RESUMO
To achieve appropriate blood pressure control in the treatment of hypertension in Japan, this study examined the relationship between office blood pressure and actual antihypertensive drug use in general hospitals following the promulgation of the guidelines for hypertension (JSH2019). This study focused on blood pressure levels and drug use in outpatients on antihypertensive treatment from June to July 2020. The subjects were 2,537 patients classified into four groups based on their medical history, patients with: hypertension only; hypertension and cardiovascular disease; hypertension and dyslipidaemia; and hypertension and diabetes mellitus. The results showed a significant difference in systolic blood pressure (SBP) between patients with hypertension only and those with hypertension and cardiovascular disease (138.3±17.9 mmHg vs 135.6±19.9 mmHg, p<0.05). Regarding actual drug use, it was found that diuretics were prescribed more frequently in patients with hypertension and cardiovascular disease than in those with hypertension alone (15.5% vs 37.9%, p<0.05), even though the number of drugs for hypertension did not differ significantly. In addition, the dose of diuretics was greater only in patients with cardiovascular disease. These results show the actual drug use and blood pressure for each comorbidity. Furthermore, they suggest that the results of antihypertensive treatment may differ by changing the combination and dosage of antihypertensive drugs without changing the number of antihypertensive drugs used. The study also shows the problem of using less diuretics depending on the risk the patient has, and solving the problem may lead to achieving further antihypertensive goals.
Assuntos
Doenças Cardiovasculares , Hipertensão , Humanos , Anti-Hipertensivos/uso terapêutico , Anti-Hipertensivos/farmacologia , Pressão Sanguínea , Doenças Cardiovasculares/induzido quimicamente , Hipertensão/tratamento farmacológico , Diuréticos/uso terapêuticoRESUMO
Antimicrobials should be used appropriately to minimise the risk of resistant strains arising in association with overuse. De-escalation of antimicrobial therapy is one strategy used to ensure appropriate use, but its safety and efficacy in burn patients are unclear. The aim of this study was to evaluate the safety and efficacy of de-escalation therapy for treating infections in burn patients. This retrospective cohort study investigated patients admitted to our intensive care unit with burns and treated for infection between October 1, 2013, and September 30, 2020. Patients were classified into a de-escalation group (Group D) comprising patients treated with empiric antimicrobial therapy followed by de-escalation and a non-de-escalation group (Group ND) comprising patients who did not undergo de-escalation. Characteristics and outcomes were compared between groups. Forty-three patients met the inclusion criteria, including 15 patients in Group D and 28 patients in Group ND. Bacterial species commonly detected in these patients were Corynebacterium spp. (17.3%), Pseudomonas aeruginosa (16.1%), and Staphylococcus aureus (9.6%) . No inter-group difference was seen in 28-day mortality (6.7% vs 21.4%, p =0.391). Multidrug-resistant strains were detected significantly less frequently in Group D (13.0%) than in Group ND (26.1%, p =0.003). De-escalation was associated with use of two or more antimicrobials as empiric antimicrobial therapy. As the use of de-escalation in infection treatment did not impact 28-day mortality, de-escalation might be safe for treating infections in burn patients.
Assuntos
Anti-Infecciosos , Queimaduras , Humanos , Estudos Retrospectivos , Antibacterianos/uso terapêutico , Anti-Infecciosos/uso terapêutico , Unidades de Terapia Intensiva , Queimaduras/tratamento farmacológicoRESUMO
In the present work, taste masked particles of acetaminophen (AAP), a highly soluble bitter tasting drug, were developed and ODT containing the taste masked particles were prepared. Taste masked particles of AAP were prepared using different amounts of tetraglycerol polyricinoleate (TGPR) and Eudragit ®E100. Although the drug content ratio and drug recovery decreased with increasing TGPR, drug release from AAP-CR100 particles containing a large amount of TGPR was mostly suppressed for 2 min. Hence, AAP-CR100 was incorporated into ODT as taste masked particles for AAP. Three major disintegrants were used for ODT, and it was confirmed that the tensile strength of all formulations showed applicable hardness for handling. The AAP-CR100-CP(40) formulation containing crospovidone showed the shortest disintegration time and the drug release from AAP-CR100-CP(40) into pH 6.8 test solution was suppressed compared with commercial AAP tablets. Because the drug release from AAP-CR100-CP(40) into the pH 1.2 test solution was rapid, it was suggested that drug release from AAP-CR100-CP(40) is suppressed in the oral cavity, and the drug is released promptly in the stomach. Thus AAP-CR100-CP(40) may be useful as an ODT in which the dissolution of AAP in the oral cavity is suppressed.
