RESUMO
Renovascular hypertension (RHV) is the cause of high blood pressure due to left renal ischemia, and obesity and hypertension cause an inflammatory response. This work analyzed the inflammatory and tissue repair profile in renal, hepatic, and cardiac tissues in an animal model of RVH associated with a high-fat diet and caloric restriction. The expressions of RORγ-t, IL-17, T-bet, and TNF-α decreased and IFN-γ increased in the right kidney. In relation to the left kidney, caloric restriction decreased the expression of IFN-γ. In the liver, caloric restriction decreased RORγ-t, IL-17, and T-bet. Hypertension associated with obesity decreased the expression of IFN-γ, while caloric restriction increased. In the right kidney, hypertension and obesity, associated or not with caloric restriction, increased the area of collagen fibers. In the heart and liver, caloric restriction reduced the area of collagen fibers. Caloric restriction increased vascular endothelial growth factor, reduced levels of growth transformation factor-ß1 (TGF-ß), and increased collagen I in the left kidney. Hypertension/obesity, submitted or not having caloric restriction, increased TGF-ß in liver. The results suggest that caloric restriction has beneficial effects in lowering blood pressure and regulating tissue proinflammatory cytokines. However, there was no change in the structure and composition of tissue repair markers.
Assuntos
Hipertensão Renovascular , Ratos , Animais , Hipertensão Renovascular/metabolismo , Ratos Wistar , Interleucina-17 , Restrição Calórica , Fator A de Crescimento do Endotélio Vascular , Obesidade/complicações , Fator de Crescimento Transformador beta , Inflamação , Colágeno/metabolismoRESUMO
OBJECTIVE: Herein, we evaluated pinealectomy-induced melatonin absence to determine its effects on craniofacial and dental development in the offspring. DESIGN: Female Wistar rats in three groups, i.e., intact pregnant rats, pinealectomized pregnant rats (PINX), and pinealectomized pregnant rats subjected to oral melatonin replacement therapy, were crossed 30 days after surgery. The heads of 7-day-old pups were harvested for cephalometric and histological analyses, and maxillae and incisors were collected for mRNA expression analysis. RESULTS: The PINX pups exhibited a reduction in neurocranial and facial parameters such as a decrease in alveolar bone area, incisor size and proliferation, and an increase in odontoblasts and the dentin layer. Based on incisor mRNA expression analysis, we found that Dmp1 expression was upregulated, whereas Col1a1 expression was downregulated. Maxillary mRNA expression revealed that Rankl expression was upregulated, whereas that of Opn and Osx was downregulated. CONCLUSION: Our results demonstrated that the absence of maternal melatonin during early life could affect dental and maxillary development in offspring, as well as delay odontogenesis and osteogenesis in maxillary tissues. CLINICAL RELEVANCE: Our findings suggest that disruptions or a lack of melatonin during pregnancy may cause changes in craniofacial and dental development, at least in animal experiments; however, in humans, these feedings are still poorly understood, and thus careful evaluations of melatonin levels in humans need to be investigated in craniofacial alterations.
Assuntos
Melatonina , Glândula Pineal , Gravidez , Humanos , Ratos , Animais , Feminino , Melatonina/farmacologia , Melatonina/metabolismo , Ratos Wistar , Glândula Pineal/metabolismo , Glândula Pineal/cirurgia , RNA MensageiroRESUMO
Nutritional restriction during developmental periods impairs organ physiology. Female rats were subjected to protein restriction during pregnancy and lactation to analyze dental and maxillary development. Four exposure groups were considered: normal-protein diet during pregnancy and lactation (NP, 17% casein), low-protein diet during lactation (LP-L, 6% casein), low-protein diet during pregnancy and lactation (LP), and low-protein diet during pregnancy (LP-G). Maxillae from 15-day-old male pups were collected. All protein-restricted groups presented increased dentin thickness and reduced alveolar bone area. When protein restriction was applied during both gestation and lactation (LP), harmful effects were observed in the form of loss of protective OPG (osteoprotegerin) in tooth epithelium-mesenchyme, due to higher RANKL expression, delay in odontoblast maturation, less dental pulp vascularity, reduction in amount of alveolar bone, and less matrix mineralization. In the LP-L group, effects of protein restriction seemed less harmful, and despite less alveolar bone, the enhancement in BMP-7, VEGF, and RANKL seems a compensatory signal to maintain maxillary osteogenesis. In LP-G animals, Dspp expression was higher, suggesting a delay in odontoblast maturation or expression recuperation. In conclusion, maternal protein restriction affects dental and maxillary development. A low-protein diet only in gestation allows for normal development. A low-protein diet during gestation-lactation results in impaired odontogenesis that may increase susceptibility of dental anomalies.
