RESUMO
The gut microbiota is influenced by environmental factors such as food. Maternal diet during pregnancy modifies the gut microbiota composition and function, leading to the production of specific compounds that are transferred to the fetus and enhance the ontogeny and maturation of the immune system. Prebiotics are fermented by gut bacteria, leading to the release of short-chain fatty acids that can specifically interact with the immune system, inducing a switch toward tolerogenic populations and therefore conferring health benefits. In this study, pregnant BALB/cJRj mice were fed either a control diet or a diet enriched in prebiotics (Galacto-oligosaccharides/Inulin). We hypothesized that galacto-oligosaccharides/inulin supplementation during gestation could modify the maternal microbiota, favoring healthy immune imprinting in the fetus. Galacto-oligosaccharides/inulin supplementation during gestation increases the abundance of Bacteroidetes and decreases that of Firmicutes in the gut microbiota, leading to increased production of fecal acetate, which was found for the first time in amniotic fluid. Prebiotic supplementation increased the abundance of regulatory B and T cells in gestational tissues and in the fetus. Interestingly, these regulatory cells remained later in life. In conclusion, prebiotic supplementation during pregnancy leads to the transmission of specific microbial and immune factors from mother to child, allowing the establishment of tolerogenic immune imprinting in the fetus that may be beneficial for infant health outcomes.
Assuntos
Líquido Amniótico/metabolismo , Suplementos Nutricionais , Microbioma Gastrointestinal , Tolerância Imunológica , Prebióticos , Prenhez , Acetatos/metabolismo , Animais , Subpopulações de Linfócitos B/imunologia , Butiratos/metabolismo , Células Dendríticas/imunologia , Fezes/química , Fezes/microbiologia , Feminino , Feto/imunologia , Humanos , Inulina/administração & dosagem , Inulina/farmacologia , Troca Materno-Fetal , Camundongos , Camundongos Endogâmicos BALB C , Oligossacarídeos/administração & dosagem , Oligossacarídeos/farmacologia , Placenta/citologia , Placenta/imunologia , Gravidez , Resultado da Gravidez , Prenhez/imunologia , Prenhez/metabolismo , Efeitos Tardios da Exposição Pré-Natal , Propionatos/metabolismo , Ribotipagem , Subpopulações de Linfócitos T/imunologia , Útero/citologia , Útero/imunologiaRESUMO
Spontaneous abortion represents a common form of embryonic loss caused by early pregnancy failure. In the present study, we investigated the prophylactic effects of bee venom phospholipase A2 (bvPLA2), a regulatory T cell (Treg) inducer, on a lipopolysaccharide (LPS)-induced abortion mouse model. Fetal loss, including viable implants, the fetal resorption rate, and the fetal weight, were measured after LPS and bvPLA2 treatment. The levels of serum and tissue inflammatory cytokines were determined. To investigate the involvement of the Treg population in bvPLA2-mediated protection against fetal loss, the effect of Treg depletion was evaluated following bvPLA2 and LPS treatment. The results clearly revealed that bvPLA2 can prevent fetal loss accompanied by growth restriction in the remaining viable fetus. When the LPS-induced abortion mice were treated with bvPLA2, Treg cells were significantly increased compared with those in the non-pregnant, PBS, and LPS groups. After LPS injection, the levels of proinflammatory cytokines were markedly increased compared with those in the PBS mouse group, while bvPLA2 treatment showed significantly decreased TNF-α and IFN-γ expression compared with that in the LPS group. The protective effects of bvPLA2 treatment were not detected in Treg-depleted abortion-prone mice. These findings suggest that bvPLA2 has protective effects in the LPS-induced abortion mouse model by regulating Treg populations.
