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1.
J Assist Reprod Genet ; 37(8): 1807-1814, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32767205

RESUMO

PURPOSE: Reduction in methylenetetrahydrofolate reductase (MTHFR) activity due to genetic variations in the MTHFR gene has been controversially implicated in subfertility in human in vitro fertilization. However, there is no direct gene-knockdown study of embryonic MTHFR to assess its involvement in mammalian preimplantation development. The purpose of this study is to investigate expression profiles and functional roles of MTHFR in bovine preimplantation development. METHODS: Reverse transcription-quantitative PCR (RT-qPCR) and analysis of publicly available RNA-seq data were performed to reveal expression levels of MTHFR during bovine preimplantation development. We knocked down MTHFR by siRNA-mediated RNA interference from the 8- to 16-cell stage and assessed the effects on preimplantation development. RESULTS: The RT-qPCR analysis showed relatively high MTHFR expression at the GV oocyte stage, which was decreased toward the 8- to 16-cell stage and then slightly restored at the blastocyst stage. Public data-based analysis also showed the similar pattern of expression with substantial embryonic expression at the blastocyst stage. MTHFR knockdown reduced the blastocyst rate (P < 0.01) and the numbers of total (P < 0.0001), trophectoderm (P < 0.0001), and inner cell mass (P < 0.001) cells. CONCLUSION: The results indicate that embryonic MTHFR is indispensable for normal blastocyst development. The findings provide insight into the debatable roles of MTHFR in fertility and may be applicable for the improvement of care for early embryos via modulation of surrounding folate-related nutritional conditions in vitro and/or in utero, depending on the parental and embryonic MTHFR genotype.


Assuntos
Blastocisto/enzimologia , Desenvolvimento Embrionário/genética , Fertilidade/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Animais , Blastocisto/metabolismo , Blastocisto/ultraestrutura , Bovinos , Feminino , Fertilidade/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Oócitos/enzimologia , Oócitos/crescimento & desenvolvimento , Oócitos/ultraestrutura , RNA Interferente Pequeno
2.
Artigo em Inglês | MEDLINE | ID: mdl-29757463

RESUMO

Depression is one of the most common psychiatric diseases and is commonly comorbid with type 1 or 2 diabetes mellitus (DM). However, the pathophysiology underlying the depressive state in DM remains poorly understood. Animal models are useful tools to investigate the association between depression and DM. In the present study we investigated whether the Spontaneously Diabetic Torii (SDT) fatty rat, a novel animal model of type 2 DM, shows depression-related features. We assessed depression-like behaviour, hyperactivation of the hypothalamic-pituitary-adrenal (HPA) axis, and neurotransmitter levels in the brain. Behaviour was evaluated using a forced swimming test, and the HPA axis was evaluated with changes in plasma corticosterone levels after a swimming stress exposure or dexamethasone challenge. In addition, serotonin (5-hydroxytryptamine; 5-HT), noradrenaline, glutamate and γ-aminobutyric acid (GABA) concentrations in the frontal cortex, hippocampus and brain stem were measured. In the forced swimming test, SDT fatty rats exhibited increased duration of immobility compared with control Sprague-Dawley (SD) rats. Moreover, basal corticosterone levels were significantly elevated in SDT fatty compared with control SD rats. However, there were no stress-induced increases or changes in dexamethasone-induced suppression of corticosterone in SDT fatty compared with control SD rats. Furthermore, there were significant changes in 5-HT concentrations in the prefrontal cortex, and in GABA and glutamate concentrations in the hippocampus in SDT fatty compared with controls. The results of the present study suggest that the SDT fatty rat may be an appropriate model for diabetes with comorbid depression associated with neurotransmitter impairments and aberrant basal HPA hyperactivity.