Assuntos
Caseínas , Feminino , Masculino , Ratos , AnimaisRESUMO
BACKGROUND: Calorie restriction (CR) is a type of dietary intervention that is essential in weight loss through modulation of critical metabolic control pathways, is well established and understood in cases of systemic arterial hypertension, however, its role in renovascular hypertension is still unclear. METHODS: Rats were divided into three groups: SHAM, and two groups that underwent surgery to clip the left renal artery and induce renovascular hypertension (OH and OHR). The SHAM diet was as follows: 14 weeks normolipidic diet; OH: 2 weeks normolipidic diet + 12 weeks hyperlipidic diet, both ad libitum; OHR, 2 weeks normolipidic diet + 8 weeks ad libitum high-fat diet + 4 weeks 40% calorie-restricted high-fat diet. RESULTS: Rats in the OHR group had decreased blood pressure, body weight, and glucose levels. Reductions in insulinemia and in lipid and islet fibrotic areas in the OHR group were observed, along with increased insulin sensitivity and normalization of insulin-degrading enzyme levels. The expression of nicotinamide phosphoribosyltransferase (NAMPT), insulin receptor (IR), sirtuin 1 (SIRT1), and complex II proteins were increased in the liver tissue of the OHR group. Strong correlations, whether positive or negative, were evaluated via Spearman's model between SIRT1, AMPK, NAMPT, PGC-1α, and NNMT expressions with the restoration of normal blood pressure, weight loss, glycemic and lipid panel, and mitochondrial adaptation. CONCLUSION: CR provided short-term beneficial effects to recover the physiological parameters induced by a high-fat diet and renal artery stenosis in obese and hypertensive animals. These benefits, even in the short term, can provide physiological benefits in the long term.
Assuntos
Hipertensão Renovascular , Hipertensão , Estado Pré-Diabético , Obstrução da Artéria Renal , Animais , Restrição Calórica , Dieta Hiperlipídica , Lipídeos , Ratos , Sirtuína 1/metabolismo , Redução de PesoRESUMO
The literature has shown the beneficial effects of microcurrent (MC) therapy on tissue repair. We investigated if the application of MC at 10 µA/90 s could modulate the expression of remodeling genes transforming growth factor beta (Tgfb), connective tissue growth factor (Ctgf), insulin-like growth factor 1 (Igf1), tenascin C (Tnc), Fibronectin (Fn1), Scleraxis (Scx), Fibromodulin (Fmod) and tenomodulin in NIH/3T3 fibroblasts in a wound healing assay. The cell migration was analyzed between days 0 and 4 in both fibroblasts (F) and fibroblasts + MC (F+MC) groups. On the 4th day, cell viability and gene expression were also analyzed after daily MC application. Higher expression of Ctgf and lower expression of Tnc and Fmod, respectively, were observed in the F+MC group in relation to F group (p < 0.05), and no difference was observed between the groups for the genes Tgfb, Fn1 and Scx. In cell migration, a higher number of cells in the scratch region was observed in group F+MC (p < 0.05) compared to group F on the 4th day, and the cell viability assay showed no difference between the groups. In conclusion, MC therapy at an intensity/time of 10 µA/90 s with 4 daily applications did not affect cell viability, stimulated fibroblasts migration with the involvement of Ctgf, and reduced the Tnc and Fmod expression.
Assuntos
Fator de Crescimento do Tecido Conjuntivo/genética , Terapia por Estimulação Elétrica , Fibromodulina/genética , Tenascina/genética , Cicatrização/efeitos da radiação , Animais , Movimento Celular/efeitos da radiação , Fibronectinas/genética , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Fator de Crescimento Insulin-Like I/genética , Camundongos , Células NIH 3T3 , Fator de Crescimento Transformador beta1/genética , Cicatrização/genéticaRESUMO
Nicotinamide adenine dinucleotide (NAD)+ precursors such as nicotinamide activate sirtuins and enhance energy metabolism. The aim of this study was to evaluate the metabolic effects of nicotinamide in ovariectomized (OVX) female rats to establish molecular targets against obesity, which support the safe therapeutic application of nicotinamide. The OVX animals were divided into groups: SHAM (simulated surgery), SHAMn (two weeks of 35 mg·kg-1 nicotinamide per day, by gavage), OVX, and OVXn (two weeks of 35 mg·kg-1 nicotinamide per day, by gavage). The results indicated that nicotinamide favored lipolysis, as evidenced by an increase in free fatty acid and hepatic triglyceride levels, which were not fully normalized during the treatment period. The lipolysis appeared to be due to increased SIRT1 and mitochondrial oxidative phosphorylation in muscle and adipose tissue. There were decreases in muscle and fat nicotinamide N-methyltransferase (NNMT), which were associated with decreases in mass and triglyceride, low-density lipoprotein cholesterol (LDLc), and total cholesterol content. Nicotinamide appeared to be beneficial for the glycemic profile, with normal hepatic glycogen storage and a tendency towards insulin sensitivity in the OVXn. In the SHAMn group, nicotinamide led to glucose intolerance, together with reduced muscle expressions of nicotinamide phosphoribosyltransferase (NAMPT) and SIRT3, suggesting that there were no short-term benefits. Supplementation with nicotinamide led to tissue-specific adaptive lipid and molecular changes in OVX rats.
