Effects of Low Protein Diet on Nuclear Factor Erythroid 2-Related Factor 2 Gene Expression in Nondialysis Chronic Kidney Disease Patients.

OBJECTIVE(S): Low protein diets (LPD; 0.6 g/kg/day), prescribed for nondialysis chronic kidney disease (CKD) patients, have demonstrated numerous benefits. LPDs may modulate inflammation and oxidative stress through the nuclear factor erythroid 2-related factor 2 (Nrf2), which encodes antioxidant and phase II detoxifying enzymes. LPDs also inhibit or antagonize nuclear factor kB (NF-kB) activity, which orchestrates inflammatory and oxidative stress responses. The objective of this study was to evaluate the effects of LPD on Nfr2 and NF-κB messenger RNA (mRNA) expression in nondialysis CKD patients. METHODS: In this longitudinal study, a LPD was prescribed for 30 nondialysis CKD patients for 6 months. Peripheral blood mononuclear cells were isolated, and quantitative real-time polymerase chain reaction analysis was performed to evaluate Nrf2, NF-κB, and NADPH quinine oxidoreductase-1 mRNA expression. Thiobarbituric acid-reactive substance (TBARS) levels, a marker of lipid peroxidation, were also evaluated. RESULTS: (Age 55.5 ± 14.0 years; body mass index 29.1 ± 5.9 kg/m2; glomerular filtration rate 35.6 ± 12.2 mL/minute). After 6 months of nutritional intervention, Nrf2 mRNA expression increased from 0.85 (0.47-1.56) to 1.28 (0.63-2.63) nmol/mL (P = .03), and TBARS levels were significantly decreased from 1.78 (1.31-2.38) to 1.30 (1.07-2.22) nmol/mL (P = .04). NF-κB mRNA expression showed no significant difference after 6 months, but the Nrf2/NF-κB ratio was increased. CONCLUSION(S): In this study, a LPD appeared to modulate Nrf2 expression and decrease the levels of TBARS in nondialysis CKD patients. However, more studies are needed to confirm the effectiveness of LPD on the modulation of transcription factors involved with oxidative stress and inflammation in nondialysis CKD patients.

Nrf2, NF-κB and PPARβ/δ mRNA Expression Profile in Patients with Coronary Artery Disease / Perfil da Expressão do mRNA do Nrf2, NF-κB e PPARβ/δ em Pacientes com Doença Arterial Coronariana

Abstract Background: Oxidative stress and inflammation are present in coronary artery disease (CAD) and are linked to the activation of the transcription nuclear factor kappa B (NF-κB). To attenuate these complications, transcription factors like nuclear factor erythroid 2-related factor 2 (Nrf2) and peroxisome proliferator-activated receptor-β/δ (PPARβ/δ) can be activated to inhibit NF-κB. However, the available data on expression of NF-κB, Nrf2 and PPARβ/δ in CAD patients are limited. Objective: To evaluate the expression of the transcription factors NF-κB and Nrf2 and PPAR��/�� in CAD patients. Methods: Thirty-five patients (17 men, mean age 62.4 ? 7.55 years) with CAD and twelve patients (5 men, mean age 63.50 ? 11.46 years) without CAD were enrolled. Peripheral blood mononuclear cells (PBMCs) were isolated and processed for mRNA expression of Nrf2, NF-κB, NADPH: quinone oxidoreductase 1 (NQO1) and PPARβ/δ mRNAs using quantitative real-time polymerase chain reaction (qPCR). p < 0.05 was considered statistically significant. Results: There was no difference in the mRNA expressions of Nrf2 (1.35 ? 0.57), NF-κB (1.08 ? 0.50) or in the antioxidant enzyme NQO1 (1.05 ? 0.88) in the CAD group compared to the group without CAD (1.16 ? 0.76, 0.95 ? 0.33, 0.81 ? 0.55, respectively). However, PPARβ/δ was highest expressed in the CAD group (1.17 ? 0.86 vs. 0.56 ? 0.34, p = 0.008). Conclusion: The main finding of this study was the PPARβ/δ being more expressed in the PBMC of patients with CAD compared to the control group, whereas no differences were observed in Nrf2 or NF-κB mRNA expressions.

