by fluorescent phalloidin conjugates. Our results revealed that the function and the ultra-structure of BTB in all the fluoride treated groups were damaged with a concomitant significant decreases in basal ectoplasmic specialization (basal ES), associated protein ß-catenin, and F-actin. Moreover, Arp3 levels were significantly increased in 50 and 100 mg/L NaF groups. Meanwhile, IL-1α significantly increased in all the fluoride treated groups. In summary, we concluded that an increase in IL-1α induced by NaF significantly decreased the expression of F-actin and the organization of F-actin highly branched, which might facilitate the BTB's functional and ultra-structural variations.


Assuntos
Actinas/metabolismo , Barreira Hematotesticular/efeitos dos fármacos , Fluoretos/toxicidade , Testículo/efeitos dos fármacos , Proteína 3 Relacionada a Actina/metabolismo , Animais , Relação Dose-Resposta a Droga , Interleucina-1alfa/biossíntese , Interleucina-1alfa/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Testículo/irrigação sanguínea , Testículo/metabolismo
17.
Toxicol Lett ; 295: 277-287, 2018 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-29981920

RESUMO

There are reports of fluorochloridone (FLC)-induced male reproductive toxicity, but the underlying toxicological mechanisms remain unknown. In this study, we looked at how FLC exposure affected the integrity of the blood-testis barrier (BTB) and the Sertoli cell barrier and studied the molecular mechanisms. Male rats received gavage administration of FLC (30 mg/kg/d) for 14 consecutive days with sample collection at the 7th and 14th day; and primary cultured Sertoli cells were treated with 0-10 µM FLC in vitro for 24 h. Our in vivo findings revealed that FLC exposure caused time-dependent testicular injuries, sperm quality decrease as well as adverse changes in BTB integrity, F-actin organization, and expressions of claudin-11 and Arp3. In Sertoli cells isolated from FLC-treated rat testis, Sertoli cell barrier tightness was increased. In Sertoli cells in vitro exposed to FLC, abnormal changes in the barrier permeability were also observed, and the protein expressions of occludin, claudin-11, ZO-1, connexin-43, and Arp3 were significantly decreased in a dose- and time-dependent manner. Furthermore, the FLC-induced adverse changes in Sertoli cell barrier and F-actin were partly alleviated by the induction of Arp3 overexpression. In conclusion, our findings revealed that FLC perturbed BTB/Sertoli cell barrier function through Arp3-mediated F-actin disorganization.


Assuntos
Citoesqueleto de Actina/efeitos dos fármacos , Proteína 3 Relacionada a Actina/metabolismo , Actinas/metabolismo , Poluentes Ocupacionais do Ar/toxicidade , Barreira Hematotesticular/efeitos dos fármacos , Pirrolidinonas/toxicidade , Reprodução/efeitos dos fármacos , Células de Sertoli/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/patologia , Proteína 3 Relacionada a Actina/genética , Animais , Barreira Hematotesticular/metabolismo , Barreira Hematotesticular/patologia , Células Cultivadas , Relação Dose-Resposta a Droga , Masculino , Permeabilidade , Ratos Sprague-Dawley , Medição de Risco , Células de Sertoli/metabolismo , Células de Sertoli/patologia , Transdução de Sinais/efeitos dos fármacos , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Junções Íntimas/patologia , Fatores de Tempo
18.
Toxicol Lett ; 296: 114-124, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30055240

RESUMO

Melamine (MA) exposure causes male reproductive toxicity, however, the mechanism remains unclear. In this study, we investigated the roles of the blood-testis barrier (BTB) in MA-induced reproductive toxicity in piglets. Male weaned piglets were exposed to MA concentrations of 0, 100, 300, and 1000 mg/kg in the diet for 10 weeks. They were euthanized on days 1, 7 and 14 after the final exposure. Body and organ weights, serum biochemistry and testosterone, gross and histopathological changes, and BTB ultrastructure and integrity were assessed. BTB junction protein expression levels and protein levels of the mitogen-activated protein kinase (MAPK) pathway in testes were measured. We found that MA dose-dependently decreased serum testosterone levels and caused gross and histopathological lesions in the testis and epididymis. Marked BTB damage was evidenced by abnormal changes in BTB ultrastructure and increased BTB permeability. Furthermore, MA decreased the protein levels of ZO-1, occludin, N-cadherin, and connexin-43, paralleled by increased the protein levels of p-Erk, p-JNK and p-p38 in testes, suggesting that MA disrupted BTB by downregulating the expressions of BTB junction proteins, with possible involvement of the MAPK signaling pathway. In conclusion, MA exposure gives rise to testicular toxicity in male piglets through destroying BTB integrity.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Doenças Testiculares/induzido quimicamente , Triazinas/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sus scrofa , Suínos , Doenças Testiculares/patologia , Testosterona/sangue
19.
Reprod Toxicol ; 81: 17-27, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29940330