3.
J Reprod Dev ; 64(2): 199-202, 2018 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-29353869

RESUMO

Bovine preimplantation embryos exhibit dramatic biological changes between before and after the 8-16-cell stage. Here we report a simple lipofection method to transfect siRNA into bovine 8-16-cell stage embryos using zona removal and the well-of-the-well (WOW) culture system. Bovine one-cell embryos produced in vitro were freed from the zona pellucida and cultured up to the 8-16-cell stage in WOW dishes. The 8-16-cell embryos were lipofected with siRNA and the transfection efficiency was assessed at 48 h of transfection. Lipofection with a red fluorescent non-targeting siRNA revealed the importance of zona removal for transfection of siRNA into embryos. Using this method, we knocked down the methionine adenosyltransferase 2A (MAT2A) gene, achieving a significant reduction in MAT2A expression (P < 0.05) concomitant with the marked inhibition of blastocyst development. Our proposed method, tentatively named 'Octo-lipofection', may be useful to analyze gene functions in bovine preimplantation embryos without expensive equipment and skill-intensive techniques.


Assuntos
Ectogênese , Mórula/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transfecção/métodos , Animais , Bovinos , Ectogênese/efeitos dos fármacos , Técnicas de Cultura Embrionária , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Indicadores e Reagentes/farmacologia , Lipídeos/farmacologia , Masculino , Metionina Adenosiltransferase/antagonistas & inibidores , Metionina Adenosiltransferase/genética , Metionina Adenosiltransferase/metabolismo , Mórula/efeitos dos fármacos , Transfecção/veterinária , Zona Pelúcida/fisiologia
4.
J Pharmacol Sci ; 130(2): 136-8, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26883454

RESUMO

Some nutrients, such as carbohydrate, fat and protein, are known to stimulate satiety. However, the effect of sn-2-monoacylglycerol (2-MG), one of the digestive products of triglycerides, on food intake is still unclear. In the present study, the effects of 2-MG on food intake and diarrhea were evaluated and compared with long-chain fatty acid (LCFA) in rats by intrajejunal infusion. Intrajejunal infusion of 2-MG reduced food intake. In addition, 2-MG did not induce diarrhea at the condition that it comparably reduced food intake as compared with LCFA. These results suggest that 2-MG stimulates satiety without inducing diarrhea, different from LCFA.


Assuntos
Diarreia/etiologia , Ingestão de Alimentos/efeitos dos fármacos , Monoglicerídeos/farmacologia , Saciação/efeitos dos fármacos , Animais , Depressão Química , Ácidos Graxos/administração & dosagem , Ácidos Graxos/farmacologia , Ácidos Graxos/fisiologia , Jejuno , Masculino , Monoglicerídeos/administração & dosagem , Monoglicerídeos/fisiologia , Ratos Sprague-Dawley , Estimulação Química
5.
J Pharmacol Sci ; 128(3): 150-7, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26215699

RESUMO

AIM: Monoacyglycerol acyltransferases (MGATs) are known to play important roles in intestinal TG absorption. In contrast, the role of MGATs in the liver is still unclear. We investigated the effects of JTP-103237, a novel MGAT inhibitor, on hepatic MGAT activity and hepatic lipid metabolism. RESULTS: JTP-103237 reduced hepatic triglyceride content and hepatic MGAT activity in a high sucrose very low fat (HSVLF) diet induced fatty liver model. Interestingly, JTP-103237 suppressed not only triglyceride (TG) and diacylglycerol (DG) synthesis, but also fatty acid (FA) synthesis (de novo lipogenesis) in this model. JTP-103237 also suppressed lipogenesis-related gene expression, such as sterol regulatory element-binding protein 1-c. Moreover, JTP-103237 decreased plasma glucose levels and total cholesterol and reduced the accumulation of epididymal fats in HSVLF diet fed mice. CONCLUSION: In the present study, JTP-103237 prevented carbohydrate-induced fatty liver and suppressed both TG synthesis and de novo lipogenesis, suggesting MGAT inhibitor may prevent carbohydrate-induced metabolic disorders, including NAFLD, obesity and diabetes.


Assuntos
Aciltransferases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Ácidos Graxos/biossíntese , Fígado Gorduroso/metabolismo , Lipogênese/efeitos dos fármacos , Fígado/metabolismo , Piperazinas/farmacologia , Triazóis/farmacologia , Triglicerídeos/biossíntese , Aciltransferases/metabolismo , Aciltransferases/fisiologia , Animais , Antígenos de Bactérias , Proteínas de Bactérias , Diglicerídeos/biossíntese , Modelos Animais de Doenças , Fígado Gorduroso/prevenção & controle , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/genética , Absorção Intestinal/efeitos dos fármacos , Lipogênese/genética , Masculino , Camundongos Endogâmicos C57BL , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Triglicerídeos/metabolismo
6.
Br J Nutr ; 111(2): 247-53, 2014 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-23830034