Assuntos
Niacinamida/farmacologia , Ovariectomia , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Metabolismo Energético/efeitos dos fármacos , Feminino , Fígado/efeitos dos fármacos , Ratos , Sirtuína 1/metabolismoRESUMO
Connexins (Cx) are essential for cardiovascular regulation and maintenance of cardio-renal response involving the natriuretic peptide family. Changes in the expression of connexins promote intercellular communication dysfunction and may induce hypertension, atherosclerosis, and several other vascular diseases. This study analyzed the expression of the genes involved in the renin-angiotensin system (RAS) and the relation of the connexins gene expression with the renovascular hypertension 2K1C in different tissues. The insertion of a silver clip induced renovascular hypertension 2K1C into the left renal artery. Biochemical measurements were made using commercial kits. Gene expression was evaluated in the liver, heart, and kidneys by RT-PCR. The genes investigated were LDLr, Hmgcr, Agt, Ren, Ace, Agtr1a, Anp, Bnp, Npr1, Cx26, Cx32, Cx37, Cx40 and Cx43. All genes involved in the RAS presented increased transcriptional levels in the 2K1C group, except hepatic Agt. The natriuretic peptides (Anp; Bnp) and the receptor genes (Npr1) appeared to increase in the heart, however, Npr1 decreased in the kidneys. In hepatic tissue, hypertension promoted increased expression of Cx32, Cx37, and Cx40 genes however, Cx26 and Cx43 genes were not influenced. Expression was upregulated for Cx37 and Cx43 in cardiac tissue in the 2K1C group, but Cx40 did not demonstrate any difference between groups. The stenotic kidney showed an upregulated expression for Cx37 vs Sham and contralateral kidney, although Cx40 and Cx43 were downregulated. Hypertension did not modify the transcriptional expression of Cx26 and Cx32. Therefore, this study indicated that RAS and cardiac response were regulated transcriptionally by renovascular hypertension 2K1C. Moreover, the results of connexin gene expression demonstrated differential transcriptional regulation in different tissues studied and suggest a relationship between cardiac and renal physiological changes as an adaptive mechanism to the hypertensive state.
RESUMO
The objective of this study was to evaluate the effects of red Light Emiting Diode (red LED) irradiation on fibroblasts in adipose-derived mesenchymal stem cells (ASC) co-culture on the scratch assay. We hypothesized that red LED irradiation could stimulate paracrine secretion of ASC, contributing to the activation of genes and molecules involved in cell migration and tissue repair. ASC were co-cultured with NIH/3T3 fibroblasts through direct contact and subjected to red LED irradiation (1.45 J/cm2/5min6s) after the scratch assay, during 4 days. Four groups were established: fibroblasts (F), fibroblasts + LED (FL), fibroblasts + ASC (FC) and fibroblasts + LED + ASC (FLC). The analyzes were based on Ctgf and Reck expression, quantification of collagen types I and III, tenomodulin, VEGF, TGF-ß1, MMP-2 and MMP-9, as well as viability analysis and cell migration. Higher Ctgf expression was observed in FC compared to F. Group FC presented higher amount of tenomodulin and VEGF in relation to the other groups. In the cell migration analysis, a higher number of cells was observed in the scratched area of the FC group on the 4th day. There were no differences between groups considering cell viability, Reck expression, amount of collagen types I and III, MMP-2 and TGF-ß1, whereas TGF-ß1 was not detected in the FC group and the MMP-9 in none of the groups. Our hypothesis was not supported by the results because the red LED irradiation decreased the healing response of ASC. An inhibitory effect of the LED irradiation associated with ASC co-culture was observed with reduction of the amount of TGF-ß1, VEGF and tenomodulin, possibly involved in the reduced cell migration. In turn, the ASC alone seem to have modulated fibroblast behavior by increasing Ctgf, VEGF and tenomodulin, leading to greater cell migration. In conclusion, red LED and ASC therapy can have independent effects on fibroblast wound healing, but the combination of both does not have a synergistic effect. Therefore, future studies with other parameters of red LED associated with ASC should be tested aiming clinical application for tissue repair.
RESUMO
Epithelial-to-mesenchymal transition (EMT) and endothelial-to-mesenchymal transition are processes that can occur under different biological conditions, including tissue healing due to hypertension and oxidative stress. The purpose of the present study was to evaluate the differences in gene expression of epithelial/endothelial and mesenchymal markers in different tissues. A two-kidney, one-clip (2K1C) renovascular hypertension rat model was used. Hypertension was induced by the clipping of the left renal artery; the rats were randomized into sham and 2K1C groups and monitored for up to 4 weeks. The gene expressions of E-cadherin (E-cad), N-cadherin (N-cad), α-smooth muscle actin (α-SMA), collagen I (COL1A1), collagen III (COL3A1) and hepatocyte growth factor (HGF) were determined by reverse transcription-PCR. The levels of the cytokines transforming growth factor-ß1, tumor necrosis factor-α, interleukin (IL)-4, IL-6 and IL-10 were evaluated using ELISAs. The levels of thiobarbituric acid reactive substances and thiol groups were measured to evaluate oxidative stress. All analyses were performed on the liver, heart and kidneys tissues of sham and model rats. The 2K1C animals exhibited a higher systolic blood pressure, as well as cardiac hypertrophy and atrophy of the left kidney. Fibrotic alterations in the heart and kidneys were observed, as was an increase in the collagen fiber areas, and higher levels of inflammatory cytokines, which are associated with the increased expression of fibroproliferative and anti-fibrotic genes. Renovascular hypertension regulated epithelial/endothelial and mesenchymal markers, including E-cad, N-cad, α-SMA and COL1A1 in the kidneys and heart. EMT in the kidneys was mediated by an increased level of inflammatory and profibrotic cytokines, as well as by oxidative stress. The data in the present study suggested that the expression of epithelial/endothelial and mesenchymal markers are differentially regulated by hypertension in the liver, heart and kidneys.
