RESUMEN
Leydig cells (Lc), located in the interstitial space of the testis between seminiferous tubules, produce 95% of testosterone in male individuals, which is pivotal for male sexual differentiation, spermatogenesis, and maintenance of the male secondary sex characteristics. Lc are prone to senescence in aging testes, resulting in compromised androgen synthesis capability upon aging. However, little is known about whether Lc undergo senescence in a chronic inflammatory environment. To investigate this question, mouse models of experimental autoimmune orchitis (EAO) were used, and Lc were analyzed by high throughput scRNA-Seq. Data were screened and analyzed by correlating signaling pathways with senescence, apoptosis, androgen synthesis, and cytokine/chemokine signaling pathways. EAO did induce Lc senescence, and Lc senescence in turn antagonized androgen synthesis. Based on the correlation screening of pathways inducing Lc senescence, a plethora of pathways were found to play potential roles in triggering Lc senescence during EAO, among which the Arf6 and angiopoietin receptor pathways were highly correlated with senescence signature. Notably, complement and interstitial fibrosis activated by EAO worsened Lc senescence and strongly antagonized androgen synthesis. Furthermore, most proinflammatory cytokines enhanced both senescence and apoptosis in Lc and spermatogonia (Sg) during EAO, and proinflammatory cytokine antagonism of the glutathione metabolism pathway may be key in inducing cellular senescence during EAO.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Senescencia Celular/inmunología , Activación de Complemento/inmunología , Fibrosis/inmunología , Células Intersticiales del Testículo/inmunología , Angiopoyetinas/inmunología , Animales , Apoptosis/genética , Apoptosis/inmunología , Enfermedades Autoinmunes/genética , Senescencia Celular/genética , Activación de Complemento/genética , Citocinas/inmunología , Modelos Animales de Enfermedad , Fibrosis/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Transducción de Señal/genética , Transducción de Señal/inmunología , Análisis de la Célula Individual/métodosRESUMEN
This study aimed to analyze the effect and mechanism of immunization of oral KISS1 DNA vaccine on the proliferation of goat testicular Leydig cells. Ten 8-week-old male goats were randomly divided into KISS1 DNA vaccine and control groups for immunization (five goats each group). These goats were sacrificed at 8 weeks after primary immunization, and the tissue samples of hypothalamus, pituitary, and testis and Leydig cell samples were collected for RT-PCR and CCK8 assay. Immunization with the oral KISS1 DNA vaccine effectively inhibited the proliferation of Leydig cells, the expression of hypothalamus KISS1, GPR54, and GnRH mRNA, pituitary GnRHR and LH mRNA, testicular LHR mRNA, and apoptosis-inhibitory gene Bcl-2 mRNA in Leydig cells. By contrast, the immunization enhanced the mRNA expression of apoptosis-promoting gene Bax and Clusterin in Leydig cells. These findings indicate that immunization with the oral KISS1 DNA vaccine can inhibit the proliferation of goat testicular Leydig cells mainly via the hypothalamic-pituitary-testicular axis and apoptosis-related genes.
Asunto(s)
Proliferación Celular , Anticonceptivos Masculinos , Cabras , Kisspeptinas , Células Intersticiales del Testículo , Vacunas de ADN , Animales , Masculino , Anticoncepción Inmunológica/veterinaria , Regulación de la Expresión Génica/inmunología , Kisspeptinas/inmunología , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/fisiología , Receptores de Kisspeptina-1/genética , Receptores de Kisspeptina-1/metabolismo , Receptores de HL/genética , Receptores de HL/metabolismo , Receptores LHRH/genética , Receptores LHRH/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Testosterona/metabolismo , Vacunas de ADN/inmunologíaRESUMEN
The immune privilege of the testes is necessary to prevent immune attacks to gamete-specific antigens and paternal major histocompatibility complex (MHC) antigens, allowing for normal spermatogenesis. However, infection and inflammation of the male genital tract can break the immune tolerance and represent a significant cause of male infertility. Different T cell subsets have been identified in mammalian testes, which may be involved in the maintenance of immune tolerance and pathogenic immune responses in testicular infection and inflammation. We reviewed the evidence in the published literature on different T subtypes (regulatory T cells, helper T cells, cytotoxic T cells, γδ T cells, and natural killer T cells) in human and animal testes that support their regulatory roles in infertility and the orchitis pathology. While many in vitro studies have indicated the regulation potential of functional T cell subsets and their possible interaction with Sertoli cells, Leydig cells, and spermatogenesis, both under physiological and pathological processes, there have been no in situ studies to date. Nevertheless, the normal distribution and function of T cell subsets are essential for the immune privilege of the testes and intact spermatogenesis, and T cell-mediated immune response drives testicular inflammation. The distinct function of different T cell subsets in testicular homeostasis and the orchitis pathology suggests a considerable potential of targeting specific T cell subsets for therapies targeting chronic orchitis and immune infertility.
