RESUMEN

As an essential trace element for animals, copper significantly contributes to the growth and health of animals. Compared to inorganic trace elements, organic trace elements are better supplements; notably, they are acquired through microbial transformation. Therefore, we screened for copper-enriched microorganisms from high copper content soil to obtain organic copper. Sodium diethyldithio carbamate trihydrate was applied as a chromogenic agent for determining micro amounts of intracellular copper through spectrophotometry. In total, 50 fungi were isolated after the successful application of the screening platform for copper-rich microbes. Following morphological and molecular biology analyses, the N-2 strain, identified as Aspergillus niger sp. demonstrated showed better copper enrichment potential than others. Notably, the strain tolerance to copper was nearly thrice that of Saccharomyces cerevisiae, up to 1600mg/L. The content of the organic bound copper was 22.84mg Cu/g dry cell. Using the Central Composite Design (CCD) response surface method, we optimized the fermentation condition (inoculation amount, 13%; temperature, 28(C; pH, 5.0). Compared to the original strain results under the single factor fermentation condition, we reported an increase by 24.18% under the optimized conditions. Collectively, these findings provide a reference for uncovering new and low-cost organic copper additives.(AU)

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Como elemento traço essencial para os animais, o cobre contribui significativamente para o crescimento e saúde dos animais. Comparado aos oligoelementos inorgânicos, os oligoelementos orgânicos são melhores suplementos; notavelmente, eles são adquiridos através de transformação microbiana. Portanto, nós selecionamos microorganismos enriquecidos com cobre de solos com alto teor de cobre para obter cobre orgânico. O carbamato de sódio diethyldithio trihidratado foi aplicado como agente cromogênico para a determinação de micro quantidades de cobre intracelular através da espectrofotometria. No total, 50 fungos foram isolados após a aplicação bem sucedida da plataforma de triagem para micróbios ricos em cobre. Após análises morfológicas e de biologia molecular, a cepa N-2, identificada como Aspergillus niger sp. demonstrou um melhor potencial de enriquecimento de cobre do que outras. Notavelmente, a tolerância da estirpe ao cobre foi quase três vezes maior que a da Saccharomyces cerevisiae, até 1600mg/L. O conteúdo de cobre ligado orgânico era de 22,84mg Cu/g de célula seca. Usando o método de superfície de resposta Central Composite Design (CCD), nós otimizamos a condição de fermentação (quantidade de inoculação, 13%; temperatura, 28C; pH, 5,0). Em comparação com os resultados da deformação original sob a condição de fermentação de fator único, relatamos um aumento de 24,18% sob as condições otimizadas. Coletivamente, estas descobertas fornecem uma referência para descobrir novos aditivos de cobre orgânico de baixo custo.(AU)

Asunto(s)

Animales , Análisis del Suelo , Cobre , Aditivos Alimentarios , Aspergillus , Microbiología del Suelo , Tratamiento del Suelo , Sus scrofa

RESUMEN

We investigated the egg production, changes in luteinizing hormone (LH), follicle-stimulating hormone (FSH), gonadal hormones, and their mRNA levels in the hypothalamic-pituitary-gonadal axis of White King pigeons submitted to different photoperiods. The treatments consisted of three photoperiods (8 h light (L):16 h dark (D), 12L:12D, and 16L:8D), with three replicates of twelve pairs of adult pigeons. The birds were exposed the photoperiods for 45 days. Egg production performance was recorded daily. Six pigeon pairs per replicate were selected for plasma collection, and six pigeon pairs per replicate for the resection of the hypothalamic-pituitary-gonadal (HPG) axis. Egg production was significantly improved by long-day lighting (16L:8D), while no differences in egg shape index were detected. Higher average egg weight was obtained in 16L:8D group, whereas broken egg percentage was higher in the 8L:16D group. Female LH level was significantly higher in long-day lighting, and the FSH level significantly lower in short-day lighting. The females in the 16L:8D group had higher estrogen level. The photoperiods had a minor effect on plasma LH and testosterone in males, whereas the FSH level was significantly higher in the 16L:8D group. The level of LH mRNA expression was higher in both females and males of the 16L:8D group. Similar trends in FSH mRNA expression observed in both females and males. The 16L:8D photoperiod not only improved egg production, but also stimulated plasma LH, FSH, gonadal hormones, and promoted LH and FSH mRNA expression in pigeons.

