RESUMEN
Mucositis is an inflammation of the gastrointestinal mucosa resulting from high doses of radio/chemotherapy treatment and may lead to interruption of antineoplasic therapy. Soluble fibres, like pectin, increase SCFA production, which play a role in gut homoeostasis and inflammation suppression. Due to the properties of pectin, the aim of the present study was to evaluate the effect of a high-fibre (HF) diet on chemotherapy-induced mucositis in a murine model. C57/BL6 mice received control (AIN93M), HF, low/zero fibre (LF) diets for 10 d prior to mucositis challenging with irinotecan (75 mg/kg), or they were treated with acetate added to drinking water 5 d prior to and during the mucositis induction. Mice that received the HF diet showed decreased immune cells influx and improved histopathological parameters in the intestine, compared with mice that received the normal diet. Furthermore, the HF diet decreased intestinal permeability induced in the mucositis model when compared with the control group. This effect was not observed for acetate alone, which did not improve gut permeability. For instance, mice that received the LF diet had worsened gut permeability, compared with mice that received the normal diet and mucositis. The effects of the HF and LF diets were shown to modulate the intestinal microbiota, in which the LF diet increased the levels of Enterobacteriaceae, a group associated with gut inflammation, whereas the HF diet decreased this group and increased Lactobacillus and Bifidobacterium (SCFA producers) levels. In conclusion, the results demonstrated the importance of dietary fibre intake in the modulation of gut microbiota composition and homoeostasis maintenance during mucositis in this model.
Asunto(s)
Antineoplásicos , Fibras de la Dieta/administración & dosificación , Mucositis , Animales , Antineoplásicos/efectos adversos , Modelos Animales de Enfermedad , Inflamación , Ratones , Mucositis/inducido químicamente , PectinasRESUMEN
Fermented whey dairy beverages are dairy products obtained by fermentation from a mixture of milk and whey. These beverages have important health benefits, which could be improved with the addition of probiotic cultures. This study assessed the protective effect of the cosupplementation of a probiotic culture (Lactobacillus casei 01) with a fermented whey dairy beverage against infection by Salmonella enterica ssp. enterica serovar Typhimurium in a murine model. Two fermented whey dairy beverages were prepared: conventional (FWB; starter culture) and probiotic (PFWB; starter and probiotic cultures). In the first set of experiments, Balb/C female mice were treated with FWB or PFWB, challenged with Salmonella Typhimurium, and analyzed for clinical signs, weight loss, and mortality for 20 d postinfection. In the second set of experiments, mice were treated with FWB or PFWB, challenged with Salmonella Typhimurium, and killed on d 10 postinfection. The liver, colon, and ileum were used for myeloperoxidase, eosinophil peroxidase, and histological analysis and translocation to the liver. The contents from the small intestine were used for secretory IgA determination. The FWB treatment showed a better effect on animal survival (70%), translocation of the pathogen to the liver (2 out of 10), histopathology (fewer lesions), and inflammation than PFWB, which presented 50% animal survival, translocation in 5 out of 10 animals, and higher lesions. The control group presented 40% animal survival, translocation in 6 out of 10 animals, and severe lesions. Therefore, FWB was deemed to have a greater protective effect against Salmonella Typhimurium infection in the murine model compared with PFWB.
Asunto(s)
Productos Lácteos Cultivados , Salmonelosis Animal/prevención & control , Salmonella typhimurium , Suero Lácteo , Animales , Bebidas , Femenino , Promoción de la Salud , Inmunoglobulina A Secretora/análisis , Inflamación/prevención & control , Intestino Delgado/inmunología , Intestino Delgado/patología , Lacticaseibacillus casei/fisiología , Hígado/microbiología , Hígado/patología , Ratones , Ratones Endogámicos BALB C , Probióticos , Salmonelosis Animal/inmunología , Salmonelosis Animal/patología , Proteína de Suero de LecheRESUMEN
A healthy skin provides a protective barrier against pathogenic micro-organisms. Recent studies have shown that probiotics, as those of Bifidobacterium genus, could act beneficially in dermatology, both when ingested and by topical use. In the present study, we evaluated by in vitro antagonism assays and using two skin cell lines the potential of four strains of Bifidobacterium spp. Among the four bifidobacteria, Bifidobacterium longum 51A was the only one able to inhibit the growth of the eight pathogenic indicators tested. Production of some cytokines and extracellular matrix proteins was determined when ccc or inactivated cells of the bifidobacteria were incubated with keratinocyte and/or fibroblast cell cultures. Significant results were observed only for IL-6, IL-8 and IL-18 production, and inactivated Bifidobacterium pseudolongum 1191A was the only one which significantly stimulated collagen production, whereas lumican was stimulated by treatments with live Bifidobacterium bifidum 1622A , B. longum 51A and B. pseudolongum 1191A . Highest adhesion and internalization capabilities were observed with B. bifidum 1622A and Bifidobacterium breve 1101A . Concluding, B. longum 51A was highlighted for its antagonistic capacity and B. bifidum 1622A and B. pseudolongum 1191A for stimulating the production of cytokines and proteins of the extracellular matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: The skin is the first line of defence against invasive micro-organisms, and its local microbiota provides additional protective functions based on antagonism against pathogenic micro-organisms and immunomodulation. Based on in vitro assays using Bifidobacterium spp. we demonstrated the antagonistic potential, as well as capacity in stimulating the production of cytokines and proteins of the extracellular matrix that these bacteria may exert on skin cells. This positive influence suggests the use of a consortium of these bifidobacteria in a topical product for dermatological treatments.