Asunto(s)
Inmunidad , Linfocitos T/inmunología , Linfocitos T/metabolismo , Testículo/inmunología , Testículo/metabolismo , Animales , Autoinmunidad , Biomarcadores , Manejo de la Enfermedad , Susceptibilidad a Enfermedades , Homeostasis , Humanos , Inmunomodulación , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/metabolismo , Masculino , Células de Sertoli/inmunología , Células de Sertoli/metabolismo , Espermatogénesis , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismoRESUMEN
Interleukin (IL)-18 is an inflammasome-mediated cytokine produced by germ cells, Leydig cells, and resident macrophages that is indispensable in the maintenance of homeostasis in the testis. We previously demonstrated that endogenous IL-18 induces testicular germ cell apoptosis during acute inflammation when plasma IL-18 levels are very high. However, the impact of acute inflammation and IL-18 on Leydig cells remained unclear. TM3 cells, a mouse Leydig cell line, and RAW264.7 cells, a mouse macrophage cell line, were stimulated with lipopolysaccharide (LPS) or recombinant IL-18 (rIL-18). We assessed the expression of inflammatory cytokines, caspase cleavage, and markers of apoptotic pathways. In Leydig cells, caspase 3 cleavage was increased and death-receptor-mediated apoptotic pathways were activated after LPS stimulation. However, LPS stimulation did not increase IL-18 expression in the Leydig cell line. When high-dose rIL-18 was administered to the Leydig cell line to mimic levels seem after inflammation, rIL-18 upregulated Tnf-α mRNA, Fadd mRNA, and Fas protein, promoted cleavage of caspase-8 and caspase-3, and induced apoptosis. Low-dose rIL-18 did not stimulate apoptosis. To determine if the high level of IL-18 seen in the testes after inflammation was derived from immune cells, we examined IL-18 protein expression in a macrophage cell line, RAW264.7. In contrast to the TM3 cells, IL-18 was significantly increased in RAW264.7 cells after LPS stimulation. These results suggest that high-dose IL-18 derived from macrophages is harmful to Leydig cells. Reducing the overexpression of IL-18 could be a new therapeutic approach to prevent Leydig cell apoptosis as a result of acute inflammation.
Asunto(s)
Apoptosis/inmunología , Enfermedad Crítica , Interleucina-18/metabolismo , Células Intersticiales del Testículo/patología , Orquitis/inmunología , Animales , Modelos Animales de Enfermedad , Proteína de Dominio de Muerte Asociada a Fas/metabolismo , Humanos , Inflamasomas/metabolismo , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/metabolismo , Lipopolisacáridos/inmunología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Orquitis/patología , Células RAW 264.7 , Transducción de Señal/inmunologíaRESUMEN
Long-term use of cell phones emitting electromagnetic fields (EMFs) have raised concerns regarding public health in recent year. We aimed to investigate the possible effects of 900â¯MHz EMF exposure (60â¯min/day for 28 days) on the rat testis. Another objective was to determine whether the deleterious effect of EMF radiation would be reduced by the administration of thymoquinone (TQ) (10â¯mg/kg/day). Twenty-four male adult Wistar albino rats were randomly selected, then assigned into four groups as followControl, EMF, TQ and EMFâ¯+â¯TQ. Testicular samples were analyzed using histological, stereological, biochemical and immunohistochemical techniques. Total numbers of primary spermatocytes and spermatids as well as Leydig cells were significantly decreased in the EMF group compared to the Control group (pâ¯<â¯0.05). In the EMFâ¯+â¯TQ group, the total number of primary spermatocytes was significantly increased compared to the EMF group (pâ¯<â¯0.05). Superoxide dismutase (SOD) activity was significantly increased in the EMF group compared to the Control group (p < 0.05). Also, serum testosterone levels and wet weight of testes were significantly decreased in the EMF group compared to the Control group (pâ¯<â¯0.05). Our findings suggested that exposure to a 900â¯MHz EMF had adverse effects on rat testicular tissue and that the administration of TQ partially mitigated testicular oxidative damages caused by EMF radiation.