Assuntos
Aborto Espontâneo/tratamento farmacológico , Venenos de Abelha/enzimologia , Lipopolissacarídeos/toxicidade , Fosfolipases A2/uso terapêutico , Aborto Espontâneo/sangue , Aborto Espontâneo/induzido quimicamente , Aborto Espontâneo/imunologia , Animais , Citocinas/sangue , Citocinas/genética , Feminino , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Camundongos Endogâmicos C57BL , Fosfolipases A2/farmacologia , Gravidez , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia , Útero/efeitos dos fármacos , Útero/imunologiaRESUMO
During the transition from prepartum to early lactation, dairy cows often experience negative energy balance (NEB) that may result in reproductive stress and decreased fertility. The objective of this study was to observe the effects of rumen-protected methionine (RPM) on plasma amino acid concentrations, uterine cytology, immunohistochemistry (IHC) of glutathione peroxidase 1 (GPX) and superoxide dismutase 1 (SOD), and to confirm neutrophil extracellular trap (NET) formation. Multiparous Holstein cows (nâ¯=â¯20) were randomly assigned to two treatments starting at 21â¯d before calving until 73 days in milk (DIM). Treatments were: CON (nâ¯=â¯9, no supplementation, TMR with a Lys:Metâ¯=â¯3.5:1) and MET (n = 11, TMR + Smartamine® M with a Lys:Metâ¯=â¯2.8:1). Uterine endometrial biopsies, uterine cytology, and blood samples from the coccygeal artery or vein were collected at 15, 30, and 73 DIM. Blood plasma samples were analyzed for amino acids and metabolites. Uterine biopsies were analyzed for NET formation, neutrophil numbers, as well as GPX and SOD by IHC. Additionally, uterine cytology was analyzed for polymorphonuclear neutrophil (PMN) to epithelial cell percentage. Cows in CON had lower methionine plasma concentrations (18.05⯱â¯2.0⯵M) than cows in MET (30.39⯱â¯1.6⯵M). Cows in CON had greater cystine plasma concentrations (3.62⯱â¯0.3⯵M) than cows in MET (2.8⯱â¯0.3⯵M). No treatment differences were observed for SOD or GPX in the endometrium. Cows in CON tended to have a high score for positively immunolabeled GPX cells at 15 DIM than cows in MET. No treatment differences were observed for the percentage of PMN in uterine cytology, number of neutrophils, or extent of NET formation in the endometrium. A treatment by time interaction was observed for PMN percentage and the number of neutrophils: cows in MET tended to have greater PMN percentages than cows in CON at 15 DIM which decreased for subsequent days and cows in MET had greater neutrophil numbers in the endometrium at 30 DIM than cows in CON. In conclusion, dietary supplementation of RPM altered plasma amino acid concentrations and increased neutrophil infiltration in the postpartum period, suggesting improved uterine immunity.
Assuntos
Bovinos , Armadilhas Extracelulares/fisiologia , Metionina/farmacologia , Neutrófilos/fisiologia , Útero/fisiologia , Animais , Dieta/veterinária , Suplementos Nutricionais , Feminino , Lactação , Metionina/administração & dosagem , Rúmen , Útero/imunologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Bacterial endometritis is one of the major causes of reproductive disorders including infertility in farm animals. Antibiotics are generally used for treatment of such disorders but now a days residues of antibiotics are of great public health concern, therefore, phytoremediation is being considered as an alternative to use of antibiotics. AIM OF THE STUDY: Present study was undertaken to investigate the efficacy of Eucalyptus robusta leaves methanolic extract against endometritis along with the possible mechanism of action especially targeting inflammatory biomarkers. MATERIALS AND METHODS: Bacterial endometritis was produced using clinical isolates of E. coli and Staphyloccocus aureus from bovines (cows and buffaloes) endometritis cases. After seven days of inoculation of the mixed bacterial culture, endometritis was confirmed based on the presence of visible pus and edema, thinning of endometrial lining and presence of large number of polymorphonuclear cells and bacterial load in uterine flushing. Female Wistar rats were divided in to five groups namely control, sham-operated, endometritis, endometritis plus Eucalyptus leaves extract and endometritis plus cefixime. Serum specific inflammatory biomarkers (interleukin-1ß, interleukin-10, tumor necrosis factor-α, intercellular adhesion molecule-1, serum amyloid A) and myleoperoxidase, toll like receptors-4 and -9, inducible nitric oxide synthase, nitric oxide, cyclooxygenase 1 and 2 were estimated in uterine tissues using ELISA kits. RESULTS: Interleukin-10, serum amyloid A, myleoperoxidase, toll like receptors-4 and-9, cyclooxygenase-2, inducible nitric oxide synthase and nitric oxide were significantly increased while non significant increase in interleukin-1ß, cycloxygenase-1 and intercellular adhesion molecule-1 were observed but level of tumor necrosis factor-α was found decreased in rats of endometritis group. Histopathological lesions in uterus showed efficient induction of endometritis by presence of inflammatory cells which are lessened effectively after treatment with Eucalyptus leaves extract. Eucalyptus robusta leaves extract produced curative and protective effect against endometritis and results were comparable to or even better than cefixime. CONCLUSIONS: Eucalyptus robusta leaves extract possess promising antibacterial activity and efficacy against experimental endometritis and, therefore, holds promising potential for development of effective formulation for treatment of endometritis in animals.