Asunto(s)
Antibiosis/fisiología , Bifidobacterium/metabolismo , Citocinas/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Probióticos/metabolismo , Piel/microbiología , Bifidobacterium/clasificación , Candida albicans/crecimiento & desarrollo , Línea Celular , Humanos , Malassezia/crecimiento & desarrollo , Propionibacterium acnes/crecimiento & desarrollo , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus epidermidis/crecimiento & desarrolloRESUMEN
This study evaluated the expression of FSH receptors (FSHR) in the different stages of goat follicle development and investigated whether the addition of increasing concentrations of FSH throughout the culture period influences the survival, growth and antral formation of in vitro-cultured caprine preantral follicles. The expression of FSHR was analysed before and after culturing follicles using real-time RT-PCR. For the culture, preantral follicles (≥150 µm) were isolated from ovarian fragments and cultured for 18 days in α-MEM+ alone or associated with recombinant FSH (rFSH: 100 or 1000 ng/ml), or in α-MEM+ supplemented with increasing concentrations of FSH throughout culture periods as follows: (a) sequential medium 1: FSH 100 ng/ml (from day 0 to 6), FSH 500 ng/ml (from day 6 to 12) and FSH 1000 ng/ml (from day 12 to 18); and (b) sequential medium 2: FSH 500 ng/ml (from day 0 to 9) and 1000 ng/ml (from day 9 to 18). Follicle development was evaluated on the basis of antral cavity formation, follicular and oocyte growth, and cumulus-oocyte complex health. The expression of FSHR in isolated caprine follicles increased from the preantral to antral phase. Regarding the culture, after 18 days, sequential medium 1 promoted follicular survival, antrum formation and a reduction in oocyte extrusion. Both sequential media promoted a higher rate of meiotic resumption compared with the other treatments. In conclusion, the addition of increased concentrations of FSH (sequential medium) has a significant impact on the in vitro development of caprine preantral follicles.
Asunto(s)
Folículo Ovárico/citología , Folículo Ovárico/fisiología , Receptores de HFE/genética , Animales , Células Cultivadas , Medios de Cultivo , Femenino , Hormona Folículo Estimulante/farmacología , Cabras , Hormonas/farmacología , Técnicas In Vitro , Oocitos/citología , Oocitos/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , ARN Mensajero/genética , Receptores de HFE/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Allergic contact dermatitis (ACD) is a common allergic skin disease that affects individuals subjected to different antigen exposure conditions and significantly impacts the quality of life of those affected. Numerous studies have demonstrated that probiotics suppress inflammation through immunomodulatory effects. In this study, we aimed to evaluate the effect of the probiotic Bifidobacterium longum 51A as a preventive treatment for ACD using an oxazolone-induced murine model. We demonstrated that B. longum 51A exerted a prophylactic effect on oxazolone-induced ACD-like skin inflammation via reductions in ear and dermal thickness and leucocyte infiltration. The administration of inactivated B. longum 51A did not affect oxazolone-induced ACD-like skin inflammation, suggesting that the bacteria must be alive to be effective. Given that B. longum 51A is an acetate producer, we treated mice with acetate intraperitoneally, which also prevented ear and dermal thickening. Moreover, the tissue levels of the inflammatory cytokines and chemokines interleukin (IL)-10, IL-33, tumour necrosis factor-α, chemokine (C-C motif) ligand 2/monocyte chemoattractant protein-1 and chemokine (C-C motif) ligand 5/RANTES were significantly reduced after probiotic treatment, but only IL-33 and IL-10 were reduced when the mice were treated with acetate. These results show that B. longum 51A exerted a potential prophylactic effect on skin inflammation and that acetate represents one potential mechanism. However, other factors are likely involved since these two treatments do not yield the same results.