Asunto(s)
Benzoquinonas/farmacología , Teléfono Celular , Células Intersticiales del Testículo , Ondas de Radio/efectos adversos , Espermátides , Espermatocitos , Animales , Inmunohistoquímica , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/metabolismo , Células Intersticiales del Testículo/patología , Masculino , Ratas , Ratas Wistar , Espermátides/inmunología , Espermátides/metabolismo , Espermátides/patología , Espermatocitos/inmunología , Espermatocitos/metabolismo , Espermatocitos/patologíaRESUMEN
Infections of the reproductive tract are known to contribute to testicular inflammatory impairment, leading to an increase of pro-inflammatory cytokines such as IL-1ß, and a decline in sperm quality. Prokineticin 2 (PK2), a secretory protein, is closely associated with the secretion of pro-inflammatory cytokines in inflamed tissue. It was reported that increased PK2 is related to the upregulation of IL-1ß, but the underlying mechanism remains elusive. Here, we illustrated that PK2 was upregulated in testicular macrophages (TM) in a rat model of uropathogenic Escherichia coli (UPEC) infection, which induced the activation of the NLRP3 inflammasome pathway to boost IL-1ß secretion. Administration of PK2 inhibitor alleviated the inflammatory damage and suppressed IL-1ß secretion. Moreover, PK2 promoted NLRP3 expression and the release of cleaved IL-1ß from TM to the supernatants after the challenge with UPEC in vitro. IL-1ß in the supernatants affected Leydig cells by suppressing the expression of genes encoding for the enzymes P450scc and P450c17, which are involved in testosterone production. Overall, we revealed that increased PK2 levels in TM in UPEC-induced orchitis may impair testosterone synthesis via the activation of the NLRP3 pathway. Our study provides a new insight into the mechanisms underlying inflammation-associated male infertility and suggests an anti-inflammatory therapeutic target for male infertility.
Asunto(s)
Hormonas Gastrointestinales/inmunología , Inflamasomas/inmunología , Macrófagos/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Neuropéptidos/inmunología , Orquitis/inmunología , Testículo/inmunología , Escherichia coli Uropatógena/inmunología , Animales , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/microbiología , Hormonas Gastrointestinales/genética , Hormonas Gastrointestinales/metabolismo , Expresión Génica/inmunología , Humanos , Inflamasomas/metabolismo , Inflamación/genética , Inflamación/inmunología , Inflamación/metabolismo , Interleucina-1beta/inmunología , Interleucina-1beta/metabolismo , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/metabolismo , Macrófagos/metabolismo , Masculino , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Neuropéptidos/genética , Neuropéptidos/metabolismo , Orquitis/microbiología , Ratas Wistar , Transducción de Señal/inmunología , Esteroide 17-alfa-Hidroxilasa/genética , Esteroide 17-alfa-Hidroxilasa/inmunología , Esteroide 17-alfa-Hidroxilasa/metabolismo , Testículo/metabolismo , Infecciones Urinarias/inmunología , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/fisiologíaRESUMEN
It was reported previously that adult mouse stem Leydig cells (SLCs) express CD51 (integrin α-chain V). However, it is still unclear whether all CD51+â¯cells are SLCs. In the present study, we found that CD51+â¯cells can be classified into two sub-groups, a weakly-staining group (CD51+) and a strongly-staining group (CD51++). The CD51+ cells expressed common SLC marker genes, including Nestin, Pdgfra and Coup-tf2, while CD51++ cells did not express these genes. Instead, they expressed macrophage markers, such as F4/80, Cd115 and Tnfa. When these cells were induced to differentiate in vitro, the CD51+ cells, but not CD51++ cells, formed Leydig cells. Overall, our results showed that although SLCs expressed CD51, not all CD51-expressing cells are SLCs. The cells that expressed high levels of CD51 are actually macrophages.
Asunto(s)
Integrina alfaV/metabolismo , Células Intersticiales del Testículo/citología , Células Madre/inmunología , Testículo/citología , Animales , Biomarcadores/metabolismo , Diferenciación Celular , Células Cultivadas , Células Intersticiales del Testículo/inmunología , Macrófagos/citología , Macrófagos/inmunología , Masculino , Ratones , Espermatogénesis , Células Madre/citología , Testículo/inmunología , Regulación hacia ArribaRESUMEN
The mammalian testis possesses a special immunological environment because of its properties of remarkable immune privilege and effective local innate immunity. The testicular immune privilege protects immunogenic germ cells from systemic immune attack, and local innate immunity is important in preventing testicular microbial infections. Thus, this study aimed to immunohistochemically demonstrate the distribution and localization of CD68-, CD8-, MHCI- and MHCII-positive immune cells in the testes and epididymes. Negative immunoreactivity was detected in the seminiferous tubule epithelium and peritubular myoid cells of the testes upon staining in CD68, CD8 and MHC Class I. Positive CD68 immunoreaction was determined in the Sertoli cells and some Leydig cells. The detection of positive cells for CD8 clearly indicated the presence of lymphocytes. Furthermore, the staining with MHCI intensity was ascertained to vary from weak to moderate in the Sertoli and Leydig cells and connective tissue cells. MHCII-positive immunoreactivity was determined in myoid cells and Leydig cells in the interstitial area. The epithelium of the epididymis showed positive staining for CD68 and CD8, but the stroma displayed a rather weak staining. In the ram epididymis, neither intraepithelial nor interstitial positive reaction was observed for MHCI. In the epididymis, the basal cells displayed a stronger staining for MHCII. In conclusion, these cells not only contribute to local immunity through their direct effects on the quality of fertility in males, but also contribute either directly or indirectly to immune privilege by minimizing the development of both autoimmune reactions and potentially harmful risks.