RESUMO

An adequate immune system is required to prevent diarrhoea in neonates, and IgA provides protection against microbial antigens on mucosal surfaces. Although ß-carotene supplementation has been expected to enhance the retinoic acid (RA)-mediated immune response in neonates, the exact mechanism of the enhancement of mucosal IgA production in the small intestine by ß-carotene is still unclear. In the present study, we investigated the effect of supplemental ß-carotene on the concentrations of IgA, the numbers of IgA antibody-secreting cells (ASC) and the mRNA expressions of IgA C-region, CCL25, retinoid X receptor (RXR) α, retinoic acid receptor (RAR) α and RARγ in the jejunum and ileum of weanling mice. Weanling mice were fed rodent feed or 50 mg/kg ß-carotene-supplemented rodent feed for 7, 14 or 21 d. The concentrations of IgA and the numbers of IgA ASC in the jejunum and ileum of mice increased markedly with age, and supplemental ß-carotene increased the concentrations of IgA, the numbers of IgA ASC and the mRNA expressions of IgA C-region, CCL25 and RARγ in the jejunum after 14 and 21 d of treatment. Supplemental ß-carotene increased the numbers of IgA ASC in the ileum after 14 and 21 d of treatment, but the concentrations of IgA in the ileum were not affected by ß-carotene supplementation. The mRNA expressions of RXRα and RARα in the jejunum and those of RXRα and RARγ in the ileum after 21 d of treatment were enhanced by ß-carotene supplementation. These results indicate that ß-carotene supplementation in weanling mice is effective to enhance mucosal IgA induction in the jejunum or ileum and that the effects are mainly due to the RA-mediated immune response.


Assuntos
Suplementos Nutricionais , Íleo/metabolismo , Imunoglobulina A/metabolismo , Jejuno/metabolismo , beta Caroteno/farmacologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Regulação da Expressão Gênica/efeitos dos fármacos , Íleo/efeitos dos fármacos , Imunoglobulina A/sangue , Jejuno/efeitos dos fármacos , Camundongos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Desmame
7.
J Reprod Dev ; 60(4): 317-23, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24899099

RESUMO

Intracellular calcium homeostasis is essential for proper cell function. We investigated the effects of heat shock on the development of and the intracellular Ca2+ levels in bovine preimplantation embryos in vitro and the effects of calcitonin (CT), a receptor-mediated Ca2+ regulator, on heat shock-induced events. Heat shock (40.5 C for 10 h between 20 and 30 h postinsemination) of in vitro-produced bovine embryos did not affect the cleavage rate; however, it significantly decreased the rates of development to the 5- to 8-cell and blastocyst stages as compared with those of the control cultured for the entire period at 38.5 C (P < 0.05). The relative intracellular Ca2+ levels at the 1-cell stage (5 h after the start of heat shock), as assessed by Fluo-8 AM, a fluorescent probe for Ca2+, indicated that heat shock significantly lowered the Ca2+ level as compared with the control level. Semiquantitative reverse transcription PCR and western blot analyses revealed the expression of CT receptor in bovine preimplantation embryos. The addition of CT (10 nM) to the culture medium ameliorated the heat shock-induced impairment of embryonic development beyond the 5- to 8-cell stage. The Ca2+ level in the heat-shocked embryos cultured with CT was similar to that of the control embryos, suggesting that heat shock lowers the Ca2+ level in fertilized embryos in vitro and that a lower Ca2+ level is implicated in heat shock-induced impairment of embryonic development. Intracellular Ca2+ -mobilizing agents, e.g., CT, may effectively circumvent the detrimental effects of heat shock on early embryonic development.