Asunto(s)
Bifidobacterium longum/fisiología , Dermatitis Alérgica por Contacto/inmunología , Dermatitis Alérgica por Contacto/prevención & control , Probióticos/administración & dosificación , Animales , Citocinas/genética , Citocinas/inmunología , Dermatitis Alérgica por Contacto/etiología , Dermatitis Alérgica por Contacto/genética , Femenino , Humanos , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-33/genética , Interleucina-33/inmunología , Ratones , Ratones Endogámicos BALB C , Oxazolona/efectos adversos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The aim of this study was to investigate the effects of estradiol and follicle-stimulating hormone (FSH) on survival and growth of caprine preantral follicles. Pieces of ovarian tissue were cultured for 1 or 7 days in minimum essential medium (MEM) containing estradiol (1, 5, 10, 20 or 40 pg/ml), FSH (50 ng/ml), or a combination of the two hormones. Cultured and noncultured control ovarian tissues were processed for histological and ultrastructural studies. The results showed that after 7 days of culture, the treatments that yielded the highest percentage of normal follicles relative to MEM alone were those that combined FSH with estradiol at 1, 5 or 20 pg/ml. The addition of FSH to 1-day cultures containing 1 pg/ml estradiol or to 7-day cultures with 1 or 5 pg/ml estradiol increased the percentage of normal follicles compared to estradiol alone at the same concentrations. After 7 days of culture, all treatments generated higher percentages of developing follicles as compared to control and MEM alone. The addition of either FSH or 10 pg/ml of estradiol to the culture media or estradiol (1, 5, 10 or 20 pg/ml) and FSH in combination significantly increased follicular diameter as compared with MEM alone following 7 days of culture. Ultrastructural studies confirmed follicular integrity after 7 days of culture in the presence of 1 pg/ml estradiol plus FSH. In conclusion, this study demonstrated that the interaction between estradiol and FSH maintains ultrastructural integrity and stimulates activation and further growth of cultured caprine preantral follicles.
Asunto(s)
Estradiol/administración & dosificación , Hormona Folículo Estimulante/administración & dosificación , Hormonas/administración & dosificación , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/crecimiento & desarrollo , Animales , Técnicas de Cultivo de Célula , Supervivencia Celular , Medios de Cultivo/química , Relación Dosis-Respuesta a Droga , Estradiol/química , Femenino , Hormona Folículo Estimulante/química , Cabras , Hormonas/química , Microscopía Electrónica de Transmisión , Folículo Ovárico/ultraestructura , Factores de TiempoRESUMEN
The aim of this study was to evaluate the effect of vasoactive intestinal peptide (VIP)on the survival, activation and growth of goat preantral follicles after in vitro culture. The ovarian cortex was divided into small pieces and one fragment was immediately fixed (control). The remaining fragments were cultured in vitro for 1 or 7 days at 39 degrees C and 5% CO(2), in supplemented minimum essential medium (MEM(+)) with or without different concentrations of VIP (1, 10, 50, 100 or 200 ng/ml). Noncultured (fresh control) and cultured ovarian fragments were processed for histological analysis and transmission electron microscopy. Follicles were classified as primordial or developing, and as normal or degenerated. Our findings indicate that when compared with control, addition of all concentrations of VIP except 200 ng/ml resulted in similar percentages of normal preantral follicles after 1 and 7 days of culture. Culture of ovarian cortex tissue for 1 and 7 days increased the percentage of follicular activation in all treatments when compared with control, except with 1 ng/ml of VIP after 1 day. However, no difference was observed between VIP-treated and MEM(+)-treated follicles. In addition, after 7 days of culture, the highest follicular and oocyte diameters were observed in follicles cultured with 10 ng/ml VIP relative to MEM(+) alone. Transmission electron microscopy showed ultrastructural integrity of follicles after 7 days of culture in 10 ng/ml VIP. In conclusion, this study demonstrates that VIP maintains follicular integrity and stimulates caprine preantral follicle growth.
Asunto(s)
Folículo Ovárico/fisiología , Péptido Intestinal Vasoactivo/farmacología , Animales , Medios de Cultivo/farmacología , Femenino , Cabras , Técnicas de Cultivo de Tejidos , Péptido Intestinal Vasoactivo/administración & dosificaciónRESUMEN
Food allergy is triggered when there is an abnormal activation of the immune system by food allergens. Currently, there is no curative therapy for this pathological condition. Due to the immunomodulatory properties of probiotics they are potential candidates as therapeutic tools for food allergy. Therefore, the aim of this study was to evaluate the probiotic effect of Saccharomyces cerevisiae UFMG A-905 (905) in an in vivo model of food allergy. Probiotic effect was assessed by clinical, histological, immunological and microbiological parameters analysis. Furthermore, we also evaluated if 905 after inactivation has an effect, as well as if such an effect is dose dependent. Our results showed that oral administration of only viable 905 promotes a significant attenuation of tissue injury and myeloperoxidase (MPO) activity levels. Moreover, the treatment reduced interleukin 17 levels, and administration of the supernatant from the yeast culture also promoted a significant decrease in MPO levels. However, considering the systemic parameters, immunoglobulin (Ig)E and IgG anti-ovalbumin, which are essentials for triggering the allergic process, there was no effect, suggesting that the yeast promotes a local but not a systemic effect in the model evaluated. In addition, we found that only high doses of viable 905 were able to attenuate the signs of inflammation. In conclusion, oral administration of 905 led to a local effect that depends on the viability of the yeast.
