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1.
Int Immunopharmacol ; 130: 111638, 2024 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-38373387

RESUMEN

L-arginine, as an essential substance of the immune system, plays a vital role in innate immunity. MiR155, a multi-functional microRNA, has gained importance as a regulator of homeostasis in immune cells. However, the immunoregulatory mechanism between L-arginine and miR155 in bacterial infections is unknown. Here, we investigated the potential role of miR155 in inflammation and the molecular regulatory mechanisms of L-arginine in Streptococcus uberis (S. uberis) infections. And we observed that miR155 was up-regulated after infection, accompanying the depletion of L-arginine, leading to metabolic disorders of amino acids and severe tissue damage. Mechanically, the upregulated miR155 mediated by the p65 protein played a pro-inflammatory role by suppressing the suppressor of cytokine signaling 6 (SOCS6)-mediated p65 ubiquitination and degradation. This culminated in a violently inflammatory response and tissue damage. Interestingly, a significant anti-inflammatory effect was revealed in L-arginine supplementation by reducing miR155 production via inhibiting p65. This work firstly uncovers the pro-inflammatory role of miR155 and an anti-inflammatory mechanism of L-arginine in S.uberis infection with a mouse mastitis model. Collectively, we provide new insights and strategies for the prevention and control of this important pathogen, which is of great significance for ensuring human food health and safety.


Asunto(s)
Arginina , Mastitis , MicroARNs , Infecciones Estreptocócicas , Animales , Femenino , Humanos , Ratones , Arginina/metabolismo , Inflamación/metabolismo , MicroARNs/genética , Infecciones Estreptocócicas/metabolismo , Streptococcus/fisiología , Proteínas Supresoras de la Señalización de Citocinas/metabolismo , Mastitis/inmunología , Mastitis/metabolismo
2.
Oxid Med Cell Longev ; 2021: 5048375, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34938382

RESUMEN

Mastitis is mainly induced by gram-negative bacterial infections, causing devastating economic losses to the global cattle industry. Both selenium (Se) and taurine (Tau) exhibit multiple biological effects, including reducing inflammation. However, no studies have reported the protective effect of the combined use of Se and Tau against mastitis, and the underlying mechanisms remain unclear. In this study, lipopolysaccharide (LPS), the vital virulence factor of gram-negative bacteria, was used to construct the in vivo and vitro mastitis models. The results of in vivo model showed that Se and Tau combination was more effective than either substance alone in reducing tissue hyperemia, edema, and neutrophil infiltration in the mammary acinar cavity, improving the blood-milk barrier in LPS-induced mice mastitis, and decreasing the expression of proinflammatory factors and the activity of MPO. Moreover, Se and Tau combination significantly increased the levels of LPS-induced reduction in PI3K/Akt/mTOR, but the expressions of TLRs and NLRP3 were not significantly changed in the mammary tissue. In the in vitro experiments, the effects of Se and Tau combination or alone on inflammatory factors, inflammatory mediators, MPO activity, and blood-milk barrier were consistent with those in vivo. The Se and Tau combination has also been found to increase the survival rate of BMECs compared with each substance alone via promoting cellular proliferation and inhibiting apoptosis. Also, it has been confirmed that this combination could restore the LPS-induced inhibition in the PI3K/Akt/mTOR signaling pathway. Inhibition of mTOR by Rapamycin counteracted the combined protection of SeMet and Tau against LPS-induced inflammatory damage, the inhibition of PI3K by LY294002 blocked the activation of mTOR, and the accumulation of ROS by the ROS agonist blocked the activation of PI3K. In conclusion, these findings suggested that Se and Tau combination was better than either substance alone in protecting LPS-induced mammary inflammatory lesions by upregulating the PI3K/Akt/mTOR signaling pathway.


Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Inflamación/prevención & control , Glándulas Mamarias Animales/efectos de los fármacos , Mastitis/prevención & control , Especies Reactivas de Oxígeno/metabolismo , Selenio/farmacología , Taurina/farmacología , Animales , Antiinflamatorios/farmacología , Bovinos , Quimioterapia Combinada , Femenino , Depuradores de Radicales Libres , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/metabolismo , Lipopolisacáridos/toxicidad , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/metabolismo , Glándulas Mamarias Animales/patología , Mastitis/inducido químicamente , Mastitis/inmunología , Mastitis/metabolismo , Ratones , Ratones Endogámicos ICR , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo
3.
Food Funct ; 10(10): 6543-6555, 2019 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-31545328

RESUMEN

Mastitis, a major disease affecting dairy cows, is most commonly caused by Staphylococcus aureus (S. aureus). Selenium (Se) can activate pivotal proteins in immune responses and regulate the immune system, and microRNA-155 (miR-155) is a key transcriptional regulator for inflammation-related diseases. We constructed the model of mouse mastitis in vivo and primary mouse mammary epithelial cells (MMECs) in vitro, which were induced by S. aureus. Se content of the mammary was estimated using an atomic fluorescence spectrophotometer. Histopathological analysis was performed via hematoxylin and eosin (H&E) staining. The mmu-miR-155-5p mimic was transfected in MMECs, and viability was determined through the MTT assay. Transfected efficiency was evaluated by qPCR and fluorescence staining. Cytokines including TNF-α, IL-1ß, IL-10 and TLRs were detected with qPCR. In addition, western blotting was used to evaluate the expression of the NF-κB and MAPKs signaling pathways. The results demonstrated that a Se-supplemented diet improved the content of Se in mammary tissues. Histopathological studies indicated that the mammary glands were protected in the Se-supplemented group after S. aureus infection. Se-supplementation suppressed the production of MPO, mmu-miR-155, TNF-α, IL-1ß, and TLR2 and significantly inhibited the phosphorylation of NF-κB and MAPKs in vivo and in vitro. All the data indicated that mmu-miR-155 played a pro-inflammatory role in our study, and Se-supplementation could suppress the expression of mmu-miR-155 to inhibit inflammation in S. aureus-induced mastitis in mice.