Assuntos
Cálcio/metabolismo , Embrião de Mamíferos/metabolismo , Temperatura Alta , Receptores da Calcitonina/metabolismo , Estresse Fisiológico , Animais , Calcitonina , Bovinos , Camundongos , Oócitos/metabolismo
8.
J Reprod Dev ; 58(1): 38-43, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22450283

RESUMO

One-carbon metabolism (OCM) can be seen as integrated metabolic pathways centered on the metabolism of two nutritional substances, folate and methionine. Mammalian oocytes and preimplantation embryos express almost all enzymes that participate in OCM, suggesting that they can independently metabolize OCM nutrients. A deficiency or excess of OCM nutrients and their metabolites during in vitro culture affects preimplantation development of mammalian embryos. Recent in vivo studies have demonstrated that specific OCM dietary interventions during the periconceptional (mainly oocyte growth and preimplantation) period can cause epigenetic alterations in DNA of offspring and program the long-term consequences in their health in adulthood. The epigenetic processes are likely to be implicated in the effects of OCM nutrients; however, understanding their effects at the level of specific genes and their implications in assisted reproductive technology will require further investigations.


Assuntos
Blastocisto/metabolismo , Ácido Fólico/metabolismo , Metionina/metabolismo , Animais , Bovinos , Desenvolvimento Embrionário , Epigênese Genética , Feminino , Humanos , Masculino , Camundongos , Oócitos/metabolismo , Ratos
9.
J Reprod Dev ; 58(1): 91-7, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22052008

RESUMO

The roles of methionine metabolism in bovine preimplantation embryo development were investigated by using ethionine, an antimetabolite of methionine. In vitro produced bovine embryos that had developed to the 5-cell stage or more at 72 h after the commencement of in vitro fertilization (IVF) were then cultured until day 8 (IVF = day 0) in medium supplemented with 0 (control), 1, 5 and 10 mM ethionine. Compared with the blastocyst development in the control (40.0%), ethionine at 10 mM almost completely blocked blastocyst development (1.1%, P<0.001), and this concentration was used in the following experiments. Methionine added at the same concentration (10 mM, a concentration control of ethionine) did not cause such an intense developmental inhibition. Development to the compacted morula stage on day 6 was not affected by 10 mM ethionine treatment. S-adenosylmethionine (SAM) added to the ethionine treatment partly restored the blastocyst development. Semiquantitative reverse transcription-polymerase chain reaction analysis of cell lineage-related transcription factors in day 6 compacted morulae showed that the expressions of NANOG and TEAD4 were increased by ethionine treatment relative to the control (P<0.01). Furthermore, immunofluorescence analysis of 5-methylcytosine revealed that DNA was hypomethylated in the ethionine-treated day 6 morulae compared with the control (P<0.001). These results demonstrate that the disruption of methionine metabolism causes impairment of the morula-to-blastocyst transition during bovine preimplantation development in part via SAM deficiency, indicating the indispensable roles of methionine during this period. The disruption of methionine metabolism may cause hypomethylation of DNA and consequently lead to the altered expression of developmentally important genes, which then results in the impairment of blastocyst development.


Assuntos
Blastocisto/metabolismo , Desenvolvimento Embrionário/fisiologia , Metionina/metabolismo , Mórula/metabolismo , Animais , Antimetabólitos/farmacologia , Bovinos , Metilação de DNA/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Etionina/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Gravidez , Fatores de Transcrição/metabolismo
10.
J Vet Med Sci ; 84(3): 330-337, 2022 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-35082197

RESUMO

Diabetes mellitus (DM) and obesity are associated with neurodegenerative diseases such as Alzheimer's disease and psychiatric disorders such as major depression. In this study, we investigated pathophysiological changes in the brains of female Spontaneously Diabetic Torii (SDT) fatty rats with diabetes and obesity. Brains of Sprague-Dawley (SD), SDT and SDT fatty rats were collected at 58 weeks of age. The parietal cortical thickness was measured and the number of pyramidal cells in the hippocampal cornu ammonis 1 and 3 (CA1 and CA3) and the number of granule cells in the dentate gyrus (DG) regions were counted. The area of glial fibrillary acidic protein (GFAP) positivity in CA1, CA3 and DG regions were measured. The parietal cortical thickness and the number of cells in CA3 and DG regions of SDT and SDT fatty rats did not show obvious changes. On the other hand, in the CA1 region, the number of cells in SDT rats and SDT fatty rats was significantly lower than that in SD rats, and that in SDT fatty rats was significantly lower than that in SDT rats. The GFAP-positive area in SDT fatty rats was significantly reduced compared to that in SD rats only in the DG region. Preliminarily result showed that the expression of S100a9, an inflammation-related gene, was increased in the brains of SDT fatty rats. These results suggest that female SDT fatty rat may exhibit central nervous system diseases due to obesity and DM.