Assuntos
Antígenos de Diferenciação/biossíntese , Hipertensão Renal/metabolismo , Ativação Transcricional , Animais , Hipertensão Renal/patologia , Masculino , Especificidade de Órgãos , Ratos , Ratos WistarRESUMO
Vitamin E and caloric restriction have antioxidant effects in mammals. The aim of this study was to evaluate effects of vitamin E supplementation and caloric restriction upon insulin secretion and glucose homeostasis in rats. Male Wistar rats were distributed among the following groups: C, control group fed ad libitum; R, food quantity reduction of 40%; CV, control group supplemented with vitamin E [30 mg·kg-1·day-1]; and RV, food-restricted group supplemented with vitamin E. The experiments ran for 21 days. Glucose tolerance and insulin sensitivity was higher in the CV, R, and RV groups. Insulin secretion stimulated with different glucose concentrations was lower in the R and RV groups, compared with C and CV. In the presence of glucose and secretagogues, insulin secretion was higher in the CV group and was lower in the R and RV groups. An increase in insulin receptor occurred in the fat pad and muscle tissue of groups CV, R, and RV. Levels of hepatic insulin receptor and phospho-Akt protein were higher in groups R and RV, compared with C and CV, while muscle phospho-Akt was increased in the CV group. There was a reduction in hepatic RNA levels of the hepatocyte growth factor gene and insulin degrading enzyme in the R group, and increased levels of insulin degrading enzyme in the CV and RV groups. Thus, vitamin E supplementation and caloric restriction modulate insulin secretion by different mechanisms to maintain glucose homeostasis.
Assuntos
Glicemia/metabolismo , Homeostase/efeitos dos fármacos , Ilhotas Pancreáticas/metabolismo , Vitamina E/farmacologia , Animais , Antioxidantes/metabolismo , Homeostase/fisiologia , Resistência à Insulina/fisiologia , Secreção de Insulina/efeitos dos fármacos , Secreção de Insulina/fisiologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Ratos WistarRESUMO
The aim of this study was to evaluate markers of bone loss and immune response present in evolution of periodontal disease. One hundred and two Wistar rats were divided into three animals groups: PD0, without ligation and PD15 days and PD60 days, submitted to ligation placement with a sterile 3-0 silk cord in the cervical region of the upper first molar on both sides. Samples were obtained from the gingival tissue for histomorphometric analysis, immunohistochemical analysis of RANK, RANKL, OPG, characterization of the inflammatory infiltrate, quantification of nitric oxide, MCP-1, RANTES, IP10 chemokines, and expression of the TGF-b1, VEG, and bFGF. The number of inflammatory cells in gingival tissue was higher in PD60 samples. The collagen content and the area occupied by birefringent collagen fibers were lower for PD60. Differential leukocyte counting showed that there was a significantly higher polymorphonuclear influx in group PD15, while PD60 showed a greater number of lymphocytes. PD60 showed higher RANTES, IP-10, MCP-1 gene transcripts, as well as a higher nitric oxide concentration. Clinical evaluation revealed that the PD60 group presented an increase in furcal area. In conclusion, in this animal model the increase of RANK/RANKL and HGF markers is related to a specific immune response, and probably contributed to the evolution of periodontal disease. Investigating the effect of these biomarkers can help in targeted therapy for bone resorption, since blocking these can inhibit bone loss.
Assuntos
Osteoprotegerina/metabolismo , Doenças Periodontais/imunologia , Ligante RANK/metabolismo , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Animais , Western Blotting , Quimiocinas/metabolismo , Imuno-Histoquímica , Inflamação/metabolismo , Masculino , Doenças Periodontais/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Abstract The aim of this study was to evaluate markers of bone loss and immune response present in evolution of periodontal disease. One hundred and two Wistar rats were divided into three animals groups: PD0, without ligation and PD15 days and PD60 days, submitted to ligation placement with a sterile 3-0 silk cord in the cervical region of the upper first molar on both sides. Samples were obtained from the gingival tissue for histomorphometric analysis, immunohistochemical analysis of RANK, RANKL, OPG, characterization of the inflammatory infiltrate, quantification of nitric oxide, MCP-1, RANTES, IP10 chemokines, and expression of the TGF-b1, VEG, and bFGF. The number of inflammatory cells in gingival tissue was higher in PD60 samples. The collagen content and the area occupied by birefringent collagen fibers were lower for PD60. Differential leukocyte counting showed that there was a significantly higher polymorphonuclear influx in group PD15, while PD60 showed a greater number of lymphocytes. PD60 showed higher RANTES, IP-10, MCP-1 gene transcripts, as well as a higher nitric oxide concentration. Clinical evaluation revealed that the PD60 group presented an increase in furcal area. In conclusion, in this animal model the increase of RANK/RANKL and HGF markers is related to a specific immune response, and probably contributed to the evolution of periodontal disease. Investigating the effect of these biomarkers can help in targeted therapy for bone resorption, since blocking these can inhibit bone loss.