Asunto(s)
Enfermedades de los Bovinos/tratamiento farmacológico , Mastitis/tratamiento farmacológico , MicroARNs/genética , Selenio/administración & dosificación , Infecciones Estafilocócicas/veterinaria , Animales , Bovinos , Enfermedades de los Bovinos/genética , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/microbiología , Citocinas/genética , Citocinas/inmunología , Femenino , Regulación de la Expresión Génica , Mastitis/genética , Mastitis/inmunología , Mastitis/microbiología , Ratones , MicroARNs/inmunología , FN-kappa B/genética , FN-kappa B/inmunología , Infecciones Estafilocócicas/genética , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/fisiología , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/inmunología
4.
Biol Trace Elem Res ; 191(1): 159-166, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-30523572

RESUMEN

Mastitis is one of the most important diseases affecting the dairy industry in the world, and it also poses a great threat to human food safety. In this study, we explored whether selenium can inhibit the activation of the NALP3 inflammasome and NF-κB/MAPK pathway to achieve anti-inflammatory effects. Sixty BALB/c female mice were randomly divided into three groups according to diets of different selenium concentrations (high, normal, and low). After 90 days, mice fed the same selenium concentration were randomly divided into two smaller groups, one of which was inoculated with Staphylococcus aureus and the other injected with saline as a control. Through histopathologic examination staining, western blot, qPCR, and ELISA, the results showed that with increasing selenium concentrations, the expression levels of IL-1ß, TNF-α, NALP3, caspase-1, and ASC were decreased in mouse mammary tissue. Therefore, this study revealed that selenium can attenuate S. aureus mastitis by inhibiting the activation of the NALP3 inflammasome and NF-κB/MAPK pathway.


Asunto(s)
Inflamasomas/inmunología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Mastitis/inmunología , FN-kappa B/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Selenio/farmacología , Infecciones Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Animales , Femenino , Sistema de Señalización de MAP Quinasas/inmunología , Mastitis/microbiología , Mastitis/patología , Ratones , Ratones Endogámicos BALB C , Infecciones Estafilocócicas/patología
5.
Front Immunol ; 9: 2884, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30574152

RESUMEN

A novel vaccine against bovine viral diarrhea (BVD) induced pathogenic antibody production in 5-10% of BVD-vaccinated cows. Transfer of these antibodies via colostrum caused Bovine neonatal pancytopenia (BNP) in calves, with a lethality rate of 90%. The exact immunological mechanisms behind the onset of BNP are not fully understood to date. To gain further insight into these mechanisms, we analyzed the immune proteome from alloreactive antibody producers (BNP cows) and non-responders. After in vitro stimulation of peripheral blood derived lymphocytes (PBL), we detected distinctly deviant expression levels of several master regulators of immune responses in BNP cells, pointing to a changed immune phenotype with severe dysregulation of immune response in BNP cows. Interestingly, we also found this response pattern in 22% of non-BVD-vaccinated cows, indicating a genetic predisposition of this immune deviant (ID) phenotype in cattle. We additionally analyzed the functional correlation of the ID phenotype with 10 health parameters and 6 diseases in a retrospective study over 38 months. The significantly increased prevalence of mastitis among ID cows emphasizes the clinical relevance of this deviant immune response and its potential impact on the ability to fight infections.


Asunto(s)
Animales Recién Nacidos/inmunología , Diarrea Mucosa Bovina Viral/prevención & control , Mastitis/inmunología , Pancitopenia/inmunología , Vacunas Virales/efectos adversos , Crianza de Animales Domésticos , Animales , Animales Recién Nacidos/sangre , Antígenos Virales/inmunología , Diarrea Mucosa Bovina Viral/virología , Bovinos , Calostro/inmunología , Calostro/metabolismo , Virus de la Diarrea Viral Bovina/inmunología , Femenino , Incidencia , Isoanticuerpos/inmunología , Isoanticuerpos/metabolismo , Isoantígenos/inmunología , Linfocitos , Mastitis/epidemiología , Pancitopenia/mortalidad , Pancitopenia/veterinaria , Fenotipo , Embarazo , Estudios Retrospectivos , Vacunación/efectos adversos , Vacunas Virales/administración & dosificación
6.
Microb Pathog ; 117: 341-347, 2018 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-29510207