Assuntos
Acetaminofen/administração & dosagem , Excipientes/química , Povidona/química , Paladar , Acetaminofen/química , Acrilatos/química , Administração Oral , Liberação Controlada de Fármacos , Dureza , Concentração de Íons de Hidrogênio , Polímeros/química , Comprimidos , Resistência à TraçãoRESUMO
WHAT IS KNOWN AND OBJECTIVE: Anticholinergic drugs are associated with risks of falls, confusion and cognitive dysfunction. However, the effect of anticholinergic drug use on rehabilitation outcomes after a stroke is poorly documented. We therefore aimed to establish whether the anticholinergic load was associated with functional recovery among geriatric patients convalescing after stroke. METHOD: Consecutive geriatric stroke patients admitted and discharged from a convalescence rehabilitation ward between 2010 and 2016 were included in this retrospective cohort study. Anticholinergic load was assessed by the Anticholinergic Risk Scale (ARS), and functional recovery was assessed by the Functional Independence Measure (FIM). The primary outcome was cognitive FIM (FIM-C) gain, but we also assessed the interaction of other putative factors identified from univariate analysis. Multivariate analyses were performed, adjusting for confounding factors. RESULTS AND DISCUSSION: We included 418 participants (171 males, 247 females) with a median age of 78 years (interquartile range, 72-84 years). Multiple regression analysis revealed that ARS change, length of stay, and epilepsy were independently and negatively correlated with cognitive FIM gain. Multiple logistic regression analysis indicated that the "Comprehension" and "Memory" items of the cognitive FIM gain were independently and negatively associated with anticholinergic load. WHAT IS NEW AND CONCLUSION: A causal relationship cannot be established, but increased ARS scores during hospitalization may predict limited cognitive functional improvement in geriatric patients after stroke. Alternatively, cognitive impairment may lead to increased use of anticholinergic drugs.
Assuntos
Atividades Cotidianas/psicologia , Antagonistas Colinérgicos/administração & dosagem , Cognição/efeitos dos fármacos , Acidente Vascular Cerebral/terapia , Idoso , Idoso de 80 Anos ou mais , Antagonistas Colinérgicos/efeitos adversos , Cognição/fisiologia , Estudos de Coortes , Feminino , Hospitalização , Humanos , Tempo de Internação , Masculino , Análise Multivariada , Recuperação de Função Fisiológica/efeitos dos fármacos , Análise de Regressão , Estudos Retrospectivos , Acidente Vascular Cerebral/psicologia , Reabilitação do Acidente Vascular Cerebral/métodos , Resultado do TratamentoRESUMO
It has been reported that losartan, an angiotensin II receptor blocker, alters the circadian rhythm of melatonin secretion and significantly reduces melatonin production. However, this finding has been confirmed at the animal experiment level only, and there are no reports of studies in humans. Therefore, we performed this study to confirm the reproducibility of the aforementioned findings of animal experiments in humans. Ten male subjects who were in good general health and free from any medical condition were recruited for this study. After a preliminary observation period of 7 days, the subjects received oral losartan treatment, 50 mg daily for 7 days. Blood samplings for measurement of the plasma melatonin concentrations were performed on day 7 of the preliminary observation period and day 7 of the losartan treatment period. The circadian rhythm of melatonin secretion after the 7-day treatment with losartan showed no significant difference from that recorded before the losartan administration. The significant decrease of the home blood pressure was observed on the afternoons. The blood samples showed significant decrease of the serum sodium and uric acid levels, along with a significant increase of the serum potassium level. The pharmacological actions of losartan at the ordinarily used clinical dose level were confirmed in humans, however, no significant inhibitory effect of the drug on melatonin secretion could be confirmed. These results are expected to be useful for guiding the proper use of angiotensin II receptor blockers.
Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Ritmo Circadiano/efeitos dos fármacos , Losartan/farmacologia , Melatonina/metabolismo , Adulto , Contagem de Células Sanguíneas , Pressão Sanguínea/efeitos dos fármacos , Feminino , Humanos , Masculino , Sódio/sangue , Ácido Úrico/sangue , Adulto JovemRESUMO
Although the MHC class II 'u' haplotype is strongly associated with type 1 diabetes (T1D) in rats, the role of MHC class II in the development of tissue-specific autoimmune diseases including T1D and autoimmune thyroiditis remains unclear. To clarify this, we produced a congenic strain carrying MHC class II 'a' and 'u' haplotypes on the Komeda diabetes-prone (KDP) genetic background. The u/u homozygous animals developed T1D similar to the original KDP rat; a/u heterozygous animals did develop T1D but with delayed onset and low frequency. In contrast, none of the a/a homozygous animals developed T1D; about half of the animals with a/u heterozygous or a/a homozygous genotypes showed autoimmune thyroiditis. To investigate the role of genetic background in the development of thyroiditis, we also produced a congenic strain carrying Cblb mutation of the KDP rat on the PVG.R23 genetic background (MHC class II 'a' haplotype). The congenic rats with homozygous Cblb mutation showed autoimmune thyroiditis without T1D and slight to severe alopecia, a clinical symptom of hypothyroidism such as Hashimoto's thyroiditis. These data indicate that MHC class II is involved in the tissue-specific development of autoimmune diseases, including T1D and thyroiditis.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Tireoidite Autoimune/imunologia , Animais , Diabetes Mellitus Tipo 1/genética , Mutação , Ratos , Tireoidite Autoimune/genética , Tireoidite Autoimune/patologiaRESUMO
The information on the stability of medications is important to secure their quality. There is, however, little information about the stability of medications which assume to be kept by patients and customers. We previously showed that a delay in drug release occurs in some over-the-counter (OTC) drugs following storage in a high temperature, high humidity environment. In this study we prepared model tablet formulations containing an active ingredient and excipients to investigate the cause of this delayed release. The results reveal that delayed release occurs in preparations compounded with acetaminophen (AA) as the active ingredient and erythritol (ET) and crospovidone (CP) as excipients. In addition, ET deliquesces in a high humidity environment, then incorporates other particles during room temperature storage to form an aggregate. SEM observations and micropore distribution measurements conducted on OTC tablets that exhibit delayed release revealed that the number of intraparticle pores decreased after storage under high temperature, high humidity conditions. Thus, the delayed release by these pharmaceutical product formulations may be due to a change in the micropore structure both on the surface and within the particles, thereby decreasing the solvent infiltration pathways leading to the interior of the preparation.
Assuntos
Acetaminofen/análise , Analgésicos não Narcóticos/análise , Química Farmacêutica , Preparações de Ação Retardada , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Excipientes , Umidade , Microscopia Eletrônica de Varredura , Microscopia de Polarização , Medicamentos sem Prescrição/análise , Solubilidade , Comprimidos , TemperaturaRESUMO
Food contains components that may either increase or decrease the bioavailability of a drug. In particular, it is known that grapefruit juice and St. John's Wort induce drug interactions via an effect on the drug-metabolizing enzyme cytochrome P450 (CYP). However, interactions with membrane transporters, such as P-glycoprotein and multidrug resistance-related protein (MRP), may also influence drug bioavailability. The objective of the present study was to investigate the effects of kaempferol, a flavonoid present in food, on the cytotoxicity of anticancer drugs and the mechanisms of drug resistance in the human glioblastoma cell line T98G. Acute exposure to kaempferol inhibited the efflux of calcein, a substrate of MRP; however, chronic exposure caused no apparent effect on calcein efflux. The cytotoxicity of doxorubicin was not influenced by chronic exposure of cells to kaempferol, although that of cisplatin was significantly reduced. Multidrug resistance is often associated with increased levels of MRP1, glutathione S-transferase (GST) and activity by chronic exposure to kaempferol, although MRP2 protein levels are decreased. Accordingly, we hypothesized that the cytotoxicity of anticancer drugs that conjugate with glutathione and the substrate of MRPs may be influenced by long-term intake of drugs such as kaempferol, which are substrates of MRPs and GST.