Resumo Este estudo avaliou marcadores de perda óssea e da resposta imune presentes na evolução da doença periodontal. Cento e dois ratos Wistar foram divididos em três grupos de animais: PD0, sem ligadura e PD15 dias e PD60 dias, submetidos a colocação de ligadura com um fio de seda estéril 3-0 na região cervical do primeiro molar superior em ambos os lados. Foram obtidas amostras de tecido gengival para análise histomorfométrica, análises imunohistoquímicas de RANK, RANKL, OPG, caracterização do infiltrado inflamatório, quantificação de óxido nítrico, expressão de quimiocinas MCP-1, RANTES, IP10 e do TGF-b1, VEGF e bFGF . O número de células inflamatórias no tecido gengival foi maior nas amostras PD60. O teor de colágeno na área ocupada pelas fibras de colágeno birrefringentes foram menores para PD60. A contagem diferencial de leucócitos mostrou que houve um influxo polimorfonuclear significativamente maior no grupo PD15, enquanto que PD60 mostrou número maior de linfócitos. PD60 apresentou transcritos de genes RANTES, IP-10, MCP-1 mais elevados, bem como uma maior concentração de óxido nítrico. A avaliação clínica revelou que o grupo PD60 apresentou aumento da área óssea exposta na região da furca. Em conclusão, neste modelo animal o aumento dos marcadores RANK/RANKL e HGF está relacionado a uma resposta imunológica específica e provavelmente contribuiu para a evolução da doença periodontal. Investigar o efeito destes biomarcadores pode ajudar na terapia dirigida para a reabsorção óssea, uma vez que bloquear estes pode inibir a perda óssea.
Assuntos
Animais , Masculino , Ratos , Doenças Periodontais/imunologia , Ligante RANK/metabolismo , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Osteoprotegerina/metabolismo , Doenças Periodontais/metabolismo , Imuno-Histoquímica , Western Blotting , Ratos Wistar , Quimiocinas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inflamação/metabolismoRESUMO
Background: The dietary limitation during pregnancy influences the growth and development of the fetus and offspring and their health into adult life. The mechanisms underlying the adverse effects of gestational protein restriction (GPR) in the development of the offspring hearts are not well understood. Objectives: The aim of this study was to evaluate the effects of GPR on cardiac structure in male rat offspring at day 60 after birth (d60). Methods: Pregnant Wistar rats were fed a normal-protein (NP, 17% casein) or low-protein (LP, 6% casein) diet. Blood pressure (BP) values from 60-day-old male offspring were measured by an indirect tail-cuff method using an electro sphygmomanometer. Hearts (d60) were collected for assessment of connexin 43 (Cx43) mRNA expression and morphological and morphometric analysis. Results: LP offspring showed no difference in body weight, although they were born lighter than NP offspring. BP levels were significantly higher in the LP group. We observed a significant increase in the area occupied by collagen fibers, a decrease in the number of cardiomyocytes by 104 µm2, and an increase in cardiomyocyte area associated with an increased Cx43 expression. Conclusion: GPR changes myocardial levels of Cx43 mRNA in male young adult rats, suggesting that this mechanism aims to compensate the fibrotic process by the accumulation of collagen fibers in the heart interstitium.
Fundamento: A limitação dietética durante a gravidez influencia o crescimento e desenvolvimento do feto e da prole e sua saúde na vida adulta. Os mecanismos subjacentes dos efeitos adversos da restrição proteica gestacional (RPG) no desenvolvimento dos corações da prole não são bem compreendidos. Objetivos: Avaliar os efeitos da RPG sobre a estrutura cardíaca em filhotes machos de ratas aos 60 dias após o nascimento (d60). Métodos: Ratos fêmeas Wistar grávidas foram alimentadas com uma dieta de proteína normal (PN, 17% caseína) ou de baixa proteína (BP, caseína 6%). Os valores de pressão arterial (PA) de descendentes do sexo masculino de 60 dias de idade foram medidos por meio de um método indireto de manguito de cauda usando um eletro esfigmomanômetro. Os corações (d60) foram coletados para avaliação da expressão de RNAm da conexina 43 (Cx43) e análise morfológica e morfométrica. Resultados: A prole BP não mostrou diferença no peso corporal, embora tenha nascido mais leve do que a prole PN. Os níveis de PA foram significativamente mais altos no grupo BP. Observou-se um aumento significativo na área ocupada pelas fibras colágenas, diminuição do número de cardiomiócitos em 104 µm2 e aumento da área de cardiomiócitos associada ao aumento da expressão de Cx43. Conclusão: A RPG altera os níveis miocárdicos de RNAm de Cx43 em ratos adultos jovens, sugerindo que este mecanismo visa compensar o processo fibrótico pelo acúmulo de fibras de colágeno no interstício cardíaco.