Asunto(s)

Animales , Columbidae/fisiología , Hormona Folículo Estimulante/análisis , Hormona Luteinizante/administración & dosificación , Hormona Luteinizante/análisis , Hormonas Gonadales/análisis , Hormona Liberadora de Gonadotropina/análisis

RESUMEN

We investigated the egg production, changes in luteinizing hormone (LH), follicle-stimulating hormone (FSH), gonadal hormones, and their mRNA levels in the hypothalamic-pituitary-gonadal axis of White King pigeons submitted to different photoperiods. The treatments consisted of three photoperiods (8 h light (L):16 h dark (D), 12L:12D, and 16L:8D), with three replicates of twelve pairs of adult pigeons. The birds were exposed the photoperiods for 45 days. Egg production performance was recorded daily. Six pigeon pairs per replicate were selected for plasma collection, and six pigeon pairs per replicate for the resection of the hypothalamic-pituitary-gonadal (HPG) axis. Egg production was significantly improved by long-day lighting (16L:8D), while no differences in egg shape index were detected. Higher average egg weight was obtained in 16L:8D group, whereas broken egg percentage was higher in the 8L:16D group. Female LH level was significantly higher in long-day lighting, and the FSH level significantly lower in short-day lighting. The females in the 16L:8D group had higher estrogen level. The photoperiods had a minor effect on plasma LH and testosterone in males, whereas the FSH level was significantly higher in the 16L:8D group. The level of LH mRNA expression was higher in both females and males of the 16L:8D group. Similar trends in FSH mRNA expression observed in both females and males. The 16L:8D photoperiod not only improved egg production, but also stimulated plasma LH, FSH, gonadal hormones, and promoted LH and FSH mRNA expression in pigeons.(AU)

Asunto(s)

Animales , Columbidae/fisiología , Hormona Luteinizante/administración & dosificación , Hormona Luteinizante/análisis , Hormona Folículo Estimulante/análisis , Hormonas Gonadales/análisis , Hormona Liberadora de Gonadotropina/análisis

RESUMEN

A dose-response experiment with four dietary copper concentrations (4.17, 8.17, 12.17 and 16.17 mg/kg) was conducted to estimate the growth performance, slaughter performance, nutrient content of fecal and liver copper concentrations of growing Goslings from 28 to 70 d of age. Two hundred healthy male Yangzhou geese with similar body weight were randomized to four groups with five replicates per treatment and ten geese per replicate. Average daily feed intake, average daily gain and feed conversion ratio of geese for each pen were measured from 28 to 70 d of age. At 70 d of age, two geese were selected randomly from each pen and slaughtered to evaluate carcass quality. Metabolism experiment was conducted with five male geese from each group (one goose per pen) which body weight was close to the mean weight of the group from 64 to 70 d of age. Significant effects of dietary copper was found on body weight, feed conversion ratio, carcass yield, fecal copper concentrations and liver copper concentrations. Body weight, feed conversion ratio and carcass yield showed significant quadratic response to increase dietary copper concentration, while fecal copper concentration and liver copper concentration showed a significant linear response. The result showed that dietary Cu addition can improve growth by increasing the use of the feeding stuff and improving carcass yield in growing Goslings. Furthermore, taking into consideration, the optimal level of Gosling dietary copper was between 8.77 and 11.6 mg/kg from 28 to 70 days of age.

Asunto(s)

Animales , Recién Nacido , Sacrificio de Animales , Cobre/análisis , Gansos/anomalías , Gansos/fisiología , Heces/química

RESUMEN

A dose-response experiment with four dietary copper concentrations (4.17, 8.17, 12.17 and 16.17 mg/kg) was conducted to estimate the growth performance, slaughter performance, nutrient content of fecal and liver copper concentrations of growing Goslings from 28 to 70 d of age. Two hundred healthy male Yangzhou geese with similar body weight were randomized to four groups with five replicates per treatment and ten geese per replicate. Average daily feed intake, average daily gain and feed conversion ratio of geese for each pen were measured from 28 to 70 d of age. At 70 d of age, two geese were selected randomly from each pen and slaughtered to evaluate carcass quality. Metabolism experiment was conducted with five male geese from each group (one goose per pen) which body weight was close to the mean weight of the group from 64 to 70 d of age. Significant effects of dietary copper was found on body weight, feed conversion ratio, carcass yield, fecal copper concentrations and liver copper concentrations. Body weight, feed conversion ratio and carcass yield showed significant quadratic response to increase dietary copper concentration, while fecal copper concentration and liver copper concentration showed a significant linear response. The result showed that dietary Cu addition can improve growth by increasing the use of the feeding stuff and improving carcass yield in growing Goslings. Furthermore, taking into consideration, the optimal level of Gosling dietary copper was between 8.77 and 11.6 mg/kg from 28 to 70 days of age.(AU)

Asunto(s)