Assuntos
Antibacterianos , Endometrite , Eucalyptus , Extratos Vegetais , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Ciclo-Oxigenase 2/metabolismo , Citocinas/sangue , Citocinas/imunologia , Endometrite/tratamento farmacológico , Endometrite/imunologia , Endometrite/metabolismo , Endometrite/patologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Feminino , Metanol/química , Óxido Nítrico Sintase Tipo II/metabolismo , Peroxidase/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Folhas de Planta , Ratos Wistar , Proteína Amiloide A Sérica/metabolismo , Solventes/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Receptor 4 Toll-Like/metabolismo , Receptor Toll-Like 9/metabolismo , Útero/efeitos dos fármacos , Útero/imunologia , Útero/metabolismo , Útero/patologiaRESUMO
Staphylococcus aureus (S. aureus) causes severe inflammation in various infectious diseases, leading to high mortality. The clinical application of antibiotics has gained a significant curative effect. However, it has led to the emergence of various resistant bacteria. Therefore, in this study, we investigated the protective effect of polydatin (PD), a traditional Chinese medicine extract, on S. aureus lipoteichoic acid (LTA)-induced injury in vitro and in vivo. First, a significant improvement in the pathological conditions of PD in vivo was observed, suggesting that PD had a certain protective effect on LTA-induced injury in a mouse model. To further explore the underlying mechanisms of this protective effect of PD, LTA-induced murine macrophages were used in this study. The results have shown that PD could reduce the NF-κB p65, and IκBα phosphorylation levels increased by LTA, resulting in a decrease in the transcription of pro-inflammatory factors, such as TNF-α, IL-1ß and IL-6. However, LTA can not only activate NF-κB through the recognition of TLR2 but also increase the level of intracellular reactive oxygen species (ROS), thereby activating NF-κB signalling. We also detected high levels of ROS that activate caspases 9 and 3 to induce apoptosis. In addition, using a specific NF-κB inhibitor that could attenuate apoptosis, namely NF-κB p65, acted as a pro-apoptotic transcription factor in LTA-induced murine macrophages. However, PD could inhibit the generation of ROS and NF-κB p65 activation, suggesting that PD suppressed LTA-induced injury by attenuating ROS generation and TLR2-NFκB signalling.
Assuntos
Antioxidantes/farmacologia , Endometriose/tratamento farmacológico , Glucosídeos/farmacologia , Substâncias Protetoras/farmacologia , Espécies Reativas de Oxigênio/antagonistas & inibidores , Estilbenos/farmacologia , Receptor 2 Toll-Like/genética , Fator de Transcrição RelA/genética , Animais , Sobrevivência Celular/efeitos dos fármacos , Endometriose/induzido quimicamente , Endometriose/genética , Endometriose/imunologia , Feminino , Regulação da Expressão Gênica , Proteínas I-kappa B/genética , Proteínas I-kappa B/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/isolamento & purificação , Lipopolissacarídeos/toxicidade , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Células RAW 264.7 , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Staphylococcus aureus/química , Ácidos Teicoicos/antagonistas & inibidores , Ácidos Teicoicos/isolamento & purificação , Ácidos Teicoicos/toxicidade , Receptor 2 Toll-Like/imunologia , Fator de Transcrição RelA/imunologia , Útero/efeitos dos fármacos , Útero/imunologia , Útero/patologiaRESUMO
During pregnancy, immune activity is tightly regulated so that antimicrobial protection of the mother and fetus is balanced with the need for immune tolerance to prevent fetal rejection. In this setting, the maternal-fetal interface, in the form of the uterine decidua, provides a heterogeneous immune cell population with the potential to mediate diverse activities throughout pregnancy. Recent studies have suggested that vitamin D may be a key regulator of immune function during pregnancy, with the fetal-maternal interface representing a prominent target. Among its non-classical actions are potent immunomodulatory effects, including induction of antibacterial responses and modulation of T-lymphocytes to suppress inflammation and promote tolerogenesis. Thus, vitamin D may play a pivotal role in normal decidual immune function by promoting innate responses to infection, while simultaneously preventing an over-elaboration of inflammatory adaptive immunity. Research to date has focused upon the potential role of vitamin D in preventing infectious diseases such as tuberculosis, as well as possibly suppressing of autoimmune disease. Nevertheless, vitamin D may also influence facets of immune function not immediately associated with primary innate responses. This review summarises our current understanding of decidual immune function with respect to the vitamin D metabolism and signalling, and as to how this may be affected by variations in maternal vitamin D status. There has recently been much interest in vitamin D supplementation of pregnant women, but our knowledge of how this may influence the function of decidua remains limited. Further insight into the immunomodulatory actions of vitamin D during pregnancy will help shed light upon this.