Assuntos
Conexina 43/metabolismo , Dieta com Restrição de Proteínas , Miocárdio/metabolismo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , RNA Mensageiro/metabolismo , Animais , Conexina 43/análise , Feminino , Humanos , Masculino , Gravidez , RNA Mensageiro/análise , Ratos , Ratos WistarRESUMO
Abstract Background: The dietary limitation during pregnancy influences the growth and development of the fetus and offspring and their health into adult life. The mechanisms underlying the adverse effects of gestational protein restriction (GPR) in the development of the offspring hearts are not well understood. Objectives: The aim of this study was to evaluate the effects of GPR on cardiac structure in male rat offspring at day 60 after birth (d60). Methods: Pregnant Wistar rats were fed a normal-protein (NP, 17% casein) or low-protein (LP, 6% casein) diet. Blood pressure (BP) values from 60-day-old male offspring were measured by an indirect tail-cuff method using an electro sphygmomanometer. Hearts (d60) were collected for assessment of connexin 43 (Cx43) mRNA expression and morphological and morphometric analysis. Results: LP offspring showed no difference in body weight, although they were born lighter than NP offspring. BP levels were significantly higher in the LP group. We observed a significant increase in the area occupied by collagen fibers, a decrease in the number of cardiomyocytes by 104 µm2, and an increase in cardiomyocyte area associated with an increased Cx43 expression. Conclusion: GPR changes myocardial levels of Cx43 mRNA in male young adult rats, suggesting that this mechanism aims to compensate the fibrotic process by the accumulation of collagen fibers in the heart interstitium.
Resumo Fundamento: A limitação dietética durante a gravidez influencia o crescimento e desenvolvimento do feto e da prole e sua saúde na vida adulta. Os mecanismos subjacentes dos efeitos adversos da restrição proteica gestacional (RPG) no desenvolvimento dos corações da prole não são bem compreendidos. Objetivos: Avaliar os efeitos da RPG sobre a estrutura cardíaca em filhotes machos de ratas aos 60 dias após o nascimento (d60). Métodos: Ratos fêmeas Wistar grávidas foram alimentadas com uma dieta de proteína normal (PN, 17% caseína) ou de baixa proteína (BP, caseína 6%). Os valores de pressão arterial (PA) de descendentes do sexo masculino de 60 dias de idade foram medidos por meio de um método indireto de manguito de cauda usando um eletro esfigmomanômetro. Os corações (d60) foram coletados para avaliação da expressão de RNAm da conexina 43 (Cx43) e análise morfológica e morfométrica. Resultados: A prole BP não mostrou diferença no peso corporal, embora tenha nascido mais leve do que a prole PN. Os níveis de PA foram significativamente mais altos no grupo BP. Observou-se um aumento significativo na área ocupada pelas fibras colágenas, diminuição do número de cardiomiócitos em 104 µm2 e aumento da área de cardiomiócitos associada ao aumento da expressão de Cx43. Conclusão: A RPG altera os níveis miocárdicos de RNAm de Cx43 em ratos adultos jovens, sugerindo que este mecanismo visa compensar o processo fibrótico pelo acúmulo de fibras de colágeno no interstício cardíaco.
Assuntos
Humanos , Animais , Masculino , Feminino , Gravidez , Ratos , Efeitos Tardios da Exposição Pré-Natal/metabolismo , RNA Mensageiro/metabolismo , Conexina 43/metabolismo , Dieta com Restrição de Proteínas , Miocárdio/metabolismo , RNA Mensageiro/análise , Ratos Wistar , Conexina 43/análiseRESUMO
BACKGROUND: Human and animal evidence suggests that environmental toxicants may have an adverse impact on male reproductive health, reducing the population's reproductive output. Owing to the renewed attraction for natural products, some of them constitute effective alternatives to mitigate these effects. Propolis is a candidate for this use because of its intrinsic properties. In many situations, it improved the testicular damage and alleviated the toxic effects induced by environmental contaminant exposure. OBJECTIVE: The aim of this study was to investigate possible alterations of testicular parameters and certify if its use is really advantageous to the testis, since this could affect rat reproductive function. MATERIALS AND METHODS: Forty-eight adult male Wistar rats were divided into four groups (Co = control, T1 = 3 mg propolis/kg/day, T2 = 6 mg/kg/day, T3 = 10 mg/kg/day) and were exposed during 56 days. The testes were assessed with morphometrical, stereological, and ultrastructural analyses. Cell proliferation and death were diagnosed, respectively, by immunocytochemistry. Connexin 43 (Cx43) and N-cadherin transcript levels were determined by reverse transcription-polymerase chain reaction. RESULTS: Increased cell proliferation and Leydig cell volume were observed in T2, and in contrast, Cx43 upregulation and cell death were observed in T3. Both T2 and T3 showed ultrastructural abnormalities in testicular parenchyma. CONCLUSION: We recommend a cautious intake of propolis to avoid deleterious effects. SUMMARY: Chronic intake of Brazilian green propolis induced N.-cadherin downregulation and decreased on seminiferous tubule volumeIncrease on connexin 43 expression and cell death and decrease in Leydig cell.(LC) number/testis with the concentration of 10 mg/kg/day were observedIncrease on cell proliferation, cytoplasmic proportion, and volume of LC with the concentration of 6 mg/kg/day was detectedThe presence of empty spaces between spermatids and malformed spermatozoa in the lumen of seminiferous tubule was showedThis male reproductive disruption can be linked to phenolic compounds present in Brazilian green propolis. Abbreviation Used: AEC: 3-amino-9-ethylcarbazole; AJ: Adherens junction; AME: Aromadendrin-40-methyl ether; CAPE: Caffeic acid phenethyl ester; Co: Control group; C×43: Connexin 43; DAB: Diaminobenzidine; dNTP: Deoxyribonucleotide phosphate; DSP: Daily sperm production; FA: Ferulic acid; FSH: Follicle-stimulating hormone; GJ: Gap junction; GJIC: Gap junction intercellular communication; HPLC: High-performance liquid chromatography; LC: Leydig cell; LH: Luteinizing hormone; N-cad: N-cadherin; PCNA: Proliferating cell nuclear antigen; PCR: Polymerase chain reaction; RT-PCR: Reverse transcription-polymerase chain reaction; SDM: Standard deviation of mean; T1: Group exposed to 3 mg of propolis/kg/day; T2: Group exposed to 6 mg of propolis/kg/day; T3: Group exposed to 10 mg of propolis/kg/day; TUNEL: Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; WB-ras 2 cells: Ras-transformed rat liver epithelial cell line.