Assuntos
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Doenças dos Roedores , Animais , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Tipo 2/veterinária , Modelos Animais de Doenças , Feminino , Obesidade/complicações , Obesidade/veterinária , Ratos , Ratos Sprague-Dawley
11.
Br J Nutr ; 105(1): 24-30, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20727240

RESUMO

Mortality of neonates continues to be a major problem in humans and animals. IgA provides protection against microbial antigens at mucosal surfaces. Although ß-carotene supplementation has been expected to enhance retinoic acid-mediated immune response in neonates, the exact mechanism by which ß-carotene enhances IgA production is still unclear. We investigated the effect of supplemental ß-carotene for maternal mice during pregnancy and lactation on IgA antibody-secreting cells (ASC) in mammary gland and guts and on IgA transfer from milk to neonatal mice. Pregnant mice were fed untreated or 50 mg/kg ß-carotene-supplemented diets from 6·5 d postcoitus (dpc) to 14 d postpartum (dpp). Supplemental ß-carotene increased the numbers of IgA ASC in mammary gland (P < 0·05) and ileum (P < 0·001), and also mRNA expression of IgA C-region in ileum (P < 0·05) of maternal mice at 14 dpp, but few IgA ASC were detected in mammary gland at 17·5 dpc. IgA concentration in stomach contents, which represents milk IgA level, was significantly higher (P < 0·01) in neonatal mice born to ß-carotene-supplemented mothers at 7 and 14 dpp, and IgA concentration in serum, stomach contents and faeces increased (P < 0·001) drastically with age. These results suggest that ß-carotene supplementation for maternal mice during pregnancy and lactation is useful for enhancing IgA transfer from maternal milk to neonates owing to the increase in IgA ASC in mammary gland and ileum during lactation.


Assuntos
Células Produtoras de Anticorpos/efeitos dos fármacos , Imunoglobulina A/metabolismo , Lactação/imunologia , Glândulas Mamárias Animais/imunologia , Fenômenos Fisiológicos da Nutrição Materna/imunologia , Leite/imunologia , beta Caroteno/farmacologia , Animais , Animais Recém-Nascidos , Animais Lactentes , Células Produtoras de Anticorpos/citologia , Células Produtoras de Anticorpos/imunologia , Suplementos Nutricionais , Fezes/química , Feminino , Conteúdo Gastrointestinal/química , Íleo/imunologia , Imunoglobulina A/genética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Gravidez , RNA Mensageiro/metabolismo , beta Caroteno/sangue , beta Caroteno/metabolismo
12.
Phytother Res ; 25(5): 654-8, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21031635

RESUMO

The present study was conducted to clarify the effects of coumestrol administration on Ca metabolism during pregnancy and in lactating mice. From 6.5 to 16.5 days post coitus (dpc), pregnant mice were administered coumestrol at 200 µg/kg body weight/day. The duodenum, jejunum and blood samples were obtained at 17.5 dpc or 10 days after parturition (dap). Coumestrol administration decreased alkaline phosphatase (ALP) activity and mRNA expression of IAP and estrogen responsive genes, c-fos and vascular endothelial growth factor (VEGF), in the duodenum and jejunum of pre-delivery mice. In lactating mice, the ALP activity and mRNA expression of IAP were not changed, although coumestrol administration decreased mRNA expression of c-fos in the duodeum and VEGF in the jejunum. Coumestrol did not affect serum Ca and the expression of vitamin D receptor protein in the duodenum and jejunum. Thus, coumestrol administration during pregnancy may decrease the mRNA expression of IAP and the ALP activity in the intestine of the pre-delivery mice through ERα, but coumestrol had little effect on intestinal ALP activity at 10 days after parturition.