Assuntos
Troca Materno-Fetal/imunologia , Gravidez/imunologia , Vitamina D/fisiologia , Animais , Decídua/imunologia , Feminino , Humanos , Sistema Imunitário/fisiologia , Macrófagos/imunologia , Placenta/imunologia , Útero/citologia , Útero/imunologiaRESUMO
Selenium (Se) is an essential nutritional trace element that affects the development and function of the reproductive system. Endometritis is a reproductive obstacle disease that can seriously reduce the reproductive capacity of animal. To study the effects of dietary Se deficiency on lipopolysaccharide (LPS)-induced mice endometritis, we generated a model of LPS-induced mice endometritis. The Se content in uterine tissues was detected by fluorescence spectrophotometry. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß, and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR). The extent of phosphorylation of IκBα, NF-κB p65, ERK, JNK, and p38 and the expression of Toll-like receptor 4 (TLR4) were detected with Western blots. The TLR4 messenger RNA (mRNA) was analyzed with qRT-PCR. The results indicated that dietary Se intake significantly influenced Se levels in uterine tissues. The Se-deficient mice model was successfully replicated, and Se deficiency exacerbated uterine tissue histopathology; increased the expression of TNF-α, IL-1ß, and IL-6; facilitated the activation of TLR4; and enhanced the phosphorylation of IκBα, p65, ERK, JNK, and p38 in LPS-induced mice endometritis. Also, the effects were inhibited by a supplement of Se. In conclusion, our studies demonstrated that Se deficiency makes mice uterus more prone to inflammation. An appropriate Se supplement could enhance the immune condition of the uterus.
Assuntos
Endometrite/patologia , Inflamação/imunologia , Selênio/deficiência , Receptor 4 Toll-Like/metabolismo , Útero/patologia , Animais , Endometrite/induzido quimicamente , Ensaio de Imunoadsorção Enzimática , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Proteínas I-kappa B/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Lipopolissacarídeos , Camundongos , Camundongos Endogâmicos BALB C , Inibidor de NF-kappaB alfa , Fosforilação , RNA Mensageiro/genética , Distribuição Aleatória , Reação em Cadeia da Polimerase em Tempo Real , Selênio/farmacologia , Transdução de Sinais , Espectrometria de Fluorescência , Receptor 4 Toll-Like/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Útero/imunologia , Útero/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Uterine leiomyomas (fibroids, myomas) are the most common benign tumors of female reproductive tract. They are highly prevalent, with 70-80% of women burdened by the end of their reproductive years. Fibroids are a leading cause of pelvic pain, abnormal vaginal bleeding, pressure on the bladder, miscarriage, and infertility. They are the leading indication for hysterectomy, and costs exceed 6 billion dollars annually in the United States. Unfortunately, no long-term medical treatments are available. Dysregulation of inflammatory processes are thought to be involved in the initiation of leiomyoma and extracellular matrix deposition, cell proliferation, and angiogenesis are the key cellular events implicated in leiomyoma growth. In modern pharmaceutical industries, dietary phytochemicals are used as source of new potential drugs for many kinds of tumors. Dietary phytochemicals may exert therapeutic effects by interfering with key cellular events of the tumorigenesis process. At present, a negligible number of phytochemicals have been tested as therapeutic agents against fibroids. In this context, our aim was to introduce some of the potential dietary phytochemicals that have shown anti-inflammatory, antiproliferative, antifibrotic, and antiangiogenic activities in different biological systems. This review could be useful to stimulate the evaluation of these phytochemicals as possible therapies for uterine fibroids.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Suplementos Nutricionais , Leiomioma/prevenção & controle , Compostos Fitoquímicos/uso terapêutico , Útero/imunologia , Animais , Proliferação de Células , Feminino , Fibrose , Humanos , Leiomioma/dietoterapia , Leiomioma/imunologia , Leiomioma/fisiopatologia , Neovascularização Patológica/etiologia , Neovascularização Patológica/prevenção & controle , Neovascularização Fisiológica , Útero/irrigação sanguínea , Útero/citologia , Útero/patologiaRESUMO
Toll like receptors (TLRs) are pattern-recognition molecules that initiate the innate immune response to pathogens. Pulmonary surfactant protein (SP)-A is an endogenously produced ligand for TLR2 and TLR4. SP-A has been proposed as a fetally produced signal for the onset of parturition in the mouse. We examined the effect of interactions between SP-A and the pathogenic TLR agonists lipopolysaccharide (LPS), peptidoglycan (PGN) and polyinosinic:cytidylic acid (poly(I:C)) (ligands for TLR4, TLR2 and TLR3, respectively) on the expression of inflammatory mediators and preterm delivery. Three types of mouse macrophages (the cell line RAW 264.7, and fresh amniotic fluid and peritoneal macrophages, including macrophages from TLR4 and TLR2 knockout mice) were treated for up to 7 hours with pathogenic TLR agonists with or without SP-A. SP-A alone had no effect upon inflammatory mediators in mouse macrophages and did not independently induce preterm labor. SP-A significantly suppressed TLR ligand-induced expression of inflammatory mediators (interleukin (IL)-1ß, tumor necrosis factor (TNF)-α and the chemokine CCL5) via a TLR2 dependent mechanism. In a mouse inflammation-induced preterm delivery model, intrauterine administration of SP-A significantly inhibited preterm delivery, suppressed the expression of proinflammatory mediators and enhanced the expression of the CXCL1 and anti-inflammatory mediator IL-10. We conclude that SP-A acts via TLR2 to suppress TLR ligand-induced preterm delivery and inflammatory responses.