RESUMEN

The predominant Staphylococcus aureus (S. aureus), an etiological agent of camel mastitis is becoming drug resistant that invites prevention and control strategies. Vaccine production would have a valuable impact on public health. Therefore, in present study, inactivated vaccine with different adjuvants was prepared and evaluated against S. aureus. The vaccinal isolate recovered from camel subclinical mastitis was coagulase positive (PCR based), having expressed pseudocapsule, holding alpha-beta hemolysin characteristics, and multiple drug resistant. Inactivated alum precipitated S. aureus vaccine (APSV) and oil adjuvant S. aureus vaccine (OASV) were prepared after confirming its antigenicity in rabbits. Three groups of rabbits were randomly inoculated with APSV, OASV, and placebo (Unvaccinated, UV). Each group was further divided into two groups based on single and booster dose inoculation. Booster dose of vaccines in rabbits at day 15th of primary inoculation was given. Serum samples were taken on 15, 30, 45 and 60 days of primary inoculation from all rabbits. Analysis of variance was applied to compare geometric mean titer (GMT) of three groups, while t-test was applied to estimate the difference between single and booster dose response. The study found 1010 CFU/mL S. aureus as standard bacterial load for vaccines with higher and sustained antigenicity. The vaccines were safe from morbidity and mortality, and proved effective and stable for 7 and 4 months at 25 °C and 37 °C, respectively. The OASV produced significantly (p < 0.05) higher immune response followed by APSV throughout trial. The highest GMT by APSV and OASV vaccines with single dose inoculation was 37.92 and 69.92 at day 45th post primary inoculation, respectively. Similarly, 59.20 and 142.40 GMTs were noted with booster dose in case of APSV and OASV, respectively. The booster dose presented significantly (p < 0.05) higher GMT than that of single dose inoculation of vaccines. The study concluded APSV and OASV safe, effective, and stable with significant immunogenic results in experimental rabbits.


Asunto(s)
Inmunogenicidad Vacunal/inmunología , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/prevención & control , Vacunas Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Vacunación , Vacunas de Productos Inactivados/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Compuestos de Alumbre , Animales , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/inmunología , Carga Bacteriana , Proteínas Bacterianas/inmunología , Camelus , Coagulasa , Modelos Animales de Enfermedad , Farmacorresistencia Bacteriana Múltiple , Femenino , Proteínas Hemolisinas , Inmunización Secundaria , Mastitis/inmunología , Mastitis/microbiología , Mastitis/prevención & control , Aceite Mineral/administración & dosificación , Conejos , Infecciones Estafilocócicas/microbiología , Vacunas Estafilocócicas/administración & dosificación , Staphylococcus aureus/patogenicidad , Factores de Tiempo , Vacunas de Productos Inactivados/administración & dosificación
7.
J Microbiol Biotechnol ; 26(3): 579-87, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26608166

RESUMEN

Mastitis is a prevalent inflammatory disease that remains one of the main causes of poor quality of milk. Phytoncides are naturally occurring anti-inflammatory compounds derived from plants and trees. To determine if treatment with phytoncide could decrease the severity of lipopolysaccharide (LPS)-induced inflammatory responses, mammary alveolar epithelial cells (MAC-T) were pretreated with phytoncide (0.02% and 0.04% (v/v)) followed by LPS treatment (1 and 25 µg/ml). The results demonstrated that phytoncide downregulated LPS-induced pro-inflammatory cyclooxygenase-2 (COX-2) expression. Additionally, LPS-induced activation of ERK1/2, p38, and Akt was attenuated by phytoncide. Treatment of cells with known pharmacological inhibitors of ERK1/2 (PD98059), p38 (SB203580), and Akt (LY294002) confirmed the association of these signaling pathways with the observed alterations in COX-2 expression. Moreover, phytoncide attenuated LPS-induced NF-κB activation and superoxide production, and, finally, treatment with phytoncide increased Nrf2 activation. Results suggest that phytoncide can decrease LPS-induced inflammation in MAC-T cells.


Asunto(s)
Antiinflamatorios/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/inmunología , Lipopolisacáridos/inmunología , Mastitis/inmunología , Pinus/química , Extractos Vegetales/farmacología , Animales , Bovinos , Ciclooxigenasa 2 , Femenino , Frutas/química , Mastitis/tratamiento farmacológico , Mastitis/genética , FN-kappa B/genética , FN-kappa B/inmunología , Extractos Vegetales/aislamiento & purificación , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología , Compuestos Orgánicos Volátiles/aislamiento & purificación , Compuestos Orgánicos Volátiles/farmacología
8.
Int Immunopharmacol ; 27(1): 130-7, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25939535

RESUMEN

Mastitis, which commonly occurs during the postpartum period, is caused by the infection of the mammary glands. The most common infectious bacterial pathogen of mastitis is Staphylococcus aureus (S. aureus) in both human and animals. Brazilin, a compound isolated from the traditional herbal medicine Caesalpinia sappan L., has been shown to exhibit multiple biological properties. The present study was performed to determine the effect of brazilin on the inflammatory response in the mouse model of S. aureus mastitis and to confirm the mechanism of action involved. Brazilin treatment was applied in both a mouse model and cells. After brazilin treatment of cells, Western blotting and qPCR were performed to detect the protein levels and mRNA levels, respectively. Brazilin treatment significantly attenuated inflammatory cell infiltration and inhibited the expressions of TNF-α, IL-1ß and IL-6 in a dose-dependent manner. Administration of brazilin in mice suppressed S. aureus-induced inflammatory injury and the production of proinflammatory mediators. This suppression was achieved by reducing the increased expression of TLR2 and regulating the NF-κB and MAPK signaling pathways in the mammary gland tissues and cells with S. aureus-induced mastitis. These results suggest that brazilin appears to be an effective drug for the treatment of mastitis and may be applied as a clinical therapy.