Assuntos
Fosfatase Alcalina/metabolismo , Cálcio/sangue , Cumestrol/farmacologia , Intestinos/enzimologia , Fitoestrógenos/farmacologia , Fosfatase Alcalina/efeitos dos fármacos , Fosfatase Alcalina/genética , Animais , Cálcio/metabolismo , Cumestrol/administração & dosagem , Duodeno/efeitos dos fármacos , Duodeno/enzimologia , Feminino , Humanos , Imuno-Histoquímica , Intestinos/efeitos dos fármacos , Jejuno/efeitos dos fármacos , Jejuno/enzimologia , Lactação/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Fitoestrógenos/administração & dosagem , Gravidez , Proteínas Proto-Oncogênicas c-fos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Calcitriol/genética , Receptores de Calcitriol/metabolismo , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/genética
13.
J Vet Med Sci ; 80(9): 1385-1391, 2018 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-30012919

RESUMO

In recent years, a relationship between diabetes and neurodegenerative diseases, such as Parkinson's disease, Alzheimer disease or depression, has been proposed. In this study, pathophysiological changes in the brain, especially in the hippocampus, of male SDT fatty rats with obesity and hyperglycemia were investigated. Brains of SD rats and SDT fatty rats were collected at 32 and 58 weeks of age, and parietal cortical thickness and number of pyramidal cells in the hippocampal cornu ammonis 1 and 3 (CA1 and CA3) regions were measured. At 58 weeks of age, the parietal cortical thickness and number of pyramidal cells in the hippocampal CA1 and CA3 regions were lower in SDT fatty rats than in age-matched SD rats. Measurements of mRNA in rat brains at 58 weeks of age showed that the expression of genes related to inflammatory responses (S100a9, TNFα, NF-κB) was elevated in SDT fatty rats. From the aforementioned results, changes suggestive of brain atrophy and impairment in cognitive function were observed in male SDT fatty rat brains.


Assuntos
Encéfalo/patologia , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/fisiopatologia , Modelos Animais de Doenças , Animais , Encéfalo/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley
14.
Int J Endocrinol ; 2018: 9065690, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30344606

RESUMO

In patients with diabetes mellitus (DM), impairments of circadian rhythms, including the sleep-wake cycle, blood pressure, and plasma melatonin concentrations, are frequently observed. Animal models of DM are also reported to show aberrant circadian rhythms. However, the changes in the circadian rhythms of plasma soluble substances, including melatonin, in diabetic animals are controversial. In the present study, we investigated the circadian rhythms of spontaneous locomotor activity, metabolic parameters (plasma glucose, triglyceride, and total cholesterol), and plasma melatonin concentrations in Spontaneously Diabetic Torii (SDT) fatty rats, a novel animal model of type 2 DM. Although SDT fatty rats exhibited low locomotor activity in the dark phase, no phase shifts were observed. The circadian variations of plasma metabolic parameters were more apparent in the SDT fatty rats compared with control Sprague-Dawley (SD) rats. The circadian rhythms of plasma melatonin concentrations were significantly impaired in SDT fatty rats. To get an insight into the mechanism underlying the impaired melatonin secretion in SDT fatty rats, the expression of arylalkylamine N-acetyltransferase (Aanat) and acetylserotonin O-methyltransferase (Asmt) mRNA, which encode the rate-limiting enzymes for melatonin synthesis, was investigated in the pineal gland. There were no significant differences in Aanat and Asmt expression between the control SD and SDT fatty rats. These results suggest that SDT fatty rats show impaired circadian rhythms and dysregulated melatonin secretion.

15.
J Vet Med Sci ; 80(6): 878-885, 2018 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-29643297

RESUMO

Nonalcoholic steatohepatitis (NASH) is a progressive liver disease, and some patients develop hepatic cirrhosis/carcinoma. Animal models play key roles in the development of new therapies for NASH. In this study, the pharmacological effects of metformin and pioglitazone were investigated in female Spontaneously Diabetic Torii (SDT) fatty rats to verify the utility of this model. The anti-diabetic drugs were administered to SDT fatty rats fed a cholesterol-enriched diet from 4 to 25 weeks, and changes in food intake, body weight, and blood chemistry parameters were evaluated every 4 weeks. The hepatic lipid content, mRNA expression in relation to lipid synthesis, inflammation, and fibrosis, and histopathological analyses were performed at 25 weeks. Pioglitazone improved hyperglycemia, hyperlipidemia, and abnormalities in hepatic parameters. The insulin levels were lower than those in the control rats before 16 weeks. Plasma glucose levels in the metformin-treated rats were lower than those in the control rats, and plasma alanine aminotransferase levels temporarily decreased. The lipid content and some mRNA expression in relation to fibrosis in the liver decreased with pioglitazone treatment, and the mRNA expression of microsomal triglyceride transfer protein increased. Hepatic fibrosis observed in the SDT fatty rats improved with pioglitazone treatment; however, the effect with metformin treatment was partial. These results in both drugs are in line with results in the human study, suggesting that the SDT fatty rat is useful for developing new anti-NASH drugs that show potential to regulate glucose/lipid metabolism.