Resumo Fundamentos: O estresse oxidativo e a inflamação estão presentes na doença arterial coronariana (DAC) e estão ligados à ativação do fator de transcrição nuclear kappa B (NF-κB). Para atenuar essas complicações, fatores de transcrição como o fator nuclear eritroide 2-relacionado ao fator 2 (Nrf2) e o receptor ativado por proliferador de peroxissoma β/δ (PPARβ/δ) podem ser ativados para inibir o NF-κB. No entanto, os dados disponíveis sobre a expressão de NF-κB, Nrf2 e PPARβ/δ em pacientes com DAC são limitados. Objetivo: Avaliar a expressão dos fatores transcricionais NF-κB e Nrf2 e o PPARβ/δ em pacientes com DAC. Métodos: Trinta e cinco pacientes (17 homens, idade média de 62,4 ± 7,55 anos) com DAC e doze pacientes (5 homens, com idade média de 63,50 ± 11,46 anos) sem DAC foram incluídos. Células mononucleares do sangue periférico (PBMCs) foram isoladas e processadas para a expressão de mRNA do Nrf2, NF-κB, NADPH: quinona oxidoredutase 1 (NQO1) e mRNAs do PPARβ/δ por meio de reação em cadeia da polimerase quantitativa em tempo real (qPCR). Valores de p < 0,05 foram considerados como estatisticamente significativos. Resultados: Não houve diferença nas expressões de mRNA do Nrf2 (1,35 ± 0,57), NF-κB (1,08 ± 0,50) ou na enzima antioxidante NQO1 (1,05 ± 0,88) no grupo DAC em comparação com o grupo sem DAC (1,16 ± 0,76, 0,95 ± 0,33, 0,81 ± 0,55, respectivamente). Entretanto, o PPARβ/δ apresentou maior expressão no grupo com DAC (1,17 ± 0,86 vs. 0,56 ± 0,34, p = 0,008). Conclusão: O principal achado do presente estudo foi o PPARβ/δ apresentar maior expressão nas PBMCs de pacientes com DAC comparados ao grupo controle, ao passo que não foram observadas diferenças nas expressões de mRNA do Nrf2 ou NF-κB.

Nrf2, NF-κB and PPARß/δ mRNA Expression Profile in Patients with Coronary Artery Disease.

BACKGROUND: Oxidative stress and inflammation are present in coronary artery disease (CAD) and are linked to the activation of the transcription nuclear factor kappa B (NF-κB). To attenuate these complications, transcription factors like nuclear factor erythroid 2-related factor 2 (Nrf2) and peroxisome proliferator-activated receptor-ß/δ (PPARß/δ) can be activated to inhibit NF-κB. However, the available data on expression of NF-κB, Nrf2 and PPARß/δ in CAD patients are limited. OBJECTIVE: To evaluate the expression of the transcription factors NF-κB and Nrf2 and PPAR𝛽/𝛿 in CAD patients. METHODS: Thirty-five patients (17 men, mean age 62.4 ? 7.55 years) with CAD and twelve patients (5 men, mean age 63.50 ? 11.46 years) without CAD were enrolled. Peripheral blood mononuclear cells (PBMCs) were isolated and processed for mRNA expression of Nrf2, NF-κB, NADPH: quinone oxidoreductase 1 (NQO1) and PPARß/δ mRNAs using quantitative real-time polymerase chain reaction (qPCR). p < 0.05 was considered statistically significant. RESULTS: There was no difference in the mRNA expressions of Nrf2 (1.35 ? 0.57), NF-κB (1.08 ? 0.50) or in the antioxidant enzyme NQO1 (1.05 ? 0.88) in the CAD group compared to the group without CAD (1.16 ? 0.76, 0.95 ? 0.33, 0.81 ? 0.55, respectively). However, PPARß/δ was highest expressed in the CAD group (1.17 ? 0.86 vs. 0.56 ? 0.34, p = 0.008). CONCLUSION: The main finding of this study was the PPARß/δ being more expressed in the PBMC of patients with CAD compared to the control group, whereas no differences were observed in Nrf2 or NF-κB mRNA expressions.