Asunto(s)
Benzopiranos/administración & dosificación , Mastitis/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/inmunología , Receptor Toll-Like 2/metabolismo , Animales , Caesalpinia/inmunología , Células Cultivadas , Citocinas/metabolismo , Quinasas MAP Reguladas por Señal Extracelular , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Mediadores de Inflamación/metabolismo , Mastitis/inmunología , Ratones , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Infecciones Estafilocócicas/inmunología , Receptor Toll-Like 2/genética
9.
Inflammation ; 38(3): 1142-50, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25487780

RESUMEN

Mastitis is a major disease in humans and other animals and is characterized by mammary gland inflammation. It is a major disease of the dairy industry. Bergenin is an active constituent of the plants of genus Bergenia. Research indicates that bergenin has multiple biological activities, including anti-inflammatory and immunomodulatory properties. The objective of this study was to evaluate the protective effects and mechanism of bergenin on the mammary glands during lipopolysaccharide (LPS)-induced mastitis. In this study, mice were treated with LPS to induce mammary gland mastitis as a model for the disease. Bergenin treatment was initiated after LPS stimulation for 24 h. The results indicated that bergenin attenuated inflammatory cell infiltration and decreased the concentration of NO, TNF-α, IL-1ß, and IL-6, which were increased in LPS-induced mouse mastitis. Furthermore, bergenin downregulated the phosphorylation of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinases (MAPK) signaling pathway proteins in mammary glands with mastitis. In conclusion, bergenin reduced the expression of NO, TNF-α, IL-1ß, and IL-6 proinflammatory cytokines by inhibiting the activation of the NF-κB and MAPKs signaling pathways, and it may represent a novel treatment strategy for mastitis.


Asunto(s)
Antiinflamatorios/uso terapéutico , Benzopiranos/uso terapéutico , Mastitis/tratamiento farmacológico , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Animales , Modelos Animales de Enfermedad , Femenino , Inflamación/tratamiento farmacológico , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Lipopolisacáridos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/patología , Mastitis/inmunología , Mastitis/patología , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/metabolismo , Fosforilación/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Factor de Necrosis Tumoral alfa/metabolismo
10.
Life Sci ; 119(1-2): 9-17, 2014 Dec 05.
Artículo en Inglés | MEDLINE | ID: mdl-25445441

RESUMEN

AIMS: Geniposide, a major iridoid glycoside found in gardenia fruit, is widely used in Asian countries for its anti-inflammatory, anti-tumor and anti-apoptotic activities. Although the anti-inflammatory effect of geniposide has been widely reported, its anti-apoptotic role in mastitis remains unclear. In the present study, we investigated whether geniposide exerts anti-apoptotic activity in lipopolysaccharide (LPS)-induced mouse mammary glands. MAIN METHODS: We established a LPS-induced mouse mastitis model and LPS-stimulated primary mouse mammary epithelial cells (mMECs) model to investigate the anti-apoptotic effect of geniposide and the underlying mechanism of action. In the in vivo studies, apoptosis in mammary glands was detected by TUNEL. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the expression of Bax, Bcl-2, Caspase-3 and p53. In the in vitro study, the apoptosis in mammary epithelial cells was measured by Live-Dead staining. Western blot and qRT-PCR analysis were used to analyze the expression of Bax, Bcl-2, Caspase-3, p53 and TLR4. KEY FINDINGS: Geniposide alleviated mammary gland apoptosis, down-regulated Bax expression, inhibited Caspase-3 cleavage and p53 phosphorylation and up-regulated Bcl-2 expression in vivo. In vitro, geniposide decreased the ratio of dead cells in a dose-dependent manner. Geniposide inhibited Bax expression and Caspase-3 cleavage, and up-regulated the expression of Bcl-2. Moreover, geniposide down-regulated the expression of TLR4 and repressed the phosphorylation of p53. SIGNIFICANCE: These results demonstrate that the anti-apoptotic property of geniposide is due to its modulation of TLR4 and apoptosis-related factors (p53, Bax, Bcl-2 and Caspase-3) in LPS-induced mouse mastitis.


Asunto(s)
Antiinflamatorios/uso terapéutico , Apoptosis/efectos de los fármacos , Iridoides/uso terapéutico , Glándulas Mamarias Animales/efectos de los fármacos , Mastitis/tratamiento farmacológico , Receptor Toll-Like 4/inmunología , Animales , Antiinflamatorios/química , Células Cultivadas , Femenino , Gardenia/química , Iridoides/química , Lipopolisacáridos , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/patología , Mastitis/inducido químicamente , Mastitis/inmunología , Mastitis/patología , Ratones , Ratones Endogámicos BALB C
11.
J Surg Res ; 192(2): 573-81, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24972733

RESUMEN

BACKGROUND: Tea brewed from the leaves of persimmon or Rosa agrestis have several medical functions including treating allergy, antiatopic dermatitis, and anti-inflammatory effects. The objective of this study was to investigate the molecular mechanisms of astragalin, a main flavonoid component isolated from these herbs, in modifying lipopolysaccharide (LPS)-induced signaling pathways in primary cultured mouse mammary epithelial cells (mMECs). MATERIALS AND METHODS: The mMECs were treated with LPS in the absence or presence of different concentrations of astragalin. The expression of proinflammatory cytokines tumor necrosis factor α, and interleukin 6, as well as nitric oxide production were determined by enzyme-linked immunosorbent assay and Griess reaction, respectively. Cyclooxygenase-2, inducible nitric oxide synthase, toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), inhibitor protein of NF-κB (IκBα), P38, extracellular signal-regulated kinase, and c-Jun N-terminal kinase were measured by Western blot. RESULTS: The results showed that astragalin suppressed the expression of tumor necrosis factor α, interleukin 6, and nitric oxide in a dose-dependent manner in mMECs. Western blot results showed that the expression of inducible nitric oxide synthase and cyclooxygenase-2 was inhibited by astragalin. Besides, astragalin efficiently decreased LPS-induced TLR4 expression, NF-κB activation, IκBα degradation, and the phosphorylation of p38, extracellular signal-regulated kinase in BMECs. CONCLUSIONS: Our results indicated that astragalin exerts anti-inflammatory properties possibly via the inactivation of TLR4-mediated NF-κB and mitogen-activated protein kinases signaling pathways in LPS-stimulated mMECs. Thus, astragalin may be a potential therapeutic agent for bovine mastitis.