Asunto(s)
Bovinos/anatomía & histología , Epidídimo/citología , Ovinos/anatomía & histología , Testículo/citología , Animales , Antígenos CD/análisis , Antígenos CD/inmunología , Antígenos de Diferenciación Mielomonocítica/análisis , Antígenos de Diferenciación Mielomonocítica/inmunología , Antígenos CD8/análisis , Antígenos CD8/inmunología , Bovinos/inmunología , Epidídimo/inmunología , Genes MHC Clase I/inmunología , Genes MHC Clase II/inmunología , Inmunidad Innata , Inmunohistoquímica/veterinaria , Células Intersticiales del Testículo/inmunología , Masculino , Células de Sertoli/inmunología , Ovinos/inmunología , Testículo/inmunologíaRESUMEN
Mumps virus (MuV) infection frequently causes orchitis and impairs male fertility. However, the mechanisms underlying the innate immune responses to MuV infection in the testis have yet to be investigated. This study showed that MuV induced innate immune responses in mouse Sertoli and Leydig cells through TLR2 and retinoic acid-inducible gene I (RIG-I) signaling, which result in the production of proinflammatory cytokines and chemokines, including TNF-α, IL-6, MCP-1, CXCL10, and type 1 interferons (IFN-α and IFN-ß). By contrast, MuV did not induce the cytokine production in male germ cells. In response to MuV infection, Sertoli cells produced higher levels of proinflammatory cytokines and chemokines but lower levels of type 1 IFNs than Leydig cells did. The MuV-induced cytokine production by Sertoli and Leydig cells was significantly reduced by the knockout of TLR2 or the knockdown of RIG-I signaling. The local injection of MuV into the testis triggered the testicular innate immune responses in vivo. Moreover, MuV infection suppressed testosterone synthesis by Leydig cells. This is the first study examining the innate immune responses to MuV infection in testicular cells. The results provide novel insights into the mechanisms underlying the MuV-induced innate immune responses in the testis.
Asunto(s)
Inmunidad Innata , Células Intersticiales del Testículo/inmunología , Virus de la Parotiditis/inmunología , Paperas/inmunología , Paperas/virología , Células de Sertoli/inmunología , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Factor 3 Regulador del Interferón/metabolismo , Células Intersticiales del Testículo/metabolismo , Células Intersticiales del Testículo/virología , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Ratones Noqueados , Paperas/genética , Paperas/metabolismo , FN-kappa B/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Receptores de Superficie Celular , Células de Sertoli/metabolismo , Células de Sertoli/virología , Testosterona/biosíntesis , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismoRESUMEN
BACKGROUND: The chronic nature of tuberculosis and the protracted immuno-inflammatory reactions are implied in a series of metabolic and immune-endocrine changes accompanying the disease. We explored components from the hypothalamous-pituitary-gonadal axis and their relationship with cytokines involved in disease immunopathology, in male TB patients. METHODS: Plasma samples from 36 active untreated pulmonary TB male patients were used to determine TNF-α, IFN-γ, TGF-ß, IL-6, cortisol, dehydroepiandrosterone, testosterone, progesterone, estradiol, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by ELISA. Healthy controls corresponded to 21 volunteers without contact with TB patients and similar age (40 ± 16,8 years). Testicular histological samples from necropsies of patients dying from TB were immune-stained for IL-1ß, TNF-α, IL-6 and IFN-γ. The TM3 mouse Leydig cell line was incubated with recombinants TNF-α, IFN-γ and TGF-ß, supernatants were collected and used to measure testosterone by ELISA. RESULTS: Patients showed decreased levels of testosterone in presence of high amounts of LH, together with augmented IFN-γ, IL-6 and TGF-ß levels. Testicular histological sections showed abundant presence of IL-1ß, TNF-α, IL-6 and IFN-γ in interstitial macrophages, Sertoli cells and some spermatogonia. In vitro treatment of Leydig cells with these cytokines led to a remarkable reduction of testosterone production.