RESUMO
OBJECTIVE: Complexes like conjugated linoleic acid (CLA) reduce the percentage of body fat by increasing energy expenditure, fat oxidation, or both. The aim of this study was to verify if CLA is able to mimic caloric restriction (CR), and determine the effects of CLA on liver metabolic profile of young adult male Wistar rats. MATERIALS AND METHODS: We divided 36 animals into the following groups: 1) Control; 2) CLA (1% of daily food intake, 21 days, orogastric intubation); 3) Restr (fed 60% of the diet offered to controls); and 4) CLA Restr. Liver tissues were processed for biochemical and molecular or mitochondrial isolation (differential centrifugation) and blood samples were collected for biochemical analyses. RESULTS: Treatment of the animals for 21 days with 1% CLA alone or combined with CR increased liver weight and respiration rates of liver mitochondria suggesting significant mitochondrial uncoupling. We observed a decrease in adipose tissue leading to insulin resistance, hyperinsulinemia, and hepatic steatosis due to increased liver cholesterol and triacylglycerol levels, but no significant effects on body mass. The expression of hepatic cellular connexins (43 and 26) was significantly higher in the CLA group compared with the Control or Restr groups. CONCLUSION: CLA does not seem to be a safe compound to induce mass loss because it upregulates the mRNA expression of connexins and induces hepatic mitochondrial changes and lipids disorders.
Assuntos
Restrição Calórica , Metabolismo Energético , Fígado Gorduroso/prevenção & controle , Ácidos Linoleicos Conjugados/administração & dosagem , Fígado/metabolismo , Animais , Metabolismo dos Lipídeos , Masculino , Ratos , Ratos Wistar , Fatores de TempoRESUMO
ABSTRACT Objective Complexes like conjugated linoleic acid (CLA) reduce the percentage of body fat by increasing energy expenditure, fat oxidation, or both. The aim of this study was to verify if CLA is able to mimic caloric restriction (CR), and determine the effects of CLA on liver metabolic profile of young adult male Wistar rats. Materials and methods We divided 36 animals into the following groups: 1) Control; 2) CLA (1% of daily food intake, 21 days, orogastric intubation); 3) Restr (fed 60% of the diet offered to controls); and 4) CLA Restr. Liver tissues were processed for biochemical and molecular or mitochondrial isolation (differential centrifugation) and blood samples were collected for biochemical analyses. Results Treatment of the animals for 21 days with 1% CLA alone or combined with CR increased liver weight and respiration rates of liver mitochondria suggesting significant mitochondrial uncoupling. We observed a decrease in adipose tissue leading to insulin resistance, hyperinsulinemia, and hepatic steatosis due to increased liver cholesterol and triacylglycerol levels, but no significant effects on body mass. The expression of hepatic cellular connexins (43 and 26) was significantly higher in the CLA group compared with the Control or Restr groups. Conclusion CLA does not seem to be a safe compound to induce mass loss because it upregulates the mRNA expression of connexins and induces hepatic mitochondrial changes and lipids disorders.
Assuntos
Animais , Masculino , Ratos , Restrição Calórica , Ácidos Linoleicos Conjugados/administração & dosagem , Metabolismo Energético , Fígado Gorduroso/prevenção & controle , Fígado/metabolismo , Fatores de Tempo , Ratos Wistar , Metabolismo dos LipídeosRESUMO
Connexins (Cx) and cadherins are responsible for cell homeostasis. The Cx activity is directly related to cholesterol. The present work investigates whether vitamin E, with or without caloric restriction (CR), alters the mRNA expression of Cx26, Cx32, Cx43, N-cadherins (N-cads), E-cadherins (E-cads) and alpha-smooth muscle actin (α-SMA), and evaluates their relation to cholesterol metabolism in rat liver. Animals were divided into different groups: control with ad libitum diet (C), control+vitamin E (CV), aloric restriction with intake to 60% of group C (CR), and the intake of group CR+vitamin E (RV). There were increases of manganese superoxide dismutase (Mn-SOD) and glutathione S-transferase mu 1, indicating antioxidant effects of CR and vitamin E. An increase of nitric oxide in the CR group was in agreement with the Mn-SOD data. Supplementation with vitamin E, with or without CR, upregulated the expression of Cx26 mRNA and increased low-density lipoprotein cholesterol (LDL-c) in the CV group. Reductions of Cx32 and Cx43 were associated with lower LDL-c. Increases in Hmgcr and low-density lipoprotein receptor (LDLr) in the CV and RV groups could be explained by the effect of vitamin E. A reduction of LDLr in the CR group was due to the reduced dietary intake. Increases in cadherins in the CV, CR and RV groups were indicative of tissue maintenance, which was also supported by increases of α-SMA in groups CV and RV. Finally, vitamin E, with or without CR, increased Cx26, probably modulated by expression of the Hmgcr and LDLr genes. This suggests important relationship of Cxs and cholesterol metabolism genes.