Animales , Recién Nacido , Cobre/análisis , Sacrificio de Animales , Gansos/anomalías , Gansos/fisiología , Heces/química

RESUMEN

Genomic imprinting is an important epigenetic mechanism that has vital effects on fetal growth and development. We observed the differences in four tissues (heart, spleen, liver, and kidney) from dead transgenic cloned goats using hematoxylin and eosin (H&E) staining. Eight imprinted genes in the tissues of dead transgenic cloned and normal goats were analyzed using reverse transcription polymerase chain reaction. H&E staining results from the abortion group indicated the lack of obvious morphological changes in heart and spleen tissues, while inflammatory cell infiltration and glomerular nephritis characteristics were observed in liver and kidney tissues, respectively. Compared to the control group, CDKN1C, H19, IGF2R, and SNRPN were significantly (P < 0.05) overexpressed in the heart tissue of the abortion group, while XIST was significantly reduced. In the liver tissues, CDKN1C and DLK1 expression decreased, while GNAS, H19, IGF2R, PEG3, and XIST expression increased significantly. In the spleen tissues, DLK1 expression increased, while GNAS, H19, IGF2R, PEG3, SNRPN, and XIST expression decreased. In the kidney tissues, CDKN1C, DLK1, GNAS, IGF2R, and PEG3 expression increased, while H19 and XIST expression decreased. The overall expression of imprinted genes was abnormal in different tissues of transgenic cloned goats, and the degree of abnormal genomic imprinting was more severe in the abortion group compared to the death and control groups. These results suggest that abnormal expression of imprinted genes may cause developmental defects in transgenic cloned goats. Moreover, abnormal epigenetic modifications may affect the reprogramming of transgenic donor cells.

Asunto(s)

Clonación de Organismos/mortalidad , Epigénesis Genética , Genes Letales , Impresión Genómica , Cabras/genética , Lactoferrina/genética , Animales , Animales Modificados Genéticamente , Inhibidor p57 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p57 de las Quinasas Dependientes de la Ciclina/metabolismo , Femenino , Perfilación de la Expresión Génica , Cabras/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Riñón/metabolismo , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Lactoferrina/metabolismo , Hígado/metabolismo , Masculino , Miocardio/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Embarazo , Transducción de Señal , Bazo/metabolismo , Transgenes

RESUMEN

Resistin (RSTN) expression in subcutaneous adipose tissue, and its effect on glucose metabolism in rats with traumatic brain injury, was investigated using real-time PCR, western blots, and enzyme linked immunoassays. Our results show that the expression of RSTN mRNA (3.192 ± 0.046, 4.016 ± 0.010, 6.004 ± 0.020, 8.213 ± 0.013, 11.199 ± 0.174, 15.094 ± 0.030), protein levels (1.79 ± 0.05, 1.98 ± 0.07, 2.75 ± 0.08, 3.19 ± 0.08, 4.25 ± 0.11, 4.48 ± 0.07), levels of serum insulin (512.96 ± 1.21, 580.57 ± 1.52, 769.71 ± 2.22, 826.08 ± 2.03, 1262.25 ± 3.40, 1512.80 ± 3.93), and fasting blood glucose levels (10.277 ± 0.040, 12.776 ± 0.038, 13.403 ± 0.263, 14.698 ± 0.100, 16.637 ± 0.110, 19.416 ± 0.025) were significantly higher in the traumatic rat group compared to the control group (P < 0. 05). Quantitative insulin sensitivity check index (QUICKI) was significantly lower in the traumatic group (-8.570 ± 0.005, -8.912 ± 0.004, -9.241 ± 0.022, -9.404 ± 0.007, -9.952 ± 0.007, -10.288 ± 0.002) than in the control group (-7.633 ± 0.003, -7.639 ± 0.004, -7.637 ± 0.006, -7.643 ± 0.003, -7.636 ± 0.006, -7.634 ± 0.004) (P < 0.05). Single factor linear correlation analysis showed that there was a significant negative correlation between RSTN expression and QUICKI (-0.983, P < 0.05) in the traumatic group. The increase in RSTN expression in the subcutaneous adipose tissue of rats with traumatic brain injury is likely related to the indexes of glycometabolism, including serum insulin, fasting blood glucose, and QUICKI. Our results lead us to conclude that RSTN may play an important role in the process of insulin resistance in rats with traumatic brain injury.