Assuntos
Antioxidantes/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Quempferóis/farmacologia , Subfamília B de Transportador de Cassetes de Ligação de ATP/biossíntese , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos/farmacologia , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fluoresceínas/farmacologia , Interações Alimento-Droga , Glutationa Transferase/biossíntese , Glutationa Transferase/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Humanos , Proteína 2 Associada à Farmacorresistência Múltipla , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVE: Sivelestat sodium hydrate (sivelestat) is a specific synthetic inhibitor of neutrophil elastase (NE). Various studies suggest that sivelestat treatment reduces inflammation. In this study, we tested the hypothesis that sivelestat acts as an inhibitor of inflammatory mediators and prevents nuclear factor-kB (NF-kB) activation. METHODS: In the presence and absence of sivelestat, the mouse macrophage cell line RAW 264.7 was stimulated with lipopolysaccharide (LPS) and the levels of inflammatory mediators (TNF-alpha, IL-6 and high mobility group box 1 (HMGB1)) and nitrite in the cell supernatant were measured, along with inducible nitric oxide synthase (iNOS) expression. RESULTS: While LPS administration increased the secretion of inflammatory mediators and nitric oxide (NO), sivelestat decreased the secretion of these mediators. Cell signaling studies demonstrated that sivelestat decreased NF-kB activation by inhibiting IkB phosphorylation. CONCLUSION: Sivelestat may inhibit the various inflammatory mediators through NF-kB inhibition.
Assuntos
Glicina/análogos & derivados , Elastase de Leucócito/antagonistas & inibidores , Macrófagos/metabolismo , NF-kappa B/antagonistas & inibidores , Inibidores de Serina Proteinase/metabolismo , Sulfonamidas/metabolismo , Animais , Linhagem Celular , Citocinas/metabolismo , Glicina/metabolismo , Proteína HMGB1/genética , Proteína HMGB1/metabolismo , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Nitritos/sangue , Transdução de Sinais/fisiologiaRESUMO
Although a modulatory role has been reported for α-lipoic acid (LA) on T-type Ca2+ channels in the nervous system, the acute effects of LA in vivo, particularly on nociceptive transmission in the trigeminal system, remain to be determined. The aim of the present study was to investigate whether acute intravenous LA administration to rats attenuates the excitability of wide dynamic range (WDR) spinal trigeminal nucleus caudalis (SpVc) neurons in response to nociceptive and non-nociceptive mechanical stimulation in vivo. Extracellular single unit recordings were made from seventeen SpVc neurons in response to orofacial mechanical stimulation of pentobarbital-anesthetized rats. Responses to both non-noxious and noxious mechanical stimuli were analyzed in the present study. The mean firing frequency of SpVc WDR neurons in response to both non-noxious and noxious mechanical stimuli was significantly and dose-dependently inhibited by LA (1-100â¯mM, i.v.) and maximum inhibition of the discharge frequency of both non-noxious and noxious mechanical stimuli was seen within 5â¯min. These inhibitory effects lasted for approximately 10â¯min. These results suggest that acute intravenous LA administration suppresses trigeminal sensory transmission, including nociception, via possibly blocking T-type Ca2+ channels. LA may be used as a therapeutic agent for the treatment of trigeminal nociceptive pain.