Asunto(s)
Hipersensibilidad a los Alimentos/prevención & control , Inflamación/prevención & control , Probióticos/administración & dosificación , Saccharomyces cerevisiae/fisiología , Administración Oral , Animales , Modelos Animales de Enfermedad , Femenino , Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/inmunología , Interleucina-17/sangre , Interleucina-17/inmunología , Ratones , Ratones Endogámicos BALB C , Viabilidad Microbiana , Peroxidasa/metabolismoRESUMEN
Inflammatory bowel diseases (IBD) are chronic processes involving a deregulated immune response against intestinal microbiota in genetically susceptible individuals. Ulcerative colitis (UC) is an IBD restricted to colonic mucosa and its chronicity is a predisposing factor for colorectal cancer (CRC). Probiotics have been investigated as an adjuvant treatment for UC, and Escherichia coli Nissle 1917 (EcN) was the focus of our investigation. The aim of this study was to investigate the preventive effect of the EcN probiotic in an experimental model of chronic colitis in germ-free (GF) and conventional (CV) mice. CV female mice were used for clinical, immunological and permeability experiments. GF mice were used for a faecal microbiota transplantation assay. To induce colitis, three cycles of 3.0% dextran sulphate sodium (DSS) were administered to the animals. For probiotic treatment, the mice received a daily intragastric gavage of 9.0 log10 cfu of EcN, beginning 10 days before colitis induction and continuing until the end of the experiment. EcN presented beneficial effects when administered preventively. Daily Disease Activity Index (DAI) evolution demonstrated significant difference in remission periods after the first two DSS cycles and during the third one. Reduction in bacterial translocation after probiotic treatment indicated protection of the intestinal barrier. Associated with mucosal preservation, restoration of secretory immunoglobulin A levels and reduction of interleukin (IL)-5, IL-13, tumour necrosis factor and interferon-γ levels were observed in EcN treatment. Finally, when microbiota modification was verified, 16S rRNA-based compositional analysis showed variation of intestinal microbiota between the control and colitis groups. After faecal transplantation using GF mice, it was observed that EcN treatment in CV mice might result in modulated intestinal microbiota. This was observed indirectly in the reduced daily DAI, when colitis was compared with treated group. In conclusion, EcN presented beneficial effects in this model, suggesting its usefulness for treating UC.
Asunto(s)
Colitis Ulcerosa/prevención & control , Escherichia coli/fisiología , Trasplante de Microbiota Fecal , Mucosa Intestinal/microbiología , Probióticos/farmacología , Animales , Colon/patología , Sulfato de Dextran/toxicidad , Modelos Animales de Enfermedad , Escherichia coli/clasificación , Femenino , Microbioma Gastrointestinal/fisiología , Vida Libre de Gérmenes , Inmunoglobulina A/análisis , Interferón gamma/sangre , Interleucina-13/sangre , Interleucina-5/sangre , Mucosa Intestinal/patología , Ratones , ARN Ribosómico 16S/genética , Factor de Necrosis Tumoral alfa/sangreRESUMEN
This study evaluated the effects of Bifidobacterium longum 51A on the intestinal mucosa and inflammatory response in experimental colitis. Colitis was induced by administration of 3.5% dextran sodium sulphate (DSS) solution for 7 days. Two periods of administration were performed: treatment (T) group, mice received Bifidobacterium only during disease induction (7 days); total treatment (TT) group, mice received Bifidobacterium for 10 days before and during disease induction. The probiotic effects on intestinal permeability, inflammatory infiltrate, histological analysis, cytokines, chemokines and sIgA were evaluated. Bifidobacterium administration in the T group showed reduction in intestinal permeability and lower IL-1ß, myeloperoxidase, and eosinophil peroxidase levels compared to those in the colitis group (P<0.05). Bifidobacterium administration in the TT group attenuated severe lesions in the colon and reduced eosinophil peroxidase level (P<0.05). B. longum 51A treatment modality was more effective than total treatment and reduced the inflammatory response and its consequences on intestinal epithelium.