Asunto(s)

Glucemia/metabolismo , Lesiones Encefálicas/genética , ARN Mensajero/genética , Resistina/genética , Grasa Subcutánea/metabolismo , Animales , Lesiones Encefálicas/sangre , Lesiones Encefálicas/patología , Metabolismo de los Hidratos de Carbono/genética , Ayuno , Expresión Génica , Insulina/sangre , Resistencia a la Insulina , Masculino , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Resistina/metabolismo , Grasa Subcutánea/patología

RESUMEN

Cells isolated from human first trimester umbilical cord perivascular layer (hFTM-PV) tissues display the pluripotent characteristics of stem cells. In this study, we examined whether hFTM-PV cells can differentiate into islet-like clusters (ILCs) in vitro, and whether transplantation of the hFTM-PV cells with and without differentiation in vitro can alleviate diabetes in nude mice. The hFTM-PV cells were differentiated into ILCs in vitro through a simple stepwise culture protocol. To examine the in vivo effects of the cells, the hFTM-PV cells with and without differentiation in vitro were transplanted into the abdominal cavity of nude mice with streptozotocin (STZ)-induced diabetes. Blood glucose levels, body weight, and the survival probability of the diabetic nude mice were then statistically analyzed. The hFTM-PV cells were successfully induced into ILCs that could release insulin in response to elevated concentrations of glucose in vitro. In transplantation experiments, we observed that mice transplanted with the undifferentiated hFTM-PV cells, embryonic body-like cell aggregations, or ILCs all demonstrated normalized hyperglycemia and showed improved survival rate compared with those without cell transplantation. The hFTM-PV cells have the ability to differentiate into ILCs in vitro and transplantations of undifferentiated and differentiated cells can alleviate STZ-induced diabetes in nude mice. This may offer a potential cell source for stem cell-based therapy for treating diabetes in the future.

Asunto(s)

Trasplante de Células Madre de Sangre del Cordón Umbilical/métodos , Diabetes Mellitus Experimental/terapia , Animales , Diferenciación Celular/fisiología , Diabetes Mellitus Experimental/sangre , Femenino , Humanos , Islotes Pancreáticos/citología , Ratones , Ratones Desnudos , Embarazo , Cordón Umbilical/citología

RESUMEN

The large yellow croaker (Larimichthys crocea) is one of the largest marine net-cage cultured species in the oceans around China. In the present study, we isolated and characterized 13 polymorphic microsatellite markers from genomic libraries of L. crocea. Loci were screened for 10 wild specimens from 2 sites in southeast of China. All loci were polymorphic. The number of alleles per locus ranged from 2 to 21. The expected heterozygosity ranged from 0.233 to 0.838 and observed heterozygosity ranged from 0.527 to 0.935. Eleven loci were highly informative (polymorphic information content >0.5). Significant deviation from Hardy-Weinberg equilibrium was observed at 3 loci after Bonferroni's correction. The microsatellite loci may be valuable tools for studying the genetic diversity and genetic structure for conservation planning of the fish.

Asunto(s)

Sitios Genéticos , Repeticiones de Microsatélite , Perciformes/genética , Polimorfismo Genético , Animales

RESUMEN

We investigated the effects of a modified Shoutaiwai recipe on integrin ß3 and leukemia-inhibitory factor (LIF) in the endometrium of controlled ovarian hyperstimulation (COH) mice during the implantation window. Seventy non-pregnant mice were randomly divided into 3 groups: a traditional medicine (TCM) treatment group (N = 30), an aspirin treatment (N = 30) group, and a control group (N = 10). After the model was successfully established, mice in the drug treatment groups and the control group were respectively treated with the modified Shoutaiwai recipe, aspirin, or 0.9% physiological saline. During the implantation window of mice, the middle segment of the mouse uterus was recovered, and integrin ß3 and LIF expressions in the endometrium were respectively detected using an immunohistological two-step method and reverse transcription-PCR. Expressions of integrin ß3 and LIF in the endometrium of mice in the TCM treatment group were significantly increased compared to aspirin-treated and control mice, and those of aspirin-treated mice were increased compared to the control group. Our modified Shoutaiwai recipe may improve the endometrial receptivity of COH mice by increasing the expression of integrin ß3 and LIF in the endometrium during the implantation window.