Assuntos
Nociceptividade/efeitos dos fármacos , Nociceptores/efeitos dos fármacos , Ácido Tióctico/farmacologia , Núcleo Espinal do Trigêmeo/efeitos dos fármacos , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Administração Intravenosa , Animais , Eletrofisiologia , Face/inervação , Masculino , Dor Nociceptiva/tratamento farmacológico , Dor Nociceptiva/patologia , Nociceptores/patologia , Nociceptores/fisiologia , Estimulação Física , Ratos Wistar , Pele/inervação , Núcleo Espinal do Trigêmeo/citologia , Núcleo Espinal do Trigêmeo/patologiaRESUMO
Patients with end-stage renal disease are at a high risk for cardiovascular diseases. It is controversial whether end-stage renal disease patients with low cardiac function can safely accept kidney transplant. Here, we present a 42-year-old kidney transplant recipient with severe mitral regurgitation accompanied by low cardiac function. He wanted to undergo a pre-emptive kidney transplant from his uncle. We decided to perform living kidney transplant prior to cardiac surgery. Despite adequate ultrafiltration and hemodiafiltration before operation, the patient's ejection fraction still remained 35% 1 day before transplant. He showed complete recovery of cardiac function in only 2 days after pre-emptive kidney transplant, although his body weight did not change before and after the operation. Early removal of the uremic toxin or inflammatory cytokines may play a role in rapid improvement of the cardiac function. Increase of vasoactive substances by improvement of kidney function may lead to reduction of afterload and amelioration of cardiac microcirculation. This report also suggests that optimal timing for operation might be important.
Assuntos
Falência Renal Crônica/complicações , Falência Renal Crônica/cirurgia , Transplante de Rim , Insuficiência da Valva Mitral/complicações , Adulto , Humanos , MasculinoRESUMO
BACKGROUND AND OBJECTIVE: We studied the effects of honeybee products on the in vitro formation of calcium phosphate precipitates. MATERIAL AND METHODS: Screening tests of the in vitro formation of calcium phosphate precipitates using 20 types of honey and four types of propolis were carried out using the pH drop method. RESULTS: The inhibitory effect on the rate of amorphous calcium phosphate transformation to hydroxyapatite and on the induction time varied greatly among the 20 types of honey and four types of propolis. We classified them according to their effects on decreasing the rate of amorphous calcium phosphate transformation to hydroxyapatite and/or increasing the induction time. Two of the 20 honeys showed little or no inhibition, either on the rate of amorphous calcium phosphate transformation to hydroxyapatite or on the induction time. Six of the honeys reduced the rate of amorphous calcium phosphate transformation to hydroxyapatite by 12-35% and with a 2.5- to 4.4-fold increase in the induction time. The remaining 12 honeys showed even greater activity. Because four of these 12 honeys had an inhibitory effect on the rate of amorphous calcium phosphate formation, they were excluded as candidates for anticalculus agents. Furthermore, three of the four types of propolis showed an inhibitory effect that was the same as or greater than 1-hydroxyethylidene- 1,1-bisphosphonate. CONCLUSION: These results suggest that eight honeys and three types of propolis may have potential as anticalculus agents in toothpastes and mouthwashes.
Assuntos
Abelhas , Fosfatos de Cálcio/química , Mel , Própole/química , Animais , Cálcio/química , Precipitação Química , Durapatita/química , Ácido Etidrônico/química , Frutose/química , Glucose/química , Humanos , Concentração de Íons de Hidrogênio , Maltose/química , Teste de Materiais , Fosfatos/química , Sacarose/química , Fatores de TempoRESUMO
AIMS: Anatomic resection, i.e., systematic removal of a liver segment confined by portal branches, is theoretically effective in eradicating intrahepatic metastasis of hepatocellular carcinoma (HCC). The procedure may reduce tumour recurrence and enhance survival of HCC patients. To determine the significance of anatomic resection for HCC patients, we retrospectively conducted a comparative analysis between anatomic (AR) and non-anatomic liver resection (NAR) in 113 Japanese HCC patients with a solitary tumour, a tumour located within one segment, absence or invasion of distal to second order branches of the portal vein, and absence or invasion of peripheral branches of the hepatic vein. METHODS: Patients were divided into two groups, AR group (n = 49) and NAR group (n = 64). RESULTS: The prevalence of liver damage Grade B in the NAR group was significantly greater than in the AR group (p < 0.05). Tumour-free and overall survival following liver resection was not significantly different between AR and NAR groups. In the NAR group, tumour-free and overall survival in patients with tumour exposure at the surgical margin was significantly lower than with a surgical margin greater than 0 mm (not exposed) (p < 0.05). Survival between the AR and NAR groups without tumour exposure at the surgical margin was similar. CONCLUSIONS: Anatomic resection is the theoretical aim. In HCC patients with impaired liver functions, limited liver resection without tumour exposure may provide longer tumour-free and overall survival.