Asunto(s)
Bifidobacterium longum , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Probióticos/uso terapéutico , Animales , Colitis/inducido químicamente , Colon/efectos de los fármacos , Colon/microbiología , Colon/patología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Peroxidasa del Eosinófilo/metabolismo , Femenino , Inmunoglobulina A Secretora/metabolismo , Inflamación/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/inmunología , Enfermedades Inflamatorias del Intestino/microbiología , Interleucina-1beta/metabolismo , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/patología , Intestinos/efectos de los fármacos , Intestinos/patología , Ratones , Ratones Endogámicos BALB C , Peroxidasa/metabolismoRESUMEN
The aim of the present study was to evaluate the effect of vascular endothelial growth factor (VEGF) on the survival and growth of goat preantral follicles after in vitro culture and to verify the expression of VEGF receptor (VEGFR)-2 in goat ovaries. Ovarian fragments were cultured for 1 or 7 days in minimal essential medium (MEM) with different concentrations of VEGF (1, 10, 50, 100 or 200 ng mL(-1)). Non-cultured (fresh control) and cultured tissues were processed for histological and ultrastructural studies. The results showed that 200 ng mL(-1) VEGF resulted in a similar percentage of normal preantral follicles after 1 and 7 days of culture compared with control. Compared with basic culture medium alone, an increase in follicular and oocyte diameters was observed in the presence of 10 ng mL(-1) VEGF after 7 days culture. Ultrastructural analysis confirmed follicular integrity after 7 days culture in the presence of 200 ng mL(-1) VEGF. Immunohistochemical studies demonstrated the expression of VEGFR-2 in oocytes and granulosa cells of all follicular stages, except in granulosa cells of primordial follicles. In conclusion, the present study has shown that VEGF maintains follicular ultrastructural integrity and promotes follicular growth. In addition, VEGFR-2 is expressed in oocytes of caprine ovarian follicles at all developmental stages and in granulosa cells of developing follicles.
Asunto(s)
Cabras/metabolismo , Folículo Ovárico/efectos de los fármacos , Ovario/metabolismo , Receptores de Factores de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Fase Folicular/fisiología , Células de la Granulosa/metabolismo , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/ultraestructura , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
AIMS: To compare the effects of five yeast probiotic formulations on viability, revival and washout kinetic in the digestive tract of mice, and the protection against an experimental infection with Salmonella enterica serovar Typhimurium. METHODS AND RESULTS: The number of viable cells in five commercial probiotic products codified as A, B, C and D (Saccharomyces boulardii- lyophilized) and E (Saccharomyces cerevisiae- aqueous suspension) was determined, as well as revival and washout kinetic in mouse intestine. Protective capacity was evaluated by survival rate and histopathology of liver and intestine of mice treated with each product and then challenged with Salm. Typhimurium. CONCLUSIONS: Product A contained the highest number of viable cells and, fed to mice, gave the highest counts of viable yeasts and the longest persistence in faeces. Probably as a consequence, the highest survival and protection of intestinal and hepatic tissues were observed when product A was used for mouse treatment. Product E showed low counts in the formulation and was not recovered from mouse intestine. SIGNIFICANCE AND IMPACT OF THE STUDY: Formulation (lyophilization or aqueous suspension) is an important factor for revival and survival of a probiotic product in vivo and consequently for its protective properties.
Asunto(s)
Probióticos/administración & dosificación , Probióticos/farmacología , Saccharomyces , Salmonelosis Animal/prevención & control , Animales , Recuento de Colonia Microbiana , Heces/microbiología , Intestinos/microbiología , Intestinos/patología , Hígado/patología , Ratones , Salmonelosis Animal/patología , Salmonella typhimuriumRESUMEN
The aim of the present study was to investigate the effects of growth and differentiation factor-9 (GDF-9) on the survival and activation of preantral follicles, as well as their subsequent progression to secondary follicles, using goat ovarian cortical culture in vitro. Pieces of ovarian cortex were cultured for 1 and 7 days in minimum essential medium (MEM) with or without different concentrations of GDF-9 (1-200 ng mL(-1)). On Day 0 and after 1 and 7 days of culture, cortical pieces were fixed for histological and transmission electron microscopy evaluation. Preantral follicles were classified according to their development stage (primordial, intermediate, primary and secondary) and on the basis of morphological features (normal or degenerated). In addition, follicular and oocyte diameters were determined before and after culture. The results showed that, compared with non-cultured cortical tissue (Day 0), the culture of ovarian tissue significantly reduced (P < 0.05) the percentage of normal follicles in all media tested, except for tissue cultured in the presence of 200 ng mL(-1) GDF-9. Furthermore, in all media tested, the percentage of primordial follicles was significantly reduced (P < 0.05), with a concomitant increase in the percentage of developing follicles. The highest percentage of secondary follicles was observed after 7 days of culture in MEM plus 200 ng mL(-1) GDF-9. At all concentrations of GDF-9 tested, follicular diameter increased significantly after 7 days of culture compared with non-cultured cortical tissue. In conclusion, the results of the present study indicate that 200 ng mL(-1) GDF-9 maintains the survival of preantral follicles and promotes activation of primordial follicles. Furthermore, GDF-9 stimulates the transition from primary to secondary follicles, maintaining ultrastructural integrity of the follicles.