Asunto(s)

Dieta/veterinaria , Implantación del Embrión/fisiología , Endometrio/metabolismo , Integrina beta3/metabolismo , Factor Inhibidor de Leucemia/metabolismo , Inducción de la Ovulación/métodos , Animales , Aspirina/farmacología , Dietoterapia , Medicamentos Herbarios Chinos/farmacología , Endometrio/patología , Femenino , Ratones , Modelos Animales , Embarazo

RESUMEN

Axonopus compressus (Sw.) Beauv. is a perennial herb widely used as a garden lawn grass. In this study, we used Roche 454 pyrosequencing, combined with the magnetic bead enrichment method FIASCO, to isolate simple sequence repeat markers from the A. compressus genome. A total of 1942 microsatellite loci were identified, with 53,193 raw sequencing reads. One hundred micro-satellite loci were selected to test the primer amplification efficiency in 24 individuals; 14 primer pairs yielded polymorphic amplification products. The number of observed alleles ranged from two to six, with an average of 3.5. Shannon's Information index values ranged from 0.169 to 0.650, with an average of 0.393. Nei's genetic diversity values ranged from 0.108 to 0.457, with an average of 0.271. This first set of microsatellite markers developed for Axonopus will assist in the development of molecular marker-assisted breeding and the assessment of genetic diversity in A. compressus.

Asunto(s)

Marcadores Genéticos , Repeticiones de Microsatélite/genética , Poaceae/genética , Análisis de Secuencia de ADN/métodos , Filogenia , Poaceae/clasificación

RESUMEN

The present study focuses on the neuroprotective effect of glycyrrhizic acid (GA, a major compound separated from Glycyrrhiza Radix, which is a crude Chinese traditional drug) against glutamate-induced cytotoxicity in differentiated PC12 (DPC12) cells. The results showed that GA treatment improved cell viability and ameliorated abnormal glutamate-induced alterations in mitochondria in DPC12 cells. GA reversed glutamate-suppressed B-cell lymphoma 2 levels, inhibited glutamate-enhanced expressions of Bax and cleaved caspase 3, and reduced cytochrome C (Cyto C) release. Exposure to glutamate strongly inhibited phosphorylation of AKT (protein kinase B) and extracellular signal-regulated kinases (ERKs); however, GA pretreatment enhanced activation of ERKs but not AKT. The presence of PD98059 (a mitogen-activated protein/extracellular signal-regulated kinase kinase [MEK] inhibitor) but not LY294002 (a phosphoinositide 3-kinase [PI3K] inhibitor) diminished the potency of GA for improving viability of glutamate-exposed DPC12 cells. These results indicated that ERKs and mitochondria-related pathways are essential for the neuroprotective effect of GA against glutamate-induced toxicity in DPC12 cells. The present study provides experimental evidence supporting GA as a potential therapeutic agent for use in the treatment of neurodegenerative diseases.

Asunto(s)

Animales , Ratas , Antiinflamatorios/uso terapéutico , Ácido Glutámico/toxicidad , Ácido Glicirrínico/uso terapéutico , Fármacos Neuroprotectores/uso terapéutico , /efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , /aislamiento & purificación , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cromonas/farmacología , Citocromos c/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Flavonoides/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Morfolinas/farmacología , /clasificación , /citología , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , /aislamiento & purificación , /aislamiento & purificación

RESUMEN

MSTN, IGF-І(insulin-like growth factor-І) and IGF-II (insulin-like growth factor-II) regulate skeletal muscle growth. This study investigated the effects of different dietary intake levels on skeletal muscles. Sheep was randomly assigned to 3 feeding groups: 1) the maintenance diet (M), 2) 1.4 x the maintenance diet (1.4M), and 3) 2.15 x the maintenance diet (2.15M). Before slaughtering the animals, blood samples were collected to measure plasma urea, growth hormone, and insulin concentrations. After slaughtering, the longissimus dorsi, semitendinosus, semimembranosus, gastrocnemius, soleus, and chest muscle were removed to record various parameters, including the mRNA expression levels of MSTN and IGFs, in addition to skeletal muscle fiber diameter and cross-sectional area. The result showed that as dietary intake improved, the mRNA expression levels of MSTN and IGF-II decreased, whereas IGF-Іexpression increased. The mRNA expression levels of MSTN and IGFs were significantly different in the same skeletal muscle under different dietary intake. The skeletal muscle fiber diameter and cross-sectional area increased with greater dietary intake, as observed for the mRNA expression of IGF-І; however, it contrasted to that observed for the mRNA expression of MSTN and IGF-II. In conclusion, dietary intake levels have a certain influence on MSTN and IGFs mRNA expression levels, in addition to skeletal muscle fiber diameter and cross-sectional area. This study contributes valuable information for enhancing the molecular-based breeding of sheep.