Assuntos
Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/cirurgia , Hepatectomia/métodos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/cirurgia , Idoso , Ascite/epidemiologia , Carcinoma Hepatocelular/patologia , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
Pancreatic ductal adenocarcinoma (PDAC) is one of the most fatal types of cancer and the 5-year survival rate is only 5%. Several studies have suggested that cancer stem cells (CSCs) are thought to be involved in recurrence and metastasis and so it is essential to establish an approach targeting CSCs. Here we have demonstrated that cyclic guanosine monophosphate (cGMP) suppressed CD44 expression and the properties of CSCs in PDAC. Microarray analysis suggested that cGMP inhibited Forkhead box O3 (FOXO3), which is known as a tumor suppressor. Surprisingly, our data demonstrated that FOXO3 is essential for CD44 expression and the properties of CSCs. Our data also indicated that patients with high FOXO3 activation signatures had poor prognoses. This evidence suggested that cGMP induction and FOXO3 inhibition could be ideal candidates for pancreatic CSC.
Assuntos
Adenocarcinoma/genética , Biomarcadores Tumorais/genética , Carcinoma Ductal Pancreático/genética , Proteína Forkhead Box O3/genética , Receptores de Hialuronatos/genética , Adenocarcinoma/patologia , Animais , Biomarcadores Tumorais/biossíntese , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , GMP Cíclico/metabolismo , Proteína Forkhead Box O3/biossíntese , Regulação Neoplásica da Expressão Gênica , Humanos , Receptores de Hialuronatos/biossíntese , Camundongos , Análise em Microsséries , Metástase Neoplásica , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Prognóstico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In this paper, we report on the preparation of an ionic polymer bead-supported lipid system several hundred micrometers in diameter. The electrostatic attractive interactions between anionic lipids and cationic polymer beads served as a "molecular glue" to immobilize the lipids on the beads, and then the immobilized lipids prompted the spontaneous formation of lipid bilayer membranes. Confocal fluorescence microscopic techniques revealed that the lipid bilayer membranes were located along the outline of the beads. In addition, the integrity of the lipid bilayer membranes was microscopically confirmed by a low-molecular-weight dye (trypan blue) exclusion test.
Assuntos
Bicamadas Lipídicas/química , Citoesqueleto/química , Indicadores e Reagentes , Lipossomos/química , Lipídeos de Membrana/química , Proteínas de Membrana/química , Microscopia Confocal , Modelos Moleculares , Conformação Molecular , Organelas/fisiologia , Azul Tripano/químicaRESUMO
Brain mitochondrial carrier protein-1 (BMCP1), a new member of the mitochondrial uncoupling carrier, has been shown to be expressed predominantly in the brain of the mice and humans. We cloned rat BMCP1 cDNA and investigated its mRNA level during postnatal development and under various metabolic conditions. The nucleotide sequence of the cDNA revealed that rat BMCP1 protein was composed of 322 amino acid residues, and was 99 and 96% identical to the mouse and human proteins and 29, 33 and 35% identical to rat uncoupling protein (UCP) 1, UCP2 and UCP3, respectively. The molecular weight was predicted to be 36017 Da and the protein of this size was detectable when the cDNA was expressed in vitro. Using Northern blot analysis, the corresponding mRNA, approximately 1.8-kb in size, was found expressed predominantly in the cerebrum, cerebellum and hypothalamus. A unique developmental pattern was identified in the brain, where BMCP1 expression was low in their fetal life, but significantly elevated in the first postnatal week. Thereafter BMCP1 mRNA was maintained to be gradually increased. In 48-h fasted or insulin-induced hypoglycemic rats, BMCP1 mRNA expression in the hypothalamus slightly, but significantly, decreased compared with that in their appropriate controls. The present results indicate that BMCP1 may be involved in pathogenesis of mitochondrial dysfunction in neurons induced by aging or neurodegenerative disorders, and perhaps in energy balance in the brain.