Asunto(s)
Factor 9 de Diferenciación de Crecimiento/farmacología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/crecimiento & desarrollo , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Cabras , Folículo Ovárico/citologíaRESUMEN
The aim of the present study was to evaluate the effects of storage of goat ovarian fragments at different temperatures and for different incubation times on the viability and growth of cultured preantral follicles in vitro. Caprine ovaries were collected and divided into 19 fragments, with one fragment being fixed immediately (fresh control). The remaining fragments were placed in minimal essential medium (MEM) and maintained at 4, 20 or 35 degrees C for 2 or 4 h. After each incubation period, some of the fragments were fixed (non-cultured), whereas others were cultured in vitro for 1 or 7 days. Fragments were processed to enable routine histological and transmission electron microscopic examination. After 7 days of culture, only ovarian fragments stored at 4 degrees C for 4 h maintained a percentage of morphologically normal follicles similar to that in the fresh control. For all other treatments groups, there was a significant increase in follicular activation observed. In addition, there was an increase in oocyte and follicular diameter after culture of ovarian cortex that had been chilled previously at 4 degrees C for 2 or 4 h. In conclusion, the present study demonstrated that chilling ovarian fragments at 4 degrees C during transportation is best for maintaining follicle viability and to increase follicular growth during in vitro culture.
Asunto(s)
Frío , Cabras/fisiología , Recuperación del Oocito/métodos , Oocitos/crecimiento & desarrollo , Folículo Ovárico/fisiología , Transportes , Animales , Tamaño de la Célula , Supervivencia Celular , Células Cultivadas , Femenino , Oocitos/citología , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/ultraestructura , Ovario/fisiología , TemperaturaRESUMEN
Allergic asthma is a chronic disease mainly characterised by eosinophil inflammation and airway remodelling. Many studies have shown that the gut microbiota of allergic individuals differs from that of non-allergic individuals. Although high levels of bifidobacteria have been associated with healthy persons, Bifidobacterium adolescentis ATCC 15703, a gut bacteria, has been associated with allergic individuals in some clinical studies. The relationship between B. adolescentis ATCC 15703 and asthma or allergies has not been well elucidated, and its effect may be dependent on the host's genetic profile or disease state. To elucidate this question, we evaluated the role of preventive B. adolescentis ATCC 15703 treatment on experimental allergic airway inflammation in two genetically different mouse strains, Balb/c and C57BL/6 (B6). Balb/c mice display a greater predisposition to develop allergic responses than B6 mice. Oral preventive treatment with B. adolescentis ATCC 15703 modulated experimental allergic airway inflammation, specifically in Balb/c mice, which showed decreased levels of eosinophils in the airway. B6 mice did not exhibit any significant alterations in eosinophils but showed an increased influx of total leukocytes and neutrophils into the airway. The mechanism underlying the beneficial effects of these bacteria in experimental allergic mice may involve products of bacteria metabolism, as dead bacteria did not mimic the ability of live B. adolescentis ATCC 15703 to attenuate the influx of eosinophils into the airway. To conclude, preventive oral B. adolescentis ATCC 15703 treatment can attenuate the major characteristic of allergic asthma, eosinophil airway influx, in Balb/c but not B6 mice. These results suggest that oral treatment with this specific live bacterial strain may have therapeutic potential for the treatment of allergic airway disease, although its effect is mouse-strain-dependent.