Asunto(s)

Dieta/veterinaria , Factor II del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/genética , Carne/análisis , Músculo Esquelético/metabolismo , Miostatina/genética , ARN Mensajero/genética , Alimentación Animal , Animales , Cruzamiento , Quimera/genética , Dieta/métodos , Regulación de la Expresión Génica , Hormona del Crecimiento/sangre , Insulina/sangre , Factor I del Crecimiento Similar a la Insulina/metabolismo , Factor II del Crecimiento Similar a la Insulina/metabolismo , Masculino , Músculo Esquelético/química , Miostatina/metabolismo , ARN Mensajero/metabolismo , Oveja Doméstica , Urea/sangre

RESUMEN

The present study focuses on the neuroprotective effect of glycyrrhizic acid (GA, a major compound separated from Glycyrrhiza Radix, which is a crude Chinese traditional drug) against glutamate-induced cytotoxicity in differentiated PC12 (DPC12) cells. The results showed that GA treatment improved cell viability and ameliorated abnormal glutamate-induced alterations in mitochondria in DPC12 cells. GA reversed glutamate-suppressed B-cell lymphoma 2 levels, inhibited glutamate-enhanced expressions of Bax and cleaved caspase 3, and reduced cytochrome C (Cyto C) release. Exposure to glutamate strongly inhibited phosphorylation of AKT (protein kinase B) and extracellular signal-regulated kinases (ERKs); however, GA pretreatment enhanced activation of ERKs but not AKT. The presence of PD98059 (a mitogen-activated protein/extracellular signal-regulated kinase kinase [MEK] inhibitor) but not LY294002 (a phosphoinositide 3-kinase [PI3K] inhibitor) diminished the potency of GA for improving viability of glutamate-exposed DPC12 cells. These results indicated that ERKs and mitochondria-related pathways are essential for the neuroprotective effect of GA against glutamate-induced toxicity in DPC12 cells. The present study provides experimental evidence supporting GA as a potential therapeutic agent for use in the treatment of neurodegenerative diseases.

Asunto(s)

Antiinflamatorios/uso terapéutico , Ácido Glutámico/toxicidad , Ácido Glicirrínico/uso terapéutico , Fármacos Neuroprotectores/uso terapéutico , Células PC12/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Caspasa 3/aislamiento & purificación , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cromonas/farmacología , Citocromos c/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Flavonoides/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Morfolinas/farmacología , Células PC12/clasificación , Células PC12/citología , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/aislamiento & purificación , Ratas , Proteína X Asociada a bcl-2/aislamiento & purificación

RESUMEN

Mesenchymal stem cells derived from bone marrow (BMSCs) are a population of self-renewing multipotent cells that are capable of differentiating into various cellular lineages, and are widely employed in tissue engineering and cell therapy. Recently, clinical research involving BMSCs has become increasingly popular. In order to conduct appropriate research, it is first necessary to amplify large amounts of functional BMSCs in vitro. However, after several passages of expanding in vitro, the proliferation and differentiation potential of BMSCs gradually decline. To determine whether overexpression of Oct4 or Sox2 might prevent this decline, we transfected Oct4 or Sox2, which are essential for the pluripotency and self-renewal of embryonic stem cells, into BMSCs of Xiaomeishan porcine by a lentivirus. The results showed that overexpression of Sox2 or Oct4 BMSCs in culture media containing a basic fibroblast growth factor resulted in higher proliferation and differentiation compared to controls, suggesting that genetic modification of stemness-related genes is an efficient way to maintain the proliferation and differentiation potential of BMSCs.

Asunto(s)

Adipogénesis , Proliferación Celular , Células Madre Mesenquimatosas/fisiología , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Factores de Transcripción SOXB1/metabolismo , Fosfatasa Alcalina/metabolismo , Animales , Células Cultivadas , Factores de Crecimiento de Fibroblastos/fisiología , Expresión Génica , Células HEK293 , Humanos , Factor 3 de Transcripción de Unión a Octámeros/genética , Osteogénesis , Factores de Transcripción SOXB1/genética , Sus scrofa

RESUMEN

Bovine lactoferrin (bLF) is a member of the transferrin family; it plays an important role in the innate immune response. We identified novel splice variants of the bLF gene in mastitis-infected and healthy cows. Reverse transcription-polymerase chain reaction (RT-PCR) and clone sequencing analysis were used to screen the splice variants of the bLF gene in the mammary gland, spleen and liver tissues. One main transcript corresponding to the bLF reference sequence was found in three tissues in both healthy and mastitis-infected cows. Quantitative real-time PCR analysis showed that the expression levels of the LF gene's main transcript were not significantly different in tissues from healthy versus mastitis-infected cows. However, the new splice variant, LF-AS2, which has the exon-skipping alternative splicing pattern, was only identified in mammary glands infected with Staphylococcus aureus. Sequencing analysis showed that the new splice variant was 251 bp in length, including exon 1, part of exon 2, part of exon 16, and exon 17. We conclude that bLF may play a role in resistance to mastitis through alternative splicing mechanisms.