Assuntos
Proteínas de Transporte/genética , Crescimento , Proteínas do Tecido Nervoso/genética , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos , Encéfalo/metabolismo , Proteínas de Transporte/química , Clonagem Molecular , DNA Complementar/química , Diabetes Mellitus Experimental/metabolismo , Feto , Regulação da Expressão Gênica , Masculino , Proteínas de Membrana Transportadoras , Proteínas de Transporte da Membrana Mitocondrial , Proteínas Mitocondriais , Proteínas de Desacoplamento Mitocondrial , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , RNA Mensageiro/análise , Ratos , Ratos Wistar , Alinhamento de Sequência , Regulação para CimaRESUMO
We isolated rat UCP2 cDNA, which has been proposed to play an important role in mammalian thermogenesis and body weight regulation. The nucleotide sequence of the cDNA revealed that the rat UCP2 protein is composed of 309 amino acid residues, and is 99% and 95% identical to the mouse and human proteins, respectively. The molecular weight of rat UCP2, calculated from the predicted amino acid sequence, was 33,369, and the UCP2 protein of this size was detected when the cDNA was expressed in vitro. Northern blot analysis revealed that the corresponding mRNA is approximately 1.7 kb in size, and is expressed in a variety of rat organs, with predominant expression in the heart, lung and spleen. UCP2 mRNA levels in the heart, liver, muscle and epididymal adipose tissue of Zucker fatty (fa/fa) rats were comparable to those in the lean littermates, while ob mRNA level markedly increased in the epididymal adipose tissue of Zucker (fa/fa) rats.
Assuntos
Proteínas de Membrana Transportadoras , Proteínas Mitocondriais , Obesidade/metabolismo , Biossíntese de Proteínas , Transcrição Gênica , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar , Humanos , Canais Iônicos , Masculino , Camundongos , Dados de Sequência Molecular , Obesidade/genética , Especificidade de Órgãos , Reação em Cadeia da Polimerase , Proteínas/química , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Ratos Zucker , Proteínas Recombinantes/biossíntese , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Proteína Desacopladora 2RESUMO
To provide tissue-specific and developmental characteristics of gene expression of rat heart uncoupling protein-2 (UCP2), we investigated developmental alterations of UCPs mRNA expression in the heart and brown adipose tissue (BAT), and examined possible up-regulators of heart UCP2 expression using in vitro studies. Heart UCP2 mRNA expression was low during the early postnatal days followed by a rapid and significant increase in the 2nd postnatal week. Heart UCP3 mRNA remained undetectable until the 2nd postnatal week when the expression reached a small but significant peak. BAT UCP1 mRNA was abundantly expressed in the neonate, but the expression rapidly decreased to the adult level. The studies using cultured cardiomyocytes demonstrated that both 10(-8) M triiodothyronine and 10(-7) M isoproterenol, but not phenylephrine, increased UCP2 mRNA expression. These results indicate that the sympathetic nervous system and/or thyroid hormones may be involved in the up-regulation of heart UCP2 gene expression during postnatal development. The increase in postnatal heart UCP2 may provide a key link between the postnatal energy shift and adaptation of rat pups to their novel environment.