Asunto(s)
Andrógenos/sangre , Citocinas/sangre , Mediadores de Inflamación/sangre , Testículo/metabolismo , Testosterona/sangre , Tuberculosis Pulmonar/sangre , Adulto , Animales , Estudios de Casos y Controles , Línea Celular , Citocinas/farmacología , Humanos , Mediadores de Inflamación/farmacología , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Testículo/efectos de los fármacos , Testículo/inmunología , Testículo/patología , Tuberculosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/patologíaRESUMEN
BACKGROUND: Although the testis is considered an immunoprivileged organ it can orchestrate immune responses against pathological insults such as infection and trauma. Experimental autoimmune orchitis (EAO) is a model of chronic inflammation whose main histopathological features it shares with human orchitis. In EAO an increased number of macrophages infiltrate the interstitium concomitantly with progressive germ cell degeneration and impaired steroidogenesis. Up-regulation of nitric oxide (NO)-NO synthase (NOS) system occurs, macrophages being the main producers of NO. OBJECTIVE: The aim of our study was to evaluate the role of NO-NOS system in orchitis development and determine the involvement of NO released by testicular macrophages on germ cell apoptosis and testosterone secretion. METHOD AND RESULTS: EAO was induced in rats by immunization with testicular homogenate and adjuvants (E group) and a group of untreated normal rats (N) was also studied. Blockage of NOS by i.p. injection of E rats with a competitive inhibitor of NOS, L-NAME (8mg/kg), significantly reduced the incidence and severity of orchitis and lowered testicular nitrite content. L-NAME reduced germ cell apoptosis and restored intratesticular testosterone levels, without variations in serum LH. Co-culture of N testicular fragments with testicular macrophages obtained from EAO rats significantly increased germ cell apoptosis and testosterone secretion, whereas addition of L-NAME lowered both effects and reduced nitrite content. Incubation of testicular fragments from N rats with a NO donor DETA-NOnoate (DETA-NO) induced germ cell apoptosis through external and internal apoptotic pathways, an effect prevented by N-acetyl-L-cysteine (NAC). DETA-NO inhibited testosterone released from Leydig cells, whereas NAC (from 2.5 to 15 mM) did not prevent this effect. CONCLUSIONS: We demonstrated that NO-NOS system is involved in the impairment of testicular function in orchitis. NO secreted mainly by testicular macrophages could promote oxidative stress inducing ST damage and interfering in Leydig cell function.
Asunto(s)
Enfermedades Autoinmunes/prevención & control , Inhibidores Enzimáticos/farmacología , Células Intersticiales del Testículo/metabolismo , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/antagonistas & inhibidores , Orquitis/prevención & control , Espermatozoides/metabolismo , Acetilcisteína/farmacología , Adyuvantes Inmunológicos , Animales , Apoptosis/efectos de los fármacos , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Técnicas de Cocultivo , Mezclas Complejas , Regulación de la Expresión Génica , Humanos , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/inmunología , Células Intersticiales del Testículo/patología , Hormona Luteinizante/sangre , Hormona Luteinizante/genética , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/patología , Masculino , Óxido Nítrico/biosíntesis , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Orquitis/inducido químicamente , Orquitis/inmunología , Orquitis/patología , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Espermatozoides/efectos de los fármacos , Espermatozoides/inmunología , Espermatozoides/patología , Testosterona/biosíntesis , Testosterona/metabolismo , Triazenos/farmacologíaRESUMEN
PROBLEM: Previous studies demonstrated a strong association between low androgen levels and reduced capacity to mount an inflammatory response. However, the mechanisms underlying these observations are largely not understood. METHODS OF STUDY: Generation of CD4+CD25+Foxp3+ regulatory T cells in Leydig cell-conditioned media was determined by flow cytometry and ELISA. Influence of testosterone on cytokine response was measured in LPS-stimulated testicular macrophages, Sertoli and peritubular cells. RESULTS: Leydig cell-conditioned media dose-dependently stimulated expression of transcription factor Foxp3 and secretion of IL-10 in splenic CD4+ T cells, an effect abolished by addition of the anti-androgen flutamide. In isolated Sertoli and peritubular cells, testosterone pre-treatment suppressed the LPS-induced inflammatory response on TNF-α mRNA expression, while no effect was evident in testicular macrophages (TM). CONCLUSIONS: Androgens can influence the immune system under normal conditions by the generation and functional differentiation of regulatory T cells and in testicular inflammation by direct effect on Sertoli and peritubular cells.
Asunto(s)
Factores de Transcripción Forkhead/biosíntesis , Interleucina-10/biosíntesis , Células Intersticiales del Testículo/inmunología , Linfocitos T Reguladores/citología , Testosterona/metabolismo , Antagonistas de Andrógenos/farmacología , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/inmunología , Células Cultivadas , Quimiocina CCL2/biosíntesis , Medios de Cultivo Condicionados/farmacología , Flutamida/farmacología , Inflamación/inmunología , Interleucina-10/metabolismo , Macrófagos/inmunología , Masculino , ARN Mensajero/biosíntesis , Ratas , Ratas Wistar , Células de Sertoli/inmunología , Linfocitos T Reguladores/inmunología , Testosterona/antagonistas & inhibidores , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Adiponectin is an adipocyte hormone that is predominantly secreted by adipocytes, and has important roles in glucose and lipid homeostasis. Recent studies have shown that adiponectin is also involved in the regulation of many endocrine organs, such as the ovary, adrenal gland, and pituitary. However, its biological role in male testes is largely unexplored. The present findings demonstrate the presence of adeponectin receptors (adiponectin receptor 1 and adiponectin receptor 2) in TM3 cells derived from mouse Leydig cells. Proinflammatory cytokine treatment significantly downregulated mRNA and protein levels of adiponectin receptor 1 and adiponectin receptor 2. However, adiponectin pretreatment successfully inhibited the signaling pathway mediated by proinflammatory cytokines. At the molecular level, we provide compelling evidence that adeponectin achieves this by suppressing nuclear factor-κB activation through promotion of AMP-activated protein kinase phosphorylation. Thus, our data clearly indicate that adiponectin plays a protective role in Leydig cells through its anti-inflammatory actions.