Asunto(s)
Asma/prevención & control , Bifidobacterium adolescentis/crecimiento & desarrollo , Probióticos/administración & dosificación , Sistema Respiratorio/patología , Administración Oral , Animales , Modelos Animales de Enfermedad , Eosinófilos/inmunología , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Resultado del TratamientoRESUMEN
The use of probiotics to prevent or treat mucosal inflammation has been studied; however, the combined effect of probiotics and prebiotics is unclear. The aim of this study was to test whether oral administration of a synbiotic (Simbioflora®) preparation containing Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium lactis plus fructooligosaccharide could help control mucosal inflammation in experimental mucositis induced by 5-fluorouracil (5-FU). Male BALB/c mice were randomly divided into six groups: control (CTL), control + prebiotic (CTL+P), control + synbiotic (CTL+S), mucositis (MUC), mucositis + prebiotic (MUC+P), and mucositis + synbiotic (MUC+S). Mice from the CTL+S, MUC+S, CTL+P, and MUC+P groups received synbiotic or prebiotic daily by oral gavage for 13 days. Mice in the CTL and MUC groups received the same volume of saline. On day 11, mice in the MUC, MUC+P, and MUC+S groups received an intraperitoneal injection of 300 mg/kg 5-FU to induce mucositis. After 72 h, all mice were euthanised. Intestinal permeability, intestinal histology, and biochemical parameters were analysed. Group MUC showed a greater weight loss and increased intestinal permeability (0.020 counts per min [cpm]/g) compared to the CTL group (0.01 cpm/g) P<0.05. Both treatments attenuated weight loss compared to the MUC group. Nonetheless, the synbiotic caused a greater reduction in intestinal permeability (0.012 cpm/g) compared to the MUC (0.020 cpm/g) and MUC+P (0.016 cpm/g) groups P<0.05. Mice in groups MUC+P and MUC+S displayed significant recovery of lesions and maintenance of the mucus layer. There were no differences in the short-chain fatty acid concentrations in the faeces between the MUC and CTL groups (P>0.05). Increased acetate and propionate concentrations were evidenced in the faeces of the MUC+P and MUC+S groups. Only the synbiotic treatment increased the butyrate concentration (P<0.05). The results indicate that administration of synbiotic can decrease mucosal damage caused by mucositis.
Asunto(s)
Mucositis/prevención & control , Simbióticos/administración & dosificación , Administración Oral , Animales , Bifidobacterium animalis/crecimiento & desarrollo , Bifidobacterium animalis/metabolismo , Peso Corporal , Ácidos Grasos Volátiles/análisis , Heces/química , Fluorouracilo/administración & dosificación , Fluorouracilo/toxicidad , Tracto Gastrointestinal/patología , Lactobacillus/crecimiento & desarrollo , Lactobacillus/metabolismo , Ratones Endogámicos BALB C , Mucositis/inducido químicamente , Oligosacáridos/administración & dosificación , Oligosacáridos/metabolismo , Resultado del TratamientoRESUMEN
The aim of the study was to assess the efficacy of Saccharomyces boulardii in experimental treatment of giardiasis and its impact on intestinal integrity and some functions of gerbils infected with Giardia lamblia. 28 gerbils (Meriones unguiculatus), aged 4-6 weeks, were divided into four groups: untreated and uninfected control (CT); infected with G. lamblia (IGL); treated with S. boulardii (SB); and infected with G. lamblia and treated with S. boulardii (ITSB). The SB and ITSB groups received S. boulardii 15 days prior to being infected with G. lamblia. The treatment continued until completion of the experiment (22nd day). The IGL and ITSB groups were gavage-inoculated with G. lamblia ensuring one-week infection. 4 h before euthanasia, all animals were gavaged with a solution containing diethylenetriamine-pentaacetic acid (DTPA) marked with technetium-99mTc DTPA to determine intestinal permeability. The small intestine was removed for histopathological, morphometric analysis and count of trophozoites adhered to the mucosa. The selected probiotic caused an approximate reduction of 70% of parasite load, which was determined by attached trophozoites (P<0.01) and immune-marked trophozoites (P<0.05). Treatment with S. boulardii (SB and ITSB groups) also increased the height of the intestinal villi and crypt depth compared to the CT and IGL groups (P<0.05). The area of mucus production and the number of goblet cells of the SB and ITSB groups were higher compared to the CT and IGL groups (P<0.01). The animals treated with S. boulardii also exhibited a significant increase of intraepithelial lymphocytes counts (P<0.01). There was no difference in the intestinal permeability between the groups studied. The efficacy of S. boulardii in reducing damages caused by Giardia was demonstrated, with an approximate reduction of 70% of the parasite load, suggesting its use as a coadjuvant in giardiasis treatment.
Asunto(s)
Giardia lamblia/fisiología , Giardiasis/tratamiento farmacológico , Probióticos/administración & dosificación , Saccharomyces boulardii/fisiología , Animales , Modelos Animales de Enfermedad , Gerbillinae , Giardia lamblia/efectos de los fármacos , Giardia lamblia/crecimiento & desarrollo , Giardiasis/parasitología , Humanos , Intestino Delgado/parasitología , Intestino Delgado/patología , MasculinoRESUMEN
STING (stimulator of interferon genes) is a cytosolic sensor for cyclic dinucleotides and also an adaptor molecule for intracellular DNA receptors. Although STING has important functions in the host defense against pathogens and in autoimmune diseases, its physiological relevance in intestinal homeostasis is largely unknown. In this study, we show that STING-/- mice presented defective protective mechanisms of intestinal mucosa, including decreased number of goblet cells, diminished mucus production, and lower levels of secretory IgA, when compared with wild-type (WT) mice. Fecal content and microbiota DNA could activate STING, indicating a role of this molecule in gut. Microbiota composition was altered in STING-/- mice toward a more inflammatory profile, evidencing a reduction in the Allobacolum and Bifidobacterium groups along with increase in Disulfovibrio bacteria. Absence of STING lead to decrease in induced intraepithelial lymphocytes (IEL) and to increase in group 1 innate lymphoid cell (ILC1) as well as ILC3 frequencies and decrease in ILC2 in the colon. Development and function of Foxp3+ and LAP+ regulatory T cells were also compromised in STING-/- mice. Moreover, these mice were highly susceptible to dextran sodium sulfate-induced colitis, T-cell-induced colitis, and enteric Salmonella typhimurium infection when compared with WT animals. Therefore, our results identify an important role of STING in maintaining gut homeostasis and also a protective effect in controlling gut inflammation.