Asunto(s)

Lactoferrina/genética , Glándulas Mamarias Animales/inmunología , Mastitis Bovina/metabolismo , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus/genética , Empalme Alternativo , Animales , Bovinos , Exones , Femenino , Expresión Génica , Lactoferrina/inmunología , Lactoferrina/metabolismo , Hígado/inmunología , Hígado/metabolismo , Glándulas Mamarias Animales/metabolismo , Glándulas Mamarias Animales/microbiología , Mastitis Bovina/inmunología , Mastitis Bovina/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Bazo/inmunología , Bazo/metabolismo , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/metabolismo , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/inmunología

RESUMEN

The efficacy of endothelin receptor antagonists in protecting against myocardial ischemia/reperfusion (I/R) injury is controversial, and the mechanisms remain unclear. The aim of this study was to investigate the effects of CPU0123, a novel endothelin type A and type B receptor antagonist, on myocardial I/R injury and to explore the mechanisms involved. Male Sprague-Dawley rats weighing 200-250 g were randomized to three groups (6-7 per group): group 1, Sham; group 2, I/R + vehicle. Rats were subjected to in vivo myocardial I/R injury by ligation of the left anterior descending coronary artery and 0.5 percent sodium carboxymethyl cellulose (1 mL/kg) was injected intraperitoneally immediately prior to coronary occlusion. Group 3, I/R + CPU0213. Rats were subjected to identical surgical procedures and CPU0213 (30 mg/kg) was injected intraperitoneally immediately prior to coronary occlusion. Infarct size, cardiac function and biochemical changes were measured. CPU0213 pretreatment reduced infarct size as a percentage of the ischemic area by 44.5 percent (I/R + vehicle: 61.3 ± 3.2 vs I/R + CPU0213: 34.0 ± 5.5 percent, P < 0.05) and improved ejection fraction by 17.2 percent (I/R + vehicle: 58.4 ± 2.8 vs I/R + CPU0213: 68.5 ± 2.2 percent, P < 0.05) compared to vehicle-treated animals. This protection was associated with inhibition of myocardial inflammation and oxidative stress. Moreover, reduction in Akt (protein kinase B) and endothelial nitric oxide synthase (eNOS) phosphorylation induced by myocardial I/R injury was limited by CPU0213 (P < 0.05). These data suggest that CPU0123, a non-selective antagonist, has protective effects against myocardial I/R injury in rats, which may be related to the Akt/eNOS pathway.

Asunto(s)

Animales , Masculino , Ratas , Cardiotónicos/farmacología , Daño por Reperfusión Miocárdica/prevención & control , Pirazoles/farmacología , Receptor de Endotelina A/antagonistas & inhibidores , Receptor de Endotelina B/antagonistas & inhibidores , Análisis de Varianza , Modelos Animales de Enfermedad , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Óxido Nítrico Sintasa de Tipo III/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Distribución Aleatoria , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos

RESUMEN

The efficacy of endothelin receptor antagonists in protecting against myocardial ischemia/reperfusion (I/R) injury is controversial, and the mechanisms remain unclear. The aim of this study was to investigate the effects of CPU0123, a novel endothelin type A and type B receptor antagonist, on myocardial I/R injury and to explore the mechanisms involved. Male Sprague-Dawley rats weighing 200-250 g were randomized to three groups (6-7 per group): group 1, Sham; group 2, I/R + vehicle. Rats were subjected to in vivo myocardial I/R injury by ligation of the left anterior descending coronary artery and 0.5% sodium carboxymethyl cellulose (1 mL/kg) was injected intraperitoneally immediately prior to coronary occlusion. Group 3, I/R + CPU0213. Rats were subjected to identical surgical procedures and CPU0213 (30 mg/kg) was injected intraperitoneally immediately prior to coronary occlusion. Infarct size, cardiac function and biochemical changes were measured. CPU0213 pretreatment reduced infarct size as a percentage of the ischemic area by 44.5% (I/R + vehicle: 61.3 ± 3.2 vs I/R + CPU0213: 34.0 ± 5.5%, P < 0.05) and improved ejection fraction by 17.2% (I/R + vehicle: 58.4 ± 2.8 vs I/R + CPU0213: 68.5 ± 2.2%, P < 0.05) compared to vehicle-treated animals. This protection was associated with inhibition of myocardial inflammation and oxidative stress. Moreover, reduction in Akt (protein kinase B) and endothelial nitric oxide synthase (eNOS) phosphorylation induced by myocardial I/R injury was limited by CPU0213 (P < 0.05). These data suggest that CPU0123, a non-selective antagonist, has protective effects against myocardial I/R injury in rats, which may be related to the Akt/eNOS pathway.