Aryl Hydrocarbon Receptor and Uremic Toxins from the Gut Microbiota in Chronic Kidney Disease Patients: Is There a Relationship between Them?

Recent studies have suggested that uremic toxins such as indoxyl sulfate (IS) and indole-3-acetic acid (IAA) from the metabolism of the gut microbiota may be involved in the inflammatory signaling pathway in chronic kidney disease (CKD) patients through the activation of the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor. The objective of this study was to investigate the possible relationship between uremic toxins (IS and IAA) and AhR protein expression in CKD patients. A cross-sectional observational study involving 17 hemodialysis (HD) [11 men, 55.5 ± 11.7 years of age, 54.0 (25.5-136.0) months of HD, body mass index (BMI) of 25.8 ± 3.8 kg/m2] and 15 non-dialysis-dependent (NDD) CKD (8 men, 54.1 ± 18.2 years of age, glomerular filtration rate of 34.8 ± 21.0 mL/min/1.73 m2, BMI of 27.4 ± 5.0 kg/m2) patients was conducted. IS and IAA levels were measured by reversed-phase high-performance liquid chromatography, and the protein expression levels of AhR and nuclear factor κ B (NF-κB) were evaluated by a Western blot assay. There was no difference in the expression of either AhR or NF-κB in the patients, and as expected, uremic toxin levels were higher in HD patients than in NDD patients. In the overall analysis, AhR protein expression was positively associated with IAA plasma levels ( r = 0.4; p = 0.03) and NF-κB protein expression ( r = 0.62; p = 0.001). Although the role of AhR in inflammation and CVD in CKD patients is far from being completely understood, the association between IAA and AhR observed in this study suggests a possible role for uremic toxins in the cell signaling pathway involved in inflammation in CKD patients.

Effects of Low-Protein Diet on lipid and anthropometric profiles of patients with chronic kidney disease on conservative management / Efeitos da dieta hipoproteica sobre os perfis lipídico e antropométrico de pacientes com doença renal crônica em tratamento conservador

ABSTRACT Introduction: Chronic Kidney disease (CKD) patients have a high prevalence of cardiovascular mortality, and among the risk factors are dyslipidemia and obesity, common findings in the early stages of CKD. The aim of this study was to evaluate the effects of low protein diet (LPD) on the lipid and anthropometric profile in non-dialysis CKD patients. Methods: Forty CKD patients were studied (20 men, 62.7 ± 15.2 years, glomerular filtration rate (GFR) 26.16 ± 9.4 mL/min/1.73m2). LPD (0.6g/kg/d) was prescribed for six months and, biochemical and anthropometric parameters like body mass index (BMI), waist circumference and body fat mass (assessed by dual X-ray absorptiometry - DXA) were evaluated before and after six months with LPD. Results: After six months of nutritional intervention, patients presented reduction on BMI (from 28.1 ± 5.6 to 27.0 ± 5.3 Kg/m2, p = 0.001), total cholesterol (from 199.7 ± 57.1 to 176.0 ± 43.6mg/dL, p = 0.0001), LDL (from 116.2 ± 48.1 to 97.4 ± 39.1 mg/dL, p = 0,001) and uric acid (from 6.8 ± 1.4 to 6.2 ± 1.3 mg/dL, p = 0.004). In addition, GFR values were increased from 26.2 ± 9.5 to 28.9 ± 12.7mL/min (p = 0.02). The energy, proteins, cholesterol and fiber intake were reduced significantly. Conclusion: LPD prescribe to non-dialysis CKD patients for six months was able to improve some cardiovascular risk factors as overweight and plasma lipid profile, suggesting that LPD can be also an important tool for protection against cardiovascular diseases in these patients.