Assuntos
Modelos Animais de Doenças , Hipoglicemiantes/uso terapêutico , Metformina/uso terapêutico , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Tiazolidinedionas/uso terapêutico , Animais , Colesterol , Dieta , Ingestão de Alimentos , Feminino , Tamanho do Órgão , Pioglitazona , RNA Mensageiro/metabolismo , Ratos
16.
Anim Sci J ; 89(8): 1102-1106, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29808628

RESUMO

Data from 26 Japanese Black cows were collected to clarify the effects of supplemental ß-carotene on colostral immunoglobulin (Ig) and plasma ß-carotene and Ig in the cows. Cows were assigned to control or ß-carotene groups from 21 days before the expected calving date to 60 days after parturition. Supplemental ß-carotene was provided at 500 mg/day in the ß-carotene group. Supplemental ß-carotene drastically increased plasma ß-carotene concentrations in the cows from parturition to 60 days after parturition, and plasma ß-carotene concentrations in the control and ß-carotene groups at parturition were 202 and 452 µg/dl, respectively. Supplemental ß-carotene had no effects on plasma IgG1 , IgA or IgM concentrations at parturition. Supplemental ß-carotene increased colostral IgG1 concentrations in the cows, but colostral ß-carotene, IgA and IgM concentrations were not affected by supplemental ß-carotene. These results indicate that supplemental ß-carotene is effective to enhance colostral IgG1 concentrations and plasma ß-carotene concentrations in Japanese Black cows.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal/imunologia , Colostro/imunologia , Colostro/metabolismo , Suplementos Nutricionais , Imunoglobulina G/metabolismo , beta Caroteno/administração & dosagem , beta Caroteno/sangue , Animais , Bovinos , Dieta , Feminino , Imunoglobulinas/sangue , Imunoglobulinas/metabolismo , Parto/sangue , Parto/imunologia , Gravidez , beta Caroteno/farmacologia
17.
Sci Rep ; 7(1): 3800, 2017 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-28630431

RESUMO

Methionine adenosyltransferase (MAT) is involved in folate-mediated one-carbon metabolism, which is essential for preimplantation embryos in terms of both short-term periconceptional development and long-term phenotypic programming beyond the periconceptional period. Here, our immunofluorescence analysis of bovine oocytes and preimplantation embryos revealed the consistent expression of MAT2A (the catalytic subunit of the ubiquitously expressed-type of MAT isozyme) during this period. Addition of the MAT2A inhibitor FIDAS to the culture media of bovine preimplantation embryos reduced their blastocyst development, revealing the particular importance of MAT2A in successful blastocyst development. Exploration of MAT2A-associated genomic regions in bovine blastocysts using chromatin immunoprecipitation and sequencing (ChIP-seq) identified candidate MAT2A-associated genes implicated not only in short-term periconceptional embryo development, but also in long-term phenotypic programming during this period in terms of growth, metabolism, and immune functions. These results suggest the critical involvement of MAT2A in the periconceptional period in life-long programming of health and disease as well as successful preimplantation development.


Assuntos
Blastocisto/enzimologia , Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Metionina Adenosiltransferase , Animais , Blastocisto/citologia , Bovinos , Desenvolvimento Embrionário/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Metionina Adenosiltransferase/antagonistas & inibidores , Metionina Adenosiltransferase/biossíntese , Metionina Adenosiltransferase/genética
18.
J Pharmacol Toxicol Methods ; 88(Pt 2): 160-166, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28970182