Asunto(s)
Antiinflamatorios/farmacología , Quempferoles/farmacología , Glándulas Mamarias Animales/efectos de los fármacos , Mastitis/tratamiento farmacológico , Preparaciones de Plantas/farmacología , Animales , Ciclooxigenasa 2/metabolismo , Diospyros/química , Interacciones Farmacológicas , Femenino , Interleucina-6/metabolismo , Quempferoles/química , Lipopolisacáridos/farmacología , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/inmunología , Mastitis/inducido químicamente , Mastitis/inmunología , Ratones Endogámicos BALB C , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Preparaciones de Plantas/química , Cultivo Primario de Células , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Receptor Toll-Like 4/inmunología , Factor de Necrosis Tumoral alfa/metabolismo
12.
Inflammation ; 37(2): 478-85, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24202549

RESUMEN

Mastitis is characterized by an inflammation of the mammary gland of dairy animals and humans; this condition is one of the major causes of economic losses in dairy industries. Selenium (Se), a biological trace element, modulates the functions of many regulatory proteins in signal transduction and provides advantages for animals with inflammatory diseases, including mastitis. The current study aimed to assess the protective effects and the active mechanism of Na(2)SeO(3) against lipopolysaccharide (LPS)-induced inflammation in mouse mammary epithelial cells (MMECs). Our results showed that LPS-induced expressions of cyclooxygenase-2 and tumor necrosis factor-α significantly decreased after Se was supplemented to Se-deficient MMECs. Na(2)SeO(3) also suppressed LPS-induced nuclear factor-κB activation, inhibitory kappa B degradation, and ERK, JNK, and P38 phosphorylation in a dose-dependent manner. These results suggested that Se functions as an anti-inflammatory agent in mastitis.


Asunto(s)
Antiinflamatorios/farmacología , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/farmacología , Glándulas Mamarias Animales/efectos de los fármacos , Mastitis/prevención & control , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Selenito de Sodio/farmacología , Animales , Células Cultivadas , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Relación Dosis-Respuesta a Droga , Activación Enzimática , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas I-kappa B/metabolismo , Glándulas Mamarias Animales/enzimología , Glándulas Mamarias Animales/inmunología , Mastitis/enzimología , Mastitis/genética , Mastitis/inmunología , Ratones , Fosforilación , Embarazo , Cultivo Primario de Células , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo
13.
J Mammary Gland Biol Neoplasia ; 16(4): 305-22, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21968536

RESUMEN

Application of microarrays to the study of intramammary infections in recent years has provided a wealth of fundamental information on the transcriptomics adaptation of tissue/cells to the disease. Due to its heavy toll on productivity and health of the animal, in vivo and in vitro transcriptomics works involving different mastitis-causing pathogens have been conducted on the mammary gland, primarily on livestock species such as cow and sheep, with few studies in non-ruminants. However, the response to an infectious challenge originating in the mammary gland elicits systemic responses in the animal and encompasses tissues such as liver and immune cells in the circulation, with also potential effects on other tissues such as adipose. The susceptibility of the animal to develop mastitis likely is affected by factors beyond the mammary gland, e.g. negative energy balance as it occurs around parturition. Objectives of this review are to discuss the use of systems biology concepts for the holistic study of animal responses to intramammary infection; providing an update of recent work using transcriptomics to study mammary and peripheral tissue (i.e. liver) as well as neutrophils and macrophage responses to mastitis-causing pathogens; discuss the effect of negative energy balance on mastitis predisposition; and analyze the bovine and murine mammary innate-immune responses during lactation and involution using a novel functional analysis approach to uncover potential predisposing factors to mastitis throughout an animal's productive life.


Asunto(s)
Adaptación Fisiológica , Infecciones Bacterianas/genética , Infecciones Bacterianas/veterinaria , Glándulas Mamarias Humanas/fisiología , Mastitis/genética , Mastitis/inmunología , Transcriptoma , Animales , Infecciones Bacterianas/inmunología , Bovinos , Femenino , Humanos , Inmunidad Innata , Glándulas Mamarias Humanas/microbiología , Mastitis/microbiología
14.
Clin Vaccine Immunol ; 18(6): 893-900, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21508165