RESUMO Introdução: Pacientes com Doença Renal Crônica (DRC) possuem alta prevalência de mortalidade cardiovascular e, dentre os fatores de risco, encontram-se alterações no perfil lipídico e excesso de peso, que são achados comuns na DRC. O objetivo deste estudo foi avaliar os efeitos da dieta hipoproteica sobre o perfil antropométrico e lipídico de pacientes com DRC em tratamento conservador. Métodos: Foram estudados 40 pacientes com DRC (20 homens, 62,7 ± 15,2 anos, e Taxa de Filtração Glomerular (TFG) de 26,2 ± 9,4 mL/min/1,73m2). Os pacientes receberam prescrição de dieta hipoproteica (0,6g/kg/d) e parâmetros bioquímicos e antropométricos como índice de massa corporal (IMC), circunferência da cintura (CC) e percentual de gordura corporal (GC) avaliado por absorciometria com raio-x de dupla energia (DXA), foram analisados antes e após 6 meses de intervenção. Resultados: Os pacientes apresentaram após 6 meses, redução do IMC (de 28,1 ± 5,6 para 27,0 ± 5,3Kg/m2, p = 0,001), colesterol total (de 199,7 ± 57,1 para 176,0 ± 43,6mg/dL, p = 0,0001), LDL (de 116,2 ± 48,1 para 97,4 ± 39,1 mg/dL, p = 0,001) e ácido úrico (de 6,8 ± 1,4 para 6,2 ± 1,3 mg/dL, p = 0,004) e, aumento da TFG de 26,2 ± 9,5 para 28,9 ± 12,7mL/min (p = 0,02). Houve redução significativa na ingestão de energia e proteínas, bem como de colesterol e fibras. Conclusão: A intervenção com dieta hipoproteica para pacientes com DRC em tratamento conservador por seis meses foi capaz de melhorar alguns fatores de risco cardiovascular, como o excesso de peso e o perfil lipídico plasmático, sugerindo que a dieta hipoproteica, além de outros benefícios pode também ser importante ferramenta para a proteção de doenças cardiovasculares nesses pacientes.

Effects of Low-Protein Diet on lipid and anthropometric profiles of patients with chronic kidney disease on conservative management.

INTRODUCTION: Chronic Kidney disease (CKD) patients have a high prevalence of cardiovascular mortality, and among the risk factors are dyslipidemia and obesity, common findings in the early stages of CKD. The aim of this study was to evaluate the effects of low protein diet (LPD) on the lipid and anthropometric profile in non-dialysis CKD patients. METHODS: Forty CKD patients were studied (20 men, 62.7 ± 15.2 years, glomerular filtration rate (GFR) 26.16 ± 9.4 mL/min/1.73m2). LPD (0.6g/kg/d) was prescribed for six months and, biochemical and anthropometric parameters like body mass index (BMI), waist circumference and body fat mass (assessed by dual X-ray absorptiometry - DXA) were evaluated before and after six months with LPD. RESULTS: After six months of nutritional intervention, patients presented reduction on BMI (from 28.1 ± 5.6 to 27.0 ± 5.3 Kg/m2, p = 0.001), total cholesterol (from 199.7 ± 57.1 to 176.0 ± 43.6mg/dL, p = 0.0001), LDL (from 116.2 ± 48.1 to 97.4 ± 39.1 mg/dL, p = 0,001) and uric acid (from 6.8 ± 1.4 to 6.2 ± 1.3 mg/dL, p = 0.004). In addition, GFR values were increased from 26.2 ± 9.5 to 28.9 ± 12.7mL/min (p = 0.02). The energy, proteins, cholesterol and fiber intake were reduced significantly. CONCLUSION: LPD prescribe to non-dialysis CKD patients for six months was able to improve some cardiovascular risk factors as overweight and plasma lipid profile, suggesting that LPD can be also an important tool for protection against cardiovascular diseases in these patients.