RESUMO

INTRODUCTION: To establish an animal model for diabetic peripheral neuropathy (DPN) at an earlier stage, we performed functional and pathophysiological evaluations in Spontaneously Diabetic Torii (SDT) fatty rats before 16weeks of age. METHODS: Male SDT fatty rats were treated with vehicle or phlorizin (100 to 150mg/kg/day) from 5 to 16weeks. Sprague-Dawley (SD) rats were used as age-matched controls. Body weights and biochemical parameters were measured over time. During the treatment period, the sensory and motor nerve conduction velocity (SNCV and MNCV) of the sciatic nerve, blood pressure, pupil size, and electrocardiograms were measured. At 16weeks, the rats were sacrificed and sural nerves and intraepidermal nerves were sampled for histological studies, electron microscopic analysis and assessments of nerve fiber density. RESULTS: Functional abnormalities, such as delays of SNCV, increase of blood pressure, reduced pupillary reactivity, and decrease of the coefficient of variance of R-R intervals were observed in SDT fatty rats. Histopathologically, decreased intraepidermal nerve fiber density, mitochondrial abnormalities of small myelinated fibers, and vacuolation and mitochondrial swelling of unmyelinated fibers were found in SDT fatty rats. These changes were prevented by well-controlled blood glucose with phlorizin treatment. DISCUSSION: Male SDT fatty rats can help future work on DPN in diabetes with obesity, since this rat exhibited functional and pathological abnormalities in somatic and autonomic nerve from an early stage of diabetes.


Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Neuropatias Diabéticas/fisiopatologia , Modelos Animais de Doenças , Obesidade/fisiopatologia , Animais , Diabetes Mellitus Experimental/genética , Neuropatias Diabéticas/genética , Masculino , Obesidade/genética , Ratos , Ratos Sprague-Dawley , Ratos Transgênicos
19.
Anim Sci J ; 88(4): 653-658, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27592519

RESUMO

Data from 18 ß-carotene-deficient Japanese Black cows were collected to clarify the effects of feeding ß-carotene-enriched dry carrots on ß-carotene status and colostral immunoglobulin (Ig) in cows. Cows were assigned to control or carrot groups from 3 weeks before the expected calving date to parturition, and supplemental ß-carotene from dry carrots was 138 mg/day in the carrot group. Plasma ß-carotene concentrations in the control and carrot groups at parturition were 95 and 120 µg/dL, and feeding dry carrots slightly improved plasma ß-carotene at parturition. Feeding dry carrots increased colostral IgA concentrations in cows and tended to increase colostral IgG1 , but colostral IgM, IgG2 , ß-carotene and vitamin A were not affected by the treatment. Feeding dry carrots had no effects on plasma IgG1 , IgA and IgM concentrations in cows, but plasma IgG1 concentrations decreased rapidly from 3 weeks before the expected calving date to parturition. These results indicate that feeding ß-carotene-enriched dry carrots is effective to enhance colostral IgA and IgG1 concentrations in ß-carotene-deficient cows.


Assuntos
Ração Animal , Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Colostro/imunologia , Daucus carota , Alimentos Fortificados , Imunoglobulina A Secretora/metabolismo , Imunoglobulina G/metabolismo , beta Caroteno/administração & dosagem , Animais , Bovinos , Colostro/metabolismo , Feminino , Gravidez , beta Caroteno/sangue , beta Caroteno/deficiência
20.
Anim Sci J ; 87(4): 536-40, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26259528

RESUMO

This study was conducted to clarify the relationships among immunoglobulin (Ig)M, IgG, IgA, ß-carotene, vitamin A and α-tocopherol contents in colostrum of 24 Japanese Black multiparous cows in order to evaluate the role of IgM on colostral IgG and IgA production. Compared with colostral IgG, colostral IgM and IgA were very low but varied widely. There was positive correlation between colostral IgM and IgG, but colostral IgM was not related with colostral IgA. There was no relationship between colostral IgM and age of cows, although colostral IgG was increased with aging. There were positive correlations among colostral ß-carotene, vitamin A and α-tocopherol and these vitamins were positively correlated with colostral IgM and IgG. These results indicate that fat-soluble vitamins may affect colostral IgG and IgM in cows and colostral IgG increases with the increase of colostral IgM.


Assuntos
Colostro/química , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Imunoglobulina M/metabolismo , Envelhecimento/metabolismo , Animais , Bovinos , Feminino , Vitamina A/metabolismo , alfa-Tocoferol/metabolismo , beta Caroteno/metabolismo
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