RESUMEN

To construct a universal vaccine against mastitis induced by either Streptococcus agalactiae or Staphylococcus aureus, the B cell epitopes of the surface immunogenic protein (Sip) from S. agalactiae and clumping factor A (ClfA) from S. aureus were analyzed and predicted. sip-clfA, a novel chimeric B cell epitope-based gene, was obtained by overlap PCR, and then the recombinant Sip-ClfA (rSip-ClfA) was expressed and purified. rSip-ClfA and inactivated S. agalactiae and S. aureus were formulated into different vaccines with mineral oil as the adjuvant and evaluated in mouse models. The rSip-ClfA vaccination induced immunoglobulin G (IgG) titers higher than those seen in groups immunized with inactivated bacteria. Furthermore, the response to rSip-ClfA immunization was characterized as having a dominant IgG1 subtype, whereas both bacterial immunizations produced similar levels of IgG1 and IgG2a. The antiserum capacities for opsonizing adhesion and phagocytosis were significantly greater in the rSip-ClfA immunization group than in the killed-bacterium immunization groups (P < 0.05). The immunized lactating mice were challenged with either S. agalactiae or S. aureus via the intramammary route. At 24 h postinfection, the numbers of bacteria recovered from the mammary glands in the rSip-ClfA group were >5-fold lower than those in both inactivated-bacterium groups (P < 0.01). Histopathological examination of the mammary glands showed that rSip-ClfA immunization provided better protection of mammary gland tissue integrity against both S. agalactiae and S. aureus challenges. Thus, the recombinant protein rSip-ClfA would be a promising vaccine candidate against mastitis induced by either S. agalactiae or S. aureus.


Asunto(s)
Epítopos de Linfocito B/inmunología , Mastitis/prevención & control , Vacunas Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Streptococcus agalactiae/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Antibacterianos/sangre , Carga Bacteriana , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Bovinos , Coagulasa/genética , Coagulasa/inmunología , Epítopos de Linfocito B/genética , Femenino , Histocitoquímica , Inmunoglobulina G/sangre , Glándulas Mamarias Animales/microbiología , Mastitis/inmunología , Mastitis/microbiología , Ratones , Ratones Endogámicos BALB C , Aceite Mineral/administración & dosificación , Proteínas Opsoninas/sangre , Fagocitosis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Vacunas Estafilocócicas/administración & dosificación , Vacunas Estafilocócicas/genética , Staphylococcus aureus/genética , Staphylococcus aureus/aislamiento & purificación , Streptococcus agalactiae/genética , Streptococcus agalactiae/aislamiento & purificación , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología
15.
Clin Vaccine Immunol ; 17(11): 1746-52, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20826613

RESUMEN

The present study evaluated the potential of recombinant binding region A of clumping factor A (rClfA-A) to be an effective component of a vaccine against mastitis induced by Staphylococcus aureus in the mouse. rClfA-A and inactivated S. aureus were each emulsified in Freund's adjuvant, mineral oil adjuvant, and Seppic adjuvant; phosphate-buffered saline was used as a control. Seven groups of 12 mice each were immunized intraperitoneally three times at 2-week intervals. The titers of IgG and subtypes thereof (IgG1 and IgG2a) in the rClfA-A-immunized group were more than 1,000-fold higher than those in the killed-bacteria-immunized group (P < 0.01). Of the three adjuvants used, mineral oil adjuvant induced the highest antibody levels for both antigens (P < 0.001). Furthermore, the anti-rClfA-A antibody capacities for bacterial adhesion and opsonizing phagocytosis were significantly greater in the rClfA-A-immunized group than in the killed-bacteria-immunized group (P < 0.05). Lactating mice immunized with either rClfA-A or inactivated vaccine were challenged with S. aureus via the intramammary route. The numbers of bacteria recovered from the murine mammary glands 24 h after inoculation were significantly lower in the rClfA-A group than in the killed-bacteria-immunized group (P < 0.001). Histologic examination of the mammary glands showed that rClfA-A immunization effectively preserved tissue integrity. Thus, rClfA-A emulsified in an oil adjuvant provides strong immune protection against S. aureus-induced mastitis in the mouse.


Asunto(s)
Coagulasa/inmunología , Mastitis/prevención & control , Vacunas Estafilocócicas/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Antibacterianos/sangre , Adhesión Bacteriana/inmunología , Carga Bacteriana , Coagulasa/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Adyuvante de Freund/administración & dosificación , Histocitoquímica , Inmunoglobulina G/sangre , Inyecciones Intraperitoneales , Masculino , Glándulas Mamarias Animales/microbiología , Glándulas Mamarias Animales/patología , Mastitis/inmunología , Ratones , Ratones Endogámicos BALB C , Microscopía , Proteínas Opsoninas/sangre , Fagocitosis/inmunología , Vacunas Estafilocócicas/administración & dosificación , Vacunas de Subunidad/inmunología
16.
Vaccine ; 28(24): 4038-44, 2010 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-20416265

RESUMEN

The immunoprotective effect of the ligand-binding domain of fibronectin-binding protein (lFnBP) from Staphylococcus aureus (S. aureus) was investigated in a mouse mastitis model. The recombinant lFnBP (rlFnBP) and inactivated S. aureus were emulsified in Freund's adjuvant, mineral oil adjuvant or Seppic adjuvant, respectively. Seven groups of mice (n=12 each) were immunized with these six vaccines or phosphate-buffered saline. The immunoglobulin G (IgG) titers of mice immunized with rlFnBP vaccine were higher than those in the inactivated vaccine group (P<0.01). Antiserum capacities to opsonize adhesion and phagocytosis were significantly greater in the rlFnBP immunization group than in the killed bacteria group (P<0.05). The immunized lactating mice were challenged with S. aureus. At 24h postinfection, the numbers of bacteria recovered in the rlFnBP group were significantly lower than those in the killed bacteria group (P<0.001). Levels of both interleukin-6 (IL-6) and interferon-gamma (IFN-gamma from the mammary glands in the rlFnBP group were higher than those in the inactivated group (P<0.05). Better protection of mammary gland tissue was shown in the rlFnBP group. Thus, the rlFnBP, emulsified in an oil adjuvant, provided strong immune protection against S. aureus mastitis in mice, and could therefore be a promising vaccine candidate against bovine mastitis induced by S. aureus.