Assuntos
Anti-Inflamatórios/uso terapêutico , Nascimento Prematuro/prevenção & controle , Proteína A Associada a Surfactante Pulmonar/uso terapêutico , Receptor 2 Toll-Like/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Feminino , Feto/efeitos dos fármacos , Feto/imunologia , Feto/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Placenta/efeitos dos fármacos , Placenta/imunologia , Placenta/metabolismo , Gravidez , Nascimento Prematuro/imunologia , Proteína A Associada a Surfactante Pulmonar/farmacologia , Receptor 2 Toll-Like/agonistas , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Útero/efeitos dos fármacos , Útero/imunologia , Útero/metabolismoRESUMO
This study was conducted to explore the abortifacient effect and the mechanisms of the Chinese herbal medicine component toosendanin, and to elucidate the significance of the Th1 cytokines IFN-gamma and TNF-alpha, CD4+ and CD8+ T lymphocytes in the occurrence of abortion. Graded doses of toosendanin were given by intraperitoneal injection (i.p.) to mice at day 5, 6, 7 of gestation. The levels of Th1 cytokines (IFN-gamma, TNF-alpha) in serum and uterine tissues from mice sacrificed at day 8 were analyzed by enzyme linked immunosorbent assay (ELISA). Presence of T lymphocytes in endometrium was detected by immunohistochemistry. The results revealed that injection of toosendanin could produce a dose-dependent toxicity. The IFN-gamma, TNF-alpha content in serum and uterine tissues were increased significantly. The CD4+ and CD8+ T lymphocytes were also increased in the endometrium of toosendanin treated groups. In conclusion, toosendanin is pregnancy-toxic to animals and it is relevant to the increased contents of IFN-gamma, TNF-alpha and CD4+, CD8+ T lymphocytes.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Medicamentos de Ervas Chinesas/farmacologia , Interferon gama/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Útero/efeitos dos fármacos , Animais , Ensaio de Imunoadsorção Enzimática , Feminino , Interferon gama/sangue , Troca Materno-Fetal , Camundongos , Camundongos Endogâmicos BALB C , Gravidez , Fator de Necrose Tumoral alfa/sangue , Útero/imunologiaRESUMO
Supplementation of semen extender with caffeine and CaCl(2) for artificial insemination (AI) of fresh spermatozoa has been demonstrated to reduce recruitment of uterine polymorphonuclear leukocytes (PMNs) and the activity of phagocytosis. Here, we determined if addition of caffeine and CaCl(2) to semen extender improves the fertility of frozen-thawed boar semen. In experiment 1, gilts were cervically inseminated twice with frozen-thawed boar spermatozoa (25 x 10(8) cells per dose) suspended in Modena solution (n=7) or modified Beltsville Thawing Solution supplemented with caffeine and CaCl(2) (BCC, n=7). The gilts were slaughtered 4 h later, and their oviducts and uterine horns plus the body of the uterus were flushed to recover PMNs and non-phagocytosed spermatozoa. There was no difference in the total number of uterine PMNs between gilts inseminated with Modena solution and those inseminated with BCC (3.8 x 10(8) vs. 1.5 x 10(8) cells, respectively); however, the total number of uterine spermatozoa was higher when gilts were inseminated with BCC (40.6 x 10(6) cells) compared with those inseminated with Modena solution (1.4 x 10(6) cells, P<0.05). In experiment 2, gilts and sows were subjected to intrauterine insemination twice with frozen-thawed spermatozoa suspended (25 x 10(8) sperm per dose) in Modena (n=21) or BCC (n=21). The overall pregnancy and farrowing rates were higher in females inseminated with BCC (71.4 and 61.9%, respectively) compared with those inseminated with Modena solution (38.1 and 28.6%, respectively, P<0.05). However, no significant difference in litter size of piglets was observed between treatments (7.2 +/- 1.6 piglets for Modena solution vs. 8.2 +/- 0.9 piglets for BCC solution). In conclusion, we demonstrated that use of BCC solution for frozen-thawed boar semen produced better pregnancy and farrowing rates following AI than Modena solution, probably by reducing the phagocytosis of spermatozoa.