Asunto(s)
Adiponectina/metabolismo , Citocinas/antagonistas & inhibidores , Regulación hacia Abajo , Células Intersticiales del Testículo/metabolismo , FN-kappa B/antagonistas & inhibidores , Receptores de Adiponectina/metabolismo , Transducción de Señal , 3-Hidroxiesteroide Deshidrogenasas/antagonistas & inhibidores , 3-Hidroxiesteroide Deshidrogenasas/química , 3-Hidroxiesteroide Deshidrogenasas/genética , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Proteínas Quinasas Activadas por AMP/metabolismo , Animales , Línea Celular , Ciclooxigenasa 2/química , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Silenciador del Gen , Células Intersticiales del Testículo/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/química , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosfoproteínas/agonistas , Fosfoproteínas/antagonistas & inhibidores , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Fosforilación , Procesamiento Proteico-Postraduccional , Receptores de Adiponectina/antagonistas & inhibidores , Receptores de Adiponectina/genética , Regulación hacia ArribaRESUMEN
Late-onset hypogonadism (LOH) is closely related to secondary androgen deficiency in aged males, but the mechanism remains unclear. In this study, we found that reduced testosterone production in aged rat Leydig cells is associated with decreased autophagic activity. Primary rat Leydig cells and the TM3 mouse Leydig cell line were used to study the effect of autophagic deficiency on Leydig cell testosterone production. In Leydig cells from young and aged rats, treatment with wortmannin, an autophagy inhibitor, inhibited luteinising hormone (LH)-stimulated steroidogenic acute regulatory (StAR) protein expression and decreased testosterone production. In contrast, treatment with rapamycin, an autophagy activator, enhanced LH-stimulated steroidogenesis in Leydig cells from aged, but not young, rats. Intracellular reactive oxygen species (ROS) levels were increased in both young and aged Leydig cells treated with wortmannin but decreased only in aged Leydig cells treated with rapamycin. Furthermore, an increased level of ROS, induced by H(2)O(2), resulted in LH-stimulated steroidogenic inhibition. Finally, knockdown of Beclin 1 decreased LH-stimulated StAR expression and testosterone production in TM3 mouse Leydig cells, which were associated with increased intracellular ROS level. These results suggested that autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells, which might be influenced by intracellular ROS levels.
Asunto(s)
Autofagia , Células Intersticiales del Testículo/inmunología , Animales , Proteínas Reguladoras de la Apoptosis/genética , Beclina-1 , Western Blotting , Células Cultivadas , Técnicas de Silenciamiento del Gen , Células Intersticiales del Testículo/citología , Células Intersticiales del Testículo/metabolismo , Hormona Luteinizante/fisiología , Masculino , Ratones , Microscopía Electrónica de Transmisión , Fosfoproteínas/metabolismo , Radioinmunoensayo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Testosterona/biosíntesis , Testosterona/metabolismoRESUMEN
The testis is an immunoprivileged site, where the local cell-initiated testicular innate immune responses play a crucial role in defense against microbial infections. Mechanisms modulating the testicular cell-built defense system remain to be clarified. In this article, we demonstrate that Leydig cells, a major cell population in the testicular interstitium, initiate innate immunity through the activation of Toll-like receptors (TLRs). Several TLRs are expressed in mouse Leydig cells; among these, TLR3 and TLR4 are expressed at relatively high levels compared with other TLR members. Both TLR3 and TLR4 can be activated by their agonists (polyinosinic:polycytidylic acid and lipopolysaccharide) in Leydig cells and subsequently induce the production of inflammatory factors, such as IL-1ß, IL-6, TNF-α, and type 1 interferons (IFN) (IFN-α and IFN-ß). Notably, the activation of TLR3 and TLR4 suppresses steroidogenesis by Leydig cells. Further, we provide evidence that Axl and Mer receptor tyrosine kinases are expressed in Leydig cells and regulate TLR-mediated innate immune responses negatively. Data presented here describe a novel function of Leydig cells in eliciting testicular innate immune responses that should contribute to the protection of the testis from microbial infections.