Assuntos
Proteínas de Membrana Transportadoras , Proteínas Mitocondriais , Miocárdio/metabolismo , Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sistema Nervoso Simpático/metabolismo , Tri-Iodotironina/farmacologia , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/crescimento & desenvolvimento , Tecido Adiposo Marrom/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Animais , Animais Recém-Nascidos , Sequência de Bases , Células Cultivadas , Primers do DNA/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Coração/efeitos dos fármacos , Coração/crescimento & desenvolvimento , Canais Iônicos , Isoproterenol/farmacologia , Fenilefrina/farmacologia , Ratos , Ratos Wistar , Proteína Desacopladora 2 , Regulação para Cima/efeitos dos fármacosRESUMO
A family of uncoupling proteins (UCPs), free fatty acid anion transporters, plays a crucial role in energy homeostatic thermoregulation. Tumor necrosis factor-alpha (TNF-alpha), a member of the cytokine family, is well known as an endogenous pyrogen. To evaluate the interaction of TNF-alpha with UCPs in thermogenesis, effects of TNF-alpha on rat UCP gene expression were examined in intrascapular brown adipose tissue (BAT), epididymal white adipose tissue (WAT) and soleus muscle (Muscle). Administration of TNF-alpha elevated rectal temperature by 0.7 degree C as well as serum leptin which peaked at 6 h, compared with saline controls. BAT UCP1 mRNA expression was increased by 1.2-fold at 6 h after the TNF-alpha treatment and decreased by 0.8-fold at 16 h after the treatment. In contrast to UCP1 expression in BAT, UCP2 mRNA expressions in BAT, WAT, and Muscle was increased to reach maximum levels of 1.3-, 1.6- and 1.8-fold, respectively, at 16 h after the treatment. UCP3 mRNA in Muscle, but not in BAT or WAT, was exclusively up-regulated by 1.7-fold at 16 h after the treatment. These results indicate that TNF-alpha up-regulates UCP gene expression differentially and tissue dependently, and add novel insights into thermogenesis under conditions of malignancy and inflammation.
Assuntos
Proteínas de Transporte/genética , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras , Proteínas Mitocondriais , Proteínas/genética , Fator de Necrose Tumoral alfa/farmacologia , Desacopladores/metabolismo , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Animais , Sequência de Bases , Regulação da Temperatura Corporal/efeitos dos fármacos , Sondas de DNA/genética , Canais Iônicos , Leptina , Masculino , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Proteínas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Distribuição Tecidual , Proteína Desacopladora 1 , Proteína Desacopladora 2 , Regulação para Cima/efeitos dos fármacosRESUMO
Diabetic rats have a deficiency in their heart ATP concentrations, and although the mechanism remains to be elucidated, this deficiency may involve increased uncoupling of oxidative phosphorylation. To investigate whether heart uncoupling proteins (UCPs) are subject to transcriptional regulation in diabetes, we examined changes in UCP mRNA expression in the heart of streptozotocin-induced diabetic (STZ-DM) rats. Heart UCP3 mRNA expression significantly increased by 9.4-fold in STZ-DM rats, while levels of UCP2 mRNA expression were not significantly altered. Insulin supplementation in STZ-DM rats returned UCP3 mRNA concentrations to control levels. The expression of UCP3 mRNA was similarly elevated in the heart of fasted rats, which also have hypoinsulinemia and hyper-free fatty acidemia but, unlike the STZ-DM rats, are hypoglycemic. Since hyperinsulinemia alone was previously reported to not affect UCP3 gene expression in the muscle, these results indicate that hyper-free fatty acidemia is a potent enhancer of UCP3 gene expression in the diabetic rat heart. Interestingly, we found no changes in UCP3 mRNA levels in Zucker fatty (fa/fa) rats with excessive chronic hyper-free fatty acidemia, which suggests that upregulation of heart UCP3 mRNA may depend on an acute change in free fatty acid concentrations rather than on their sustained elevation. High-energy ATP deficiencies in the diabetic rat heart may primarily result from proton leakage due to the upregulation of UCP3 expression.