Assuntos
Cafeína/farmacologia , Cloreto de Cálcio/farmacologia , Criopreservação/veterinária , Crioprotetores/farmacologia , Inseminação Artificial/veterinária , Preservação do Sêmen/veterinária , Sus scrofa/fisiologia , Criação de Animais Domésticos/métodos , Criação de Animais Domésticos/estatística & dados numéricos , Animais , Sobrevivência Celular/efeitos dos fármacos , Criopreservação/métodos , Sinergismo Farmacológico , Feminino , Fertilização/efeitos dos fármacos , Inseminação Artificial/métodos , Nascido Vivo , Masculino , Oviductos/citologia , Oviductos/efeitos dos fármacos , Gravidez , Taxa de Gravidez , Preservação do Sêmen/métodos , Transporte Espermático/efeitos dos fármacos , Útero/citologia , Útero/efeitos dos fármacos , Útero/imunologiaRESUMO
The aim of the present work was to study some of the adverse effects produced by hyperandrogenism on the uterine function. Daily injection of dehydroepiandrosterone (DHEA: 6 mg/ 100 g body weight, sc) for 20 consecutive days induced polycystic ovaries in BALB/c mice. In this model, we found that DHEA produced alterations on uterine histology closely related to the development of tumour structures. In addition, hyperandrogenism induced a pro-inflammatory and a pro-oxidant condition represented by increased levels of prostaglandin F2 alpha production and uterine nitric oxide synthase (NOS) activity and by a decrease in both superoxide dismutase (SOD) and catalase (CAT) activities together with a decrease in the levels of the antioxidant metabolite glutathione (GSH). DHEA also induced an increase in CD4+ together with a decrease in the CD8+ T lymphocytes that infiltrate the uterine tissue. We conclude that this intricate network of regulators could be responsible for the low rate of implantation observed in women with polycystic ovary syndrome.
Assuntos
Adjuvantes Imunológicos/toxicidade , Desidroepiandrosterona/toxicidade , Hiperandrogenismo/fisiopatologia , Síndrome do Ovário Policístico/induzido quimicamente , Útero/fisiopatologia , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Catalase/antagonistas & inibidores , Catalase/metabolismo , Ciclo-Oxigenase 1/efeitos dos fármacos , Ciclo-Oxigenase 2/efeitos dos fármacos , Dinoprosta/biossíntese , Feminino , Glutationa/antagonistas & inibidores , Glutationa/metabolismo , Hiperandrogenismo/patologia , Proteínas de Membrana/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico Sintase/efeitos dos fármacos , Óxido Nítrico Sintase/metabolismo , Ovário/patologia , Estresse Oxidativo/efeitos dos fármacos , Síndrome do Ovário Policístico/imunologia , Síndrome do Ovário Policístico/patologia , Prostaglandinas E/biossíntese , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/metabolismo , Útero/imunologia , Útero/patologiaRESUMO
BACKGROUND: Since the two estrogen receptor isoforms ERalpha and ERbeta have been discovered it is unclear by which receptor immunomodulating or feminizing effects are mediated. In this study, the effects of the two selective ERalpha- and ERbeta-agonists ethinylestradiol and biochaninA, respectively, on acute cardiac allograft rejection, uterus growth, vascular adhesion molecule and MHC-II expression were investigated and verified using in vitro cell culture. METHODS: Heterotopic Lewis to ovarectomized F344 cardiac transplantations were performed. The study groups received supplemental biochaninA or ethinylestradiol, the control group received no treatment. Grafts and uteri were harvested on the fifth postoperative day and blood was taken for hormone plasma level quantifications. Purified Lewis aortic endothelial cell cultures were pre-treated with biochaninA or ethinylestradiol and stimulated with TNF-alpha or IFN-gamma for quantification of ICAM-1/VCAM-1 and MHC-II expression. Endothelium-lymphocyte adhesion assays were performed using purified F344 lymphocytes. RESULTS: Both biochaninA and ethinylestradiol treatment significantly reduced graft mononuclear infiltration of CD8(+) and ED1(+) cells and markedly reduced ISHLT grading compared to untreated controls. Either agent significantly inhibited lymphocyte adhesion, endothelial VCAM-1 upregulation during graft rejection and during TNF-alpha-stimulation in vitro, whereas no effect was observed for ICAM-1 upregulation. BiochaninA but not ethinylestradiol significantly reduced endothelial MHC-II upregulation in vivo and in vitro. Only ethinylestradiol treatment strongly affected uterus growth in ovarectomized recipients. CONCLUSIONS: Only the treatment with the phytoestrogen biochaninA reduced endothelial MHC-II expression in vivo and in vitro and weakened allograft rejection without affecting the reproductive system. Supplemental phytoestrogens may therefore provide further benefits in the clinical setting.