RESUMO

As an environmental endocrine disruptor, Di-(2-ethylhexyl) phthalate (DEHP) affects blood-testis barrier (BTB)-associated proteins expression, which compromises BTB integrity and causes infertility. Notably, DEHP-induced testicular toxicity is related to oxidative stress, but the specific mechanism remains unclear. Therefore, we sought to investigate this mechanism and determine whether vitamin C and vitamin E administration would attenuate the BTB impairment induced by DEHP in vivo and by Mono-(2-Ethylhexyl) Phthalate (MEHP) in vitro, respectively. HE staining and EM found that DEHP exposure led to spermatogenesis dysfunction and BTB disruption, respectively. The Western blot and immunofluorescence results showed that DEHP exposure caused BTB impairment through oxidative stress-mediated p38 mitogen-activated protein kinase (MAPK) signaling pathway. Furthermore, Vitamin E and vitamin C could alleviate the oxidative stress, block DEHP-induced spermatogenesis dysfunction and BTB disruption by inhibiting p38 MAPK signaling pathway. In summary, vitamin E and vitamin C are good candidates for the treatment of DEHP-induced male infertility.


Assuntos
Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Barreira Hematotesticular/efeitos dos fármacos , Dietilexilftalato/toxicidade , Disruptores Endócrinos/toxicidade , Vitamina E/farmacologia , Vitaminas/farmacologia , Animais , Barreira Hematotesticular/metabolismo , Dietilexilftalato/análogos & derivados , Infertilidade Masculina/tratamento farmacológico , Masculino , Ratos Sprague-Dawley , Espermatogênese/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
20.
Reprod Biol Endocrinol ; 16(1): 55, 2018 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-29855380

RESUMO

BACKGROUND: Serum leptin levels are augmented in obese infertile men and in men with azoospermia. They also correlate inversely with sperm concentration, motility and normal forms. The mechanisms underlying the adverse effects of excess leptin on male reproductive function remain unclear. The present study aimed to evaluate the effects of exogenous leptin on sperm parameters in mice and to explore the underlying mechanisms. METHODS: We treated normal adult male mice with saline, 0.1, 0.5 or 3 mg/kg leptin daily for 2 weeks. After treatment, serum leptin levels, serum testosterone levels, sperm parameters and testicular cell apoptosis were evaluated. Blood testis barrier integrity and the expression of tight junction-associated proteins in testes were also assessed. We further verified the direct effects of leptin on tight junction-associated proteins in Sertoli cells and the possible leptin signaling pathways involved in this process. RESULTS: After treatment, there were no significant differences in body weights, reproductive organ weights, serum leptin levels and serum testosterone levels between leptin-treated mice and control mice. Administration of 3 mg/kg leptin reduced sperm concentration, motility and progressive motility while increasing the percentage of abnormal sperm and testicular cell apoptosis. Mice treated with 3 mg/kg leptin also had impaired blood testis barrier integrity, which was related to decreased tight junction-associated proteins in testes. Leptin directly reduced tight junction-associated proteins in Sertoli cells, JAK2/STAT, PI3K and ERK pathways were suggested to be involved in this process. CONCLUSIONS: Exogenous leptin negatively affects sperm parameters and impairs blood testis barrier integrity in mice. Leptin reduced tight junction-associated proteins in Sertoli cells, indicating that leptin has a direct role in impairing blood testis barrier integrity. Given the function of blood testis barrier in maintaining normal spermatogenesis, leptin-induced blood testis barrier impairment may be one of the mechanisms contributing to male subfertility and infertility.


Assuntos
Barreira Hematotesticular/efeitos dos fármacos , Barreira Hematotesticular/metabolismo , Leptina/farmacologia , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Animais , Relação Dose-Resposta a Droga , Leptina/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/metabolismo , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo
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