Asunto(s)
Colitis/inmunología , Microbioma Gastrointestinal/fisiología , Mucosa Intestinal/inmunología , Intestinos/fisiología , Linfocitos/inmunología , Proteínas de la Membrana/metabolismo , Infecciones por Salmonella/inmunología , Salmonella typhimurium/inmunología , Linfocitos T Reguladores/inmunología , Animales , Colitis/inducido químicamente , Colitis/genética , Sulfato de Dextran , Femenino , Factores de Transcripción Forkhead/metabolismo , Homeostasis , Inmunidad Innata , Inmunoglobulina A Secretora/sangre , Masculino , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infecciones por Salmonella/genética , Células TH1/inmunologíaRESUMEN
The aims of the present study were to evaluate the effects of fibroblast growth factor-2 (FGF-2) on survival, activation and growth of caprine early-staged (preantral) follicles using histological and ultrastructural studies. Fragments of caprine ovarian cortex were cultured for 1 or 5 days in an enriched minimum essential medium, supplemented or not with different concentrations of FGF-2 (10, 50 or 100 ng/ml). Fragments from non-cultured ovarian tissue (control) and from tissues cultured for 1 or 5 days in a specific medium were processed for transmission electron microscopy (TEM) or classical histology to evaluate the morphological quality of caprine preantral follicles and to calculate the percentages of normal follicles. Additionally, effects of FGF-2 on oocyte and follicle diameter of cultured preantral follicles were investigated. Our results showed that, although the percentages of histologically normal follicles were lower in cultured than in non-cultured ovarian tissue fragments, there were no differences in this regard among treatments, neither on day 1 nor on day 5 of culture. After 1 and 5 days of culture, a significantly higher percentage of growing follicles was observed in the medium supplemented with 50 ng/ml of FGF-2. This FGF-2 treatment furthermore resulted in an increase in diameter of both oocytes and follicles that were cultured for 5 days. TEM showed that the ultrastructural integrity of caprine preantral follicles was maintained during their 5-day culture in the presence of 50 ng/ml FGF-2. In conclusion, this study demonstrated that at a concentration of 50 ng/ml FGF-2 not only maintains the morphological integrity of caprine preantral follicles cultured for 5 days, but also stimulates the activation of primordial follicles and the growth of activated follicles.
Asunto(s)
Factor 2 de Crecimiento de Fibroblastos/farmacología , Cabras/fisiología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/crecimiento & desarrollo , Animales , Recuento de Células , Técnicas de Cultivo de Célula , Tamaño de la Célula , Medios de Cultivo/química , Relación Dosis-Respuesta a Droga , Femenino , Oocitos/fisiología , Folículo Ovárico/ultraestructura , Factores de TiempoRESUMEN
The aims of the present study were to investigate the effects of the interaction between follicle stimulating hormone (FSH) and fibroblast growth factor-2 (FGF-2) on survival, follicular growth initiation and further growth of caprine preantral follicles. Pieces of caprine ovarian cortex were cultured for 1 or 7 days in minimum essential medium (MEM) supplemented with FSH, FGF-2 or FSH + FGF-2. Small fragments from non-cultured ovarian tissue and from those cultured for 1 or 7 days were processed for classical histology and transmission electron microscopy (TEM) to verify follicular morphology and growth. The results showed that, after 7 days culture, the highest percentages of normal follicles were observed in medium supplemented with FSH. After 7 days culture, the interaction between FSH and FGF-2 was most effective to promote the initiation of primordial follicles growth and oocyte growth. TEM showed ultrastructural integrity of follicles after 1 day of culture in MEM and after 7 days in all treatments, except in those follicles cultured for 7 days in MEM. In conclusion, this study demonstrated that the interaction between FSH and FGF-2 stimulates the initiation of primordial follicles growth and the subsequent growth of developing follicles. Furthermore, these data showed that FSH is important to maintain follicular integrity after 7 days culture.