Asunto(s)
Inmunidad Innata , Células Intersticiales del Testículo/inmunología , Receptores Toll-Like/metabolismo , Animales , Células Cultivadas , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Citocinas/genética , Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Células Intersticiales del Testículo/citología , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Fosfoproteínas/metabolismo , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , ARN Mensajero/metabolismo , Proteínas Tirosina Quinasas Receptoras/genética , Proteínas Tirosina Quinasas Receptoras/metabolismo , Esteroide 17-alfa-Hidroxilasa/metabolismo , Testículo/citología , Testículo/inmunología , Testículo/metabolismo , Receptor Toll-Like 3/agonistas , Receptor Toll-Like 3/antagonistas & inhibidores , Receptor Toll-Like 3/genética , Receptor Toll-Like 3/metabolismo , Receptor Toll-Like 4/agonistas , Receptor Toll-Like 4/antagonistas & inhibidores , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Receptores Toll-Like/agonistas , Receptores Toll-Like/antagonistas & inhibidores , Receptores Toll-Like/genética , Tirosina Quinasa c-Mer , Tirosina Quinasa del Receptor AxlRESUMEN
PROBLEM: This study examined the effect of lipopolysaccharide (LPS) on the capacity of Leydig cells to produce and express interleukin-18 (IL-18), IL-18 receptor (IL-18R) and the IL-1beta-converting enzyme (ICE) (IL-18 family), under in vitro conditions. METHOD OF STUDY: Primary Leydig cells (LCs) were isolated from murine testis by the Percoll technique, and cultured both in the presence and absence of LPS (0.1, 1, 5 microg/mL) for 3 and 24 hr. LCs were examined for their capacity to produce and express IL-18 family molecules by using immunohistochemical staining (IHC), enzyme-linked immunosorbent assay (ELISA), Western blot and real-time polymerase chain reaction (PCR) analysis. RESULTS: Leydig cells were shown to constitutively express IL-18, as examined by IHC, ELISA, Western blot and real-time PCR analysis. Addition of LPS to LC cultures was shown to significantly increase the basal levels of IL-18, in a dose- and time course-dependent manner, as examined by ELISA, Western blot and real-time PCR analysis. In addition, LPS increased LC cultures to express ICE and IL-18 R, as examined by real-time PCR analysis. CONCLUSION: Our results demonstrate that LPS increased the capacity of murine LCs to produce the IL-18 family molecules. IL-18, in the testis, might be involved in the regulation of physiological and infection/inflammatory processes, and thus, could be a component of the autocrine/paracrine factor net that controls steroidogenesis and male fertility; further studies are needed to confirm this possibility.
Asunto(s)
Caspasa 1/metabolismo , Interleucina-18/metabolismo , Células Intersticiales del Testículo/metabolismo , Lipopolisacáridos/inmunología , Receptores de Interleucina-18/metabolismo , Animales , Células Cultivadas , Células Intersticiales del Testículo/citología , Células Intersticiales del Testículo/inmunología , Masculino , RatonesRESUMEN
BACKGROUND: The immuno-privileged status of the testis is essential to the maintenance of its functions, and innate immunity is likely to play a key role in limiting harmful viral infections, as demonstrated in the rat. In men mumps virus infects Leydig cells and has deleterious effects on testosterone production and spermatogenesis. The aim of this study was to test whether mumps virus infection of isolated human Leydig cells was associated with an inhibition of their innate antiviral defences. METHODS: Leydig cell production of mRNA and protein for interferons (IFNs) and of three antiviral proteins-2'5' oligoadenylate synthetase (2'5'OAS), double-stranded RNA-activated protein kinase (PKR) and MxA-was investigated, in the absence or presence of mumps virus or viral stimuli including poly I:C, a mimetic of RNA viruses replication product. RESULTS: Stimulated or not, human Leydig cells appeared unable to produce routinely detectable IFNs alpha, beta and gamma. Although the level of PKR remained unchanged after stimulation, the expression of 2'5'OAS and MxA was enhanced following either mumps virus or poly I:C exposure (P < 0.05 versus control). CONCLUSIONS: Overall, our results demonstrate that mumps virus replication in human Leydig cells is not associated with a specific inhibition of IFNs or 2'5'OAS, MxA and PKR production and that these cells display relatively weak endogenous antiviral abilities, as opposed to their rat counterparts.