Assuntos
Caderinas/metabolismo , Restrição Calórica , Conexinas/metabolismo , Fígado/efeitos dos fármacos , Proteínas do Tecido Nervoso/metabolismo , Vitamina E/farmacologia , Animais , Caderinas/genética , LDL-Colesterol/sangue , Conexina 26 , Conexina 43/genética , Conexina 43/metabolismo , Conexinas/genética , Dieta , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Homeostase , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Masculino , Proteínas do Tecido Nervoso/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Proteína beta-1 de Junções ComunicantesRESUMO
ABSTRACT Objective: To evaluate whether the single nucleotide polymorphism rs7895833 (A/G) of the gene SIRT1 is associated with metabolic syndrome criteria in a sample of Brazilian adults. Methods: Serum samples and oral mucosal cells were collected from 243 subjects aged 30 to 70 years. Biochemical, hormonal, and anthropometric data were obtained. The single nucleotide polymorphism rs7895833 (A/G) was analyzed by polymerase chain reaction using the amplification refractory mutation system. Results: Among the 243 study subjects, 100 (41.15%) were classified as non-metabolic syndrome and 143 (58.85%), as metabolic syndrome. The frequency of the single nucleotide polymorphism rs7895833 (A/G) did not differ between the groups. However, 111 patients (45.67%) were overweight (body mass index: 25-29.9 kg/m2). Blood glucose, total cholesterol, triglycerides, very low density lipoprotein, low density lipoprotein, waist and hip circumferences, and blood pressure were higher in the metabolic syndrome group than in the non-metabolic syndrome group. Free thyroxine 4, grown hormone, and insulin levels were within the normal range. The metabolic conditions of the patients with metabolic syndrome indicate biochemical, anthropometric, and hormonal changes characteristic of overweight and obesity. Conclusion: The SIRT1 polymorphism rs7895833 (A/G) is not associated with the metabolic syndrome in the adult Brazilian population.
RESUMO Objetivo: Avaliar se o polimorfismo de nucleotídeo único rs7895833 (A/G) do gene SIRT1 está associado à síndrome metabólica em uma amostra da população brasileira. Foram coletadas, de 243 indivíduos com idades entre 30 e 70 anos, amostras de soro e de células da mucosa bucal. Métodos: Dosagens bioquímicas, hormonais e dados antropométricos foram analisados. O polimorfismo de nucleotídeo único rs7895833 (A/G) foi analisado por sistema de amplificação de mutação por refração - reação em cadeia da polimerase. Resultados: Entre os 243 indivíduos estudados, 100 (41,15%) foram classificados como não apresentando síndrome meta-bólica e 143 (58,85%) como apresentando a síndrome. Não houve diferença significativa na frequência do polimorfismo de nucleotídeo único rs7895833 (A/G) entre os grupos. No entanto, 111 pacientes (45,67%) estavam com sobrepeso (índice de massa corporal: 25-29,9 kg/m2). Glicose, colesterol total, triglicerídeos, lipoproteínas de muito baixa densidade, lipoproteínas de baixa densidade, circunferência da cintura e do quadril e pressão arterial foram maiores no grupo com síndrome metabólica quando comparado ao outro grupo. Tiroxina 4 livre, hormônio do crescimento e os níveis de insulina estavam no valor de referência. As condições metabólicas dos pacientes com síndrome metabólica indicam alterações bioquímicas, antropométricas e hormonais características do excesso de peso e da obesidade. Conclusão: Sugerimos que o polimorfismo rs7895833 (A/G), no gene SIRT1, não esteja associado à síndrome metabólica na população adulta brasileira.
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Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Índice de Massa Corporal , Polimorfismo de Nucleotídeo Único , Síndrome Metabólica , Sirtuína 1 , ObesidadeRESUMO
Single nucleotide polymorphisms (SNPs) are important markers in many studies that link DNA sequence variations to phenotypic changes; such studies are expected to advance the understanding of human physiology and elucidate the molecular basis of diseases. The DFNB1 locus, which contains the GJB2 and GJB6 genes, plays a key role in nonsyndromic hearing loss. Previous studies have identified important mutations in this locus, but the contribution of SNPs in the genes has not yet been much investigated. The aim of this study was to investigate the association of nine polymorphisms located within the DFNB1 locus with the occurrence of autosomal recessive nonsyndromic hearing loss (ARNSHL). The SNPs rs3751385 (C/T), rs7994748 (C/T), rs7329857 (C/T), rs7987302 (G/A), rs7322538 (G/A), rs9315400 (C/T), rs877098 (C/T), rs945369 (A/C), and rs7333214 (T/G) were genotyped in 122 deaf patients and 132 healthy controls using allele-specific PCR. There were statistically significant differences between patients and controls, in terms of allelic frequencies in the SNPs rs3751385, rs7994748, rs7329857, rs7987302, rs945369, and rs7333214 (P < 0.05). No significant differences between the two groups were observed for rs7322538, rs9315400, and rs877098. Our results suggest that SNPs present in the GJB2 and GJB6 genes may have an influence on ARNSHL in humans.