Asunto(s)

Cardiotónicos/farmacología , Antagonistas de los Receptores de la Endotelina A , Antagonistas de los Receptores de la Endotelina B , Daño por Reperfusión Miocárdica/prevención & control , Pirazoles/farmacología , Análisis de Varianza , Animales , Modelos Animales de Enfermedad , Masculino , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Óxido Nítrico Sintasa de Tipo III/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos

RESUMEN

Neuronal apoptosis occurs in the diabetic brain due to insulin deficiency or insulin resistance, both of which reduce the expression of stem cell factor (SCF). We investigated the possible involvement of the activation of the MAPK/ERK and/or AKT pathways in neuroprotection by SCF in diabetes. Male C57/B6 mice (20-25 g) were randomly divided into four groups of 10 animals each. The morphology of the diabetic brain in mice treated or not with insulin or SCF was evaluated by H&E staining and TUNEL. SCF, ERK1/2 and AKT were measured by Western blotting. In diabetic mice treated with insulin or SCF, there was fewer structural change and apoptosis in the cortex compared to untreated mice. The apoptosis rate of the normal group, the diabetic group receiving vehicle, the diabetic group treated with insulin, and the diabetic group treated with SCF was 0.54 ± 0.077 percent, 2.83 ± 0.156 percent, 1.86 ± 0.094 percent, and 1.78 ± 0.095 percent (mean ± SEM), respectively. SCF expression was lower in the diabetic cortex than in the normal cortex; however, insulin increased the expression of SCF in the diabetic cortex. Furthermore, expression of phosphorylated ERK1/2 and AKT was decreased in the diabetic cortex compared to the normal cortex. However, insulin or SCF could activate the phosphorylation of ERK1/2 and AKT in the diabetic cortex. The results suggest that SCF may protect the brain from apoptosis in diabetes and that the mechanism of this protection may, at least in part, involve activation of the ERK1/2 and AKT pathways. These results provide insight into the mechanisms by which SCF and insulin exert their neuroprotective effects in the diabetic brain.

Asunto(s)

Animales , Masculino , Ratones , Apoptosis/efectos de los fármacos , Encéfalo/patología , Diabetes Mellitus Experimental/patología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Neuronas/patología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor de Células Madre/uso terapéutico , Apoptosis/fisiología , Western Blotting , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/enzimología , Hipoglucemiantes/uso terapéutico , Insulina/uso terapéutico , Ratones Endogámicos BALB C , Transducción de Señal , Estreptozocina

RESUMEN

Neuronal apoptosis occurs in the diabetic brain due to insulin deficiency or insulin resistance, both of which reduce the expression of stem cell factor (SCF). We investigated the possible involvement of the activation of the MAPK/ERK and/or AKT pathways in neuroprotection by SCF in diabetes. Male C57/B6 mice (20-25 g) were randomly divided into four groups of 10 animals each. The morphology of the diabetic brain in mice treated or not with insulin or SCF was evaluated by H&E staining and TUNEL. SCF, ERK1/2 and AKT were measured by Western blotting. In diabetic mice treated with insulin or SCF, there was fewer structural change and apoptosis in the cortex compared to untreated mice. The apoptosis rate of the normal group, the diabetic group receiving vehicle, the diabetic group treated with insulin, and the diabetic group treated with SCF was 0.54 +/- 0.077%, 2.83 +/- 0.156%, 1.86 +/- 0.094%, and 1.78 +/- 0.095% (mean +/- SEM), respectively. SCF expression was lower in the diabetic cortex than in the normal cortex; however, insulin increased the expression of SCF in the diabetic cortex. Furthermore, expression of phosphorylated ERK1/2 and AKT was decreased in the diabetic cortex compared to the normal cortex. However, insulin or SCF could activate the phosphorylation of ERK1/2 and AKT in the diabetic cortex. The results suggest that SCF may protect the brain from apoptosis in diabetes and that the mechanism of this protection may, at least in part, involve activation of the ERK1/2 and AKT pathways. These results provide insight into the mechanisms by which SCF and insulin exert their neuroprotective effects in the diabetic brain.

Asunto(s)

Apoptosis/efectos de los fármacos , Encéfalo/patología , Diabetes Mellitus Experimental/patología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Neuronas/patología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor de Células Madre/uso terapéutico , Animales , Apoptosis/fisiología , Western Blotting , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/enzimología , Hipoglucemiantes/uso terapéutico , Insulina/uso terapéutico , Masculino , Ratones , Ratones Endogámicos BALB C , Transducción de Señal , Estreptozocina