Assuntos
Genisteína/uso terapêutico , Rejeição de Enxerto/prevenção & controle , Transplante de Coração/imunologia , Fitoestrógenos/uso terapêutico , Doença Aguda , Animais , Adesão Celular , Moléculas de Adesão Celular/metabolismo , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Feminino , Genisteína/sangue , Genisteína/farmacologia , Antígenos de Histocompatibilidade Classe II/imunologia , Linfócitos/imunologia , Tamanho do Órgão/efeitos dos fármacos , Fitoestrógenos/sangue , Fitoestrógenos/farmacologia , Ratos , Ratos Endogâmicos Lew , Receptores de Estrogênio/agonistas , Receptores de Estrogênio/análise , Útero/anatomia & histologia , Útero/efeitos dos fármacos , Útero/imunologiaRESUMO
OBJECTIVE: To explore the effect of Jiantaiye (JTY) on the expression of estrogen receptor (ER) and ER mRNA in uterus of mice with embryo implantation dysfunction. METHODS: Embryo implantation dysfunction mouse models were induced with mifepristone and treated with JTY. All animals were sacrificed on day 8 of pregnancy. The endometrial ER and ER mRNA expressions were assessed by immunnohistochemical SP method and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Area ratio and absorbency of ER in the JTY treated group's gland and stroma were higher than those of the model group, quite similar to those of the normal control's, and ER mRNA expression in treated group's uterus was significantly higher than that in the models, but it was not significantly different from the normal control. CONCLUSION: JTY improves the endometrial development by increasing ER and ER mRNA expressions of uterus of mice with embryo implantation dysfunction.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Implantação do Embrião , Receptores de Estrogênio/metabolismo , Doenças Uterinas/fisiopatologia , Útero/efeitos dos fármacos , Útero/imunologia , Animais , Endométrio/efeitos dos fármacos , Endométrio/metabolismo , Feminino , Imuno-Histoquímica/métodos , Masculino , Camundongos , Camundongos Endogâmicos , Gravidez , RNA Mensageiro/metabolismo , Receptores de Estrogênio/genética , Coloração e Rotulagem , Distribuição Tecidual , Doenças Uterinas/imunologiaRESUMO
In this study the ability of the phytoestrogen genistein (GEN) to regulate proliferation in the rat uterine tissue and the associated molecular mechanisms were investigated in a dose and time dependent manner. A single administration of GEN induced a rapid increase of the uterine weight during the first 24 h. In contrast to E2, treatment with GEN for 3 days did not result in a further increase of the uterine weight. GEN only marginally effected the thickness of the uterine epithelium and the expression of epithelial proliferating cell nuclear antigen (PCNA). Whereas, estrogen sensitive genes were modulated significantly, the expression of key genes involved in the regulation of proliferation (PCNA, ERalpha /ERbeta ratio) remained unaffected by GEN. Our results indicate that GEN has only a limited ability to activate molecular mechanisms involved in the induction of proliferation whereas estrogen sensitive genes are induced in a estrogen like manner.