Asunto(s)
Adhesinas Bacterianas/inmunología , Mastitis/inmunología , Infecciones Estafilocócicas/inmunología , Vacunas Estafilocócicas/inmunología , Adyuvantes Inmunológicos/farmacología , Animales , Anticuerpos Antibacterianos/sangre , Formación de Anticuerpos , Bovinos , Femenino , Inmunoglobulina G/sangre , Interferón gamma/inmunología , Interleucina-6/inmunología , Glándulas Mamarias Animales/microbiología , Glándulas Mamarias Animales/patología , Mastitis/prevención & control , Ratones , Ratones Endogámicos BALB C , Neutrófilos/inmunología , Fagocitosis , Proteínas Recombinantes/inmunología , Infecciones Estafilocócicas/prevención & control
17.
Vet Res Commun ; 32(4): 305-13, 2008 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-18163219

RESUMEN

Antioxidant, antiinflammatory and phagocytic activities were studied in milk polymorphonuclear cells (PMNs) isolated from healthy buffaloes (group I) and during clinical mastitis with the treatment of Enrofloxacin alone (group II) and combined treatment with Enrofloxacin and Vitamin E plus selenium (group III). On days 0,3, 8 and 15 the milk Somatic cell count (SCC) were significantly higher in mastitic milk than in milk obtained from healthy buffaloes. In group II SCC decreased significantly on day 3 and day 8, however in group III reduction in SCC was observed on day 3, day 8 and day 15 (P < 0.05). The antiinflammatory activity was evaluated by determining nitrite plus nitrate (NOx) production in the milk PMNs before treatment and on day 8. NOx activity was significantly higher in mastitic milk than from healthy controls, both before and after treatment (P < 0.05). In group II and group III the activity decreased significantly on day 8 (P < 0.05). The Glutathione peroxidase (GSH-Px) activity was estimated in the milk polymorphonuclear cell (PMNs) supernatant. GSH-Px activity was significantly lower in mastitic buffaloes than in healthy controls, both before and after treatment (P < 0.05). In group II levels did not change in response to treatment, whereas in group III levels had increased significantly on day 8 (P < 0.05). The phagocytic activity (PA) (percentage of neutrophil that had phagocytosed 1-6 bacteria) and phagocytic index (PI) (average number of bacteria/ leukocytes counted in 100 cells) of the milk PMNs was significantly lower in mastitic buffaloes (P < 0.05). In group II the PA and PI did not change in response to treatment, whereas in group III both the parameters had increased significantly on day 8 (P < 0.05). The results of the present experiment indicated enhancement of antioxidative and cellular defense and reduction of somatic cell count in the mastitic animals treated with Enrofloxacin and Vitamin E plus Selenium as compared to the Enrofloxacin treatment alone. Hence Vitamin E plus selenium therapy may be added along with the antibiotics for effective amelioration of intramammary infection in buffaloes.


Asunto(s)
Antibacterianos/farmacología , Búfalos/microbiología , Fluoroquinolonas/farmacología , Mastitis/veterinaria , Neutrófilos/efectos de los fármacos , Selenio/farmacología , Vitamina E/farmacología , Animales , Antioxidantes/farmacología , Búfalos/inmunología , Búfalos/metabolismo , Recuento de Células/veterinaria , Enrofloxacina , Femenino , Glutatión Peroxidasa/metabolismo , Mastitis/tratamiento farmacológico , Mastitis/inmunología , Mastitis/metabolismo , Mastitis/microbiología , Leche/citología , Leche/inmunología , Leche/microbiología , Neutrófilos/inmunología , Óxido Nítrico/metabolismo , Fagocitosis/efectos de los fármacos
18.
Am J Clin Nutr ; 84(5): 1086-92, 2006 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17093161

RESUMEN

BACKGROUND: Multiple micronutrient supplementation of Nepalese women during pregnancy is associated with a significant increase in birth weight. OBJECTIVE: We tested the hypothesis that improved birth weight in infants of mothers supplemented with micronutrients is associated with a decrease in inflammatory responses and an increase in the production of T helper 1 cells and T helper 2 cells. DESIGN: The study was embedded in a randomized controlled trial of 15 micronutrients, compared with iron-folate supplementation (control), given during pregnancy with the aim of increasing birth weight. Blood samples were collected at 32 wk of gestation, 12-20 wk after supplementation began, for the measurement of inflammatory markers. Breast-milk samples were collected 1 mo after delivery for the measurement of the ratio of milk sodium to potassium (milk Na:K). In an opportunistically selected subgroup of 70 women, mitogen-stimulated cytokine production was measured ex vivo in whole blood. RESULTS: Blood eosinophils; plasma concentrations of the acute phase reactants C-reactive protein, alpha(1)-acid glycoprotein (AGP), neopterin, and ferritin; milk Na:K; and the production of interleukin (IL) 10, IL-4, interferon gamma, and tumor necrosis factor alpha in whole blood did not differ significantly between the supplemented and control groups. Plasma C-reactive protein and AGP were higher in women who had a preterm delivery, and AGP was higher in women who delivered a low-birth-weight term infant than in women who delivered a normal-birth-weight term infant. CONCLUSIONS: The results indicate an association between systemic inflammation in late pregnancy and compromised delivery outcome in Nepalese women but do not support the hypothesis that multiple micronutrient supplementation changes cytokine production or inflammatory markers.