Asunto(s)
Antivirales/farmacología , Inmunidad Innata/fisiología , Inductores de Interferón/farmacología , Células Intersticiales del Testículo/virología , Virus de la Parotiditis/inmunología , Poli I-C/farmacología , 2',5'-Oligoadenilato Sintetasa/metabolismo , Células Cultivadas , Proteínas de Unión al GTP/metabolismo , Humanos , Interferones/metabolismo , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/inmunología , Masculino , Virus de la Parotiditis/patogenicidad , Proteínas de Resistencia a Mixovirus , Replicación Viral/inmunología , eIF-2 Quinasa/metabolismoRESUMEN
AIM: To study the regulation of immune function of rat Leydig cells by vasoactive intestinal peptide (VIP). METHODS: Rat Leydig cells were seperated and infected by UU, with or without the incubation with VIP. The expression of FasL on Leydig cells in different group was analysed by flow cytometry(FCM). The mRNA expression of IL-1, IL-6, TGF-beta and FasL of Leydig cells in different group was identified by RT-PCR. SD rats were infected by UU with or without the injection of VIP, and the testis tissnes of each group were har vested and observed under transmission electron microscope. RESULTS: When testis was infected by UU, VIP up-regulated the mRNA expression of IL-1, IL-6, and TGF-beta, and down-regulated the expression of FasL in vitro. In addition, there was significant difference in testis morphous of rats from different group. CONCLUSION: VIP could regulate the expression pattern of IL-1, IL-6, TGF-beta and FasL by Leydig cells, which may contribute to maintain immune privilege of the testis. VIP could regulate rat Leydig cell immune function.
Asunto(s)
Fármacos Gastrointestinales/farmacología , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/inmunología , Péptido Intestinal Vasoactivo/farmacología , Animales , Células Cultivadas , Proteína Ligando Fas/genética , Proteína Ligando Fas/metabolismo , Interleucina-1/genética , Interleucina-6/genética , Células Intersticiales del Testículo/metabolismo , Células Intersticiales del Testículo/microbiología , Masculino , Microscopía Electrónica de Transmisión , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Testículo/citología , Testículo/efectos de los fármacos , Testículo/metabolismo , Testículo/ultraestructura , Factor de Crecimiento Transformador beta/genética , Ureaplasma urealyticum/inmunología , Ureaplasma urealyticum/patogenicidadRESUMEN
The present study in the wall lizard, Hemidactylus flaviviridis, was aimed to understand the role of histamine (HA) in the regulation of Leydig cell and testicular macrophage activities, for the first time, in ectothermic vertebrates. Although HA did not affect the testosterone production from unstimulated Leydig cells, it had dual concentration-related effects, stimulatory at a low concentration of 10(-10) M while inhibitory at a high concentration of 10(-5) M, on FSH-induced testosterone production. This suggests that HA did not influence the basal Leydig cell steroidogenesis, but modulated the FSH-stimulated testosterone production in a biphasic manner depending upon its concentration. However, HA failed to affect the FSH-stimulated Leydig cell proliferation, indicating that HA modulated the testosterone production from Leydig cells without influencing their proliferation in wall lizards. HA, apart from Leydig cells, differentially regulated the testicular macrophage immune responses. It inhibited phagocytosis and superoxide production at high concentration (10(-5) M), while stimulated superoxide production and could not affect phagocytosis at low concentration (10(-10) M). Using selective H1 and H2 antagonists, pyrilamine and famotidine respectively, H1 receptor subtype was seen responsible for mediating the inhibitory effect of HA on Leydig cell steroidogenesis and testicular macrophage immune responses at high concentration, while H2 receptors were involved for the stimulation at low concentration.
Asunto(s)
Histamina/farmacología , Células Intersticiales del Testículo/inmunología , Lagartos , Macrófagos/fisiología , Testosterona/biosíntesis , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Famotidina/farmacología , Antagonistas de los Receptores Histamínicos/farmacología , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/metabolismo , Macrófagos/efectos de los fármacos , Masculino , Fagocitosis/efectos de los fármacos , Pirilamina/farmacología , Receptores Histamínicos H1/metabolismo , Receptores Histamínicos H2/metabolismo , Superóxidos/metabolismoRESUMEN
The tubuli recti (TR) are immunologically special, because the lymphocytes preferably accumulate around them during the course of T-cell dependent testicular autoimmunity in mice. This finding implies that the testicular interstitium around the TR is where autoreactive lymphocytes can gain access to autoimmunogenic germ cell antigens. In the present study, the histoarchitecture of the TR was minutely examined in normal mice. Three-dimensional analysis showed that 14-16 TR appeared to be connected to the rete testis (RT). Electron microscopical analysis revealed that the epithelial cells in the TR formed protruding cytoplasmic strings, with active endocytosis of degenerated spermatozoa, and exhibited three different morphological characteristics. Furthermore, a few macrophages were found to have penetrated into the TR. Immunohistochemical image analysis revealed that more macrophages specifically accumulated around the TR than the RT or seminiferous tubules. These findings indicate that macrophages preferentially accumulate around the TR, where contact between germ cell antigens and macrophages may occur under normal and pathological conditions.