Assuntos
Estradiol/farmacologia , Expressão Gênica/efeitos dos fármacos , Genisteína/farmacologia , Fitoestrógenos/farmacologia , Útero/efeitos dos fármacos , Animais , Proliferação de Células/efeitos dos fármacos , Epitélio/efeitos dos fármacos , Epitélio/imunologia , Feminino , Antígeno Nuclear de Célula em Proliferação/análise , Antígeno Nuclear de Célula em Proliferação/genética , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores de Esteroides/análise , Receptores de Esteroides/genética , Útero/citologia , Útero/imunologiaRESUMO
The role of mast cells and their main secretory products in the effects of oestradiol on the uterus was investigated. Ovariectomized rats were treated with a single injection of oestradiol (10 micrograms per rat, i.m.) or vehicle together with drugs affecting the activity of mast cells, cromoglycate (10 mg per rat, i.m.), which diminishes the degranulation of mast cells, or compound 48/80 (0.5 mg per rat, i.m.), which enhances this process. Oestradiol or vehicle was also administered with two important secretory products of mast cells, heparin (0.4 mg per rat, i.m.) or histamine (2 mg per rat, i.m.). All drugs were injected simultaneously with oestradiol (first injection) and then every 6 h until the animals were killed. Observations were performed at 24, 36 and 48 h after oestradiol or vehicle injection. The condition of mast cells was determined by the percentage of degranulated mast cells in sections stained with toluidine blue. Oestradiol-induced effects in the uterus were estimated by the mitotic index, proliferating cell nuclear antigen-labelling index, DNA content, volumes of cells, nuclei and nucleoli in the luminal epithelium, glandular epithelium and stroma cells of the endometrium. Cromoglycate treatment resulted in a decrease in both mast cell degranulation and all examined oestradiol effects in the uterus at all periods of observation. Compound 48/80 increased mast cell degranulation and expression of one aspect of oestradiol effects on the volumes of cell compartments. Histamine or heparin led to a marked increase in the cell, nucleus and nucleolus volumes in all uterine structures. However, heparin produced a depression in proliferation, whereas histamine had a weak transient stimulating action on this process. No effects of the protocols were found in the absence of oestradiol treatment. These results suggest that mast cells are involved in the realization of oestrogen action, including the stimulation of cell growth and proliferation in the uterus, and that the effect of mast cells is mediated by both histamine and heparin.
Assuntos
Degranulação Celular/efeitos dos fármacos , Estradiol/farmacologia , Mastócitos/fisiologia , Útero/efeitos dos fármacos , Útero/imunologia , Análise de Variância , Animais , Cromolina Sódica/farmacologia , Feminino , Heparina/farmacologia , Histamina/farmacologia , Mastócitos/efeitos dos fármacos , Índice Mitótico/efeitos dos fármacos , Azeite de Oliva , Ovariectomia , Óleos de Plantas/farmacologia , Ratos , Útero/citologia , p-Metoxi-N-metilfenetilamina/farmacologiaRESUMO
Pseudopregnancy, induced by mating females with vasectomized males, is a frequently used model for studying pregnancy-like uterine changes in the absence of an embryo. Leukocytes make a significant contribution to uterine cellularity during pregnancy. The present study was designed to determine whether changes in numbers and distribution of leukocytes in the uterus during pseudopregnancy and following intraluminal injection of a deciduogenic stimulus parallel changes observed during the first eight days of pregnancy. Common leukocyte antigen positive (CLA+) cells, macrophages (F4/80+ cells), and granulocytes were assessed between days 1 and 8 of pseudopregnancy using qualitative and quantitative immunohistochemistry. High numbers of CLA+ leukocytes were present on days 1 and 2. Those were comprised primarily by macrophages, polymorphonuclear leukocytes, and eosinophils. High concentrations of leukocytes were detected in the endometrium, and some granulocytes were observed migrating through the luminal epithelium. Leukocytes, principally macrophages, were reduced in number and were distributed throughout the endometrium on days 3 and 4. Introduction of oil to the uterine lumen on day 4 stimulated primary decidualization. Decidual cells were CLA- and F4/80-, and, as decidualization proceeded, CLA+ and F4/80+ cells decreased in number in the anti-mesometrial uterus and were detected primarily in the deep endometrium. Later, a secondary decidualization zone developed in the mesometrial aspect of the uterus. Unlike the initial decidual reaction, which was relatively free of leukocytes, the secondary decidual zone contained very high numbers of CLA+ and F4/80+ cells. The uninjected uterine horn remained relatively unchanged from days 3 through 8.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Leucócitos/patologia , Pseudogravidez/patologia , Útero/patologia , Animais , Antígenos de Superfície/análise , Biomarcadores , Quimiotaxia de Leucócito , Feminino , Inflamação/induzido quimicamente , Contagem de Leucócitos , Macrófagos/patologia , Masculino , Camundongos , Pseudogravidez/imunologia , Óleo de Gergelim/toxicidade , Útero/imunologiaRESUMO
The efficiency of usage of the preparations of thymalin in hyperplasia of endometrium has been studied in the experiment on 125 female guinea pigs. Indicated pathology was caused by the introduction of oil solution of synestrol during 19 weeks. It is determined that marked hyperplasia of endometrium is developing as a result of prolonged introduction of synestrol in experimental animals and the decrease of the amount of T- and B-lymphocytes takes place. The use of thymalin results in normalization of indexes of immune system and in partial normalization of morphologic structure of endometrium.