Asunto(s)
Peso al Nacer/efectos de los fármacos , Proteína C-Reactiva/análisis , Inflamación/sangre , Micronutrientes/administración & dosificación , Orosomucoide/análisis , Embarazo/inmunología , Adulto , Proteína C-Reactiva/metabolismo , Suplementos Dietéticos , Femenino , Humanos , Recién Nacido de Bajo Peso , Recién Nacido , Interferón gamma/biosíntesis , Interleucina-10/biosíntesis , Interleucina-4/biosíntesis , Mastitis/inmunología , Leche Humana/química , Nepal , Orosomucoide/metabolismo , Potasio/análisis , Potasio/metabolismo , Embarazo/sangre , Resultado del Embarazo , Atención Prenatal/métodos , Sodio/análisis , Sodio/metabolismo , Células TH1/inmunología , Células Th2/inmunología
19.
J Pineal Res ; 41(2): 136-41, 2006 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-16879319

RESUMEN

A large number of data show that melatonin has immunomodulatory properties and is produced by immunocompetent cells; also, some evidence suggests a 'feedback' of the activated immune system on the pineal gland. In this paper, we studied immune-pineal interactions in colostrum obtained from healthy puerperae and mothers with mastitis taking into account that, (a) melatonin levels in milk reflects pineal activity and (b) colostrum quiescent mononuclear and polymorphonuclear phagocytes from healthy mothers in culture are adequate for evaluating the ability of immunocompetent cells to produce melatonin. Here we compared the diurnal and nocturnal melatonin levels in colostrum from healthy puerperae and mothers with mastitis; this is a unique noninvasive model for determining pineal activity in the proinflammatory phase of a defense response. In addition, we determined the 'in vitro' production of melatonin by colostrum immunocompetent cells stimulated by enteropathogenic Escherichia coli or zymosan. Suppression of nocturnal melatonin rise in mothers with mastitis was highly correlated with increased tumor necrosis factor-alpha (TNF-alpha) secretion. This result, interpreted taking into account the presence of the transcription factor nuclear factor kappa B in pineal gland, suggest that the proinflammatory cytokine can inhibit nocturnal pineal melatonin production. On the other hand, stimulated, but not quiescent, immunocompetent cells secreted in the colostrum produced melatonin in vitro. In addition, this production ceases after bacteria killing. These results suggest that during the response to an injury the production of melatonin can be transiently shifted from an endocrine (pineal) to a paracrine (immunocompetent cells) source.


Asunto(s)
Calostro/química , Mastitis/metabolismo , Melatonina/biosíntesis , Comunicación Paracrina , Fagocitos/metabolismo , Glándula Pineal/metabolismo , Adolescente , Adulto , Calostro/inmunología , Escherichia coli/inmunología , Femenino , Humanos , Lactante , Mastitis/inmunología , Melatonina/análisis , Fagocitos/inmunología , Periodo Posparto , Embarazo , Factor de Necrosis Tumoral alfa/análisis , Zimosan/inmunología
20.
Vet Res Commun ; 26(8): 587-600, 2002 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-12507034

RESUMEN

A double-blind, placebo-controlled study was conducted to compare two vaccines using different adjuvants with regard to their ability to stimulate antibody production against the alpha- and beta-toxins and the exopolysaccharide of Staphylococcus aureus. The vaccines contained identical antigens, consisting of inactivated whole bacteria of two strains of S. aureus in addition to alpha- and beta-toxoid. One vaccine contained mineral oil, while the other used a water-soluble acrylic acid polymer resin (Carbopol) as adjuvant. Saline served as the placebo. One hundred and forty ewes were vaccinated twice before lambing, by subcutaneous injection with vaccine or placebo in the region of the supramammary lymph node, and were observed and sampled over a period of 6 months. The vaccine containing mineral oil as adjuvant induced significantly greater immune responses to the alpha- and beta-toxins than did the vaccine containing Carbopol. The latter vaccine induced higher levels of antibodies to exopolysaccharide. The degree of local adverse reactions did not differ between the two groups. The results indicate differences between the oil-adjuvanted and Carbopol-adjuvanted vaccines with regard to their ability to stimulate antibody production against S. aureus protein antigens in sheep.


Asunto(s)
Adyuvantes Inmunológicos/farmacología , Anticuerpos Antibacterianos/biosíntesis , Inmunoglobulina G/biosíntesis , Mastitis/veterinaria , Enfermedades de las Ovejas/inmunología , Infecciones Estafilocócicas/veterinaria , Vacunas Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Resinas Acrílicas , Animales , Anticuerpos Antibacterianos/sangre , Método Doble Ciego , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Inmunoglobulina G/sangre , Mastitis/inmunología , Mastitis/microbiología , Mastitis/prevención & control , Leche/microbiología , Aceite Mineral/farmacología , Polivinilos/farmacología , Embarazo , Ovinos , Enfermedades de las Ovejas/microbiología , Infecciones Estafilocócicas/sangre , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/microbiología , Vacunas de Productos Inactivados/inmunología , Vacunas de Productos Inactivados/normas
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