Could Low-Protein Diet Modulate Nrf2 Pathway in Chronic Kidney Disease?

Oxidative stress and inflammation are common findings in chronic kidney disease (CKD) patients, and they are directly linked to clinical outcomes such as protein energy wasting and cardiovascular disease. Nuclear factor-erythroid 2-related factor 2 (Nrf2) is the master regulator of antioxidant genes, regulating the expression of detoxifying enzymes of phase II and antioxidant responses. Furthermore, Nrf2 can also regulate anti-inflammatory cellular responses, by inhibiting nuclear factor kappa B activity (transcription factor that promotes inflammation). Therefore, modulating Nrf2 can be a new therapeutic approach to reduce inflammation and oxidative stress in CKD. Low-protein diet (LPD) prescribed for nondialysis CKD patients presents numerous benefits already well established, including reduction of inflammation and oxidative stress. However, there is no available data regarding the relationship between LPD and Nrf2 modulation in these patients. This review aims to discuss the impact, if any, of LPD on Nrf2 expression, in nondialysis CKD patients.

Effect of maternal use of flaxseed oil during pregnancy and lactation on glucose metabolism and pancreas histomorphometry of male offspring from diabetic rats.

AIM: Investigate if the maternal use of flaxseed oil prevents pancreatic alterations in the offspring of diabetic mothers. METHODS: Diabetes was induced in female wistar rats (n=12) by a high-fat diet and low-dose of streptozotocin. After the confirmation of the diabetes (glucose >300 mg/dL), rats were mated and once pregnancy was confirmed, they were allocated into three groups (n=6): high-fat group (HFG); flaxseed oil group (FOG); and control group (CG) (nondiabetic rats). At weaning, male offspring (n=12/group) received a standard chow diet. The animals were euthanized in two phases: at 100 and at 180 days, (n=6/group). The pancreas was collected for histomorphometric and immunohistochemistry analysis. RESULTS: HFG showed hypertrophy of pancreatic islets at 100 and at 180 days (p<0.0001), while the FOG offspring had islets with smaller diameters compared to HFG at both phases of sacrifice (p<0.0001). HFG had a lower percentage of small islets when compared to CG and FOG, which had a higher percentage when compared to HFG (p=0.0053) at 100 days. At 180 days HFG showed higher percentage of larger islets (p=0.00137) and lower percentage of smaller islets (p=0.00112), when compared to FOG. HFG showed lower islet insulin immunodensity at 100 days (p<0.0001) and 180 days (p<0.0001), whereas FOG was similar to CG (p<0.0001) at 100 days and higher at 180 days (p<0.0001). CONCLUSIONS: Flaxseed oil reduced the damage caused by maternal hyperglycemia, promoting normal pancreas histomorphometry and ß cell mass.

The use of flaxseed flour during pregnancy and lactation reverses lower birth weight in offspring from diabetic mothers but averts the development during lactation.

UNLABELLED: Diabetes is a complication which occurring during gestation might substantially influence the development of offspring during fetal life and postnatally. Flaxseed is a source of omega-3, that the appropriate supply during gestation and lactation are determinant for a suitable perinatal growth and development. The present study aimed to assess beneficial effects of the use of flaxseed flour during pregnancy and lactation on body development from birth to weaning of offspring from diabetic mothers. METHODS: twelve rats from a total of eighteen were induced to diabetes by high-fat diet during four weeks, also receiving one lower dose of streptozotocin. After confirmation of diabetes (glucose>300 mg/dL), they were mated and when pregnancy was confirmed, they were divided in 3 groups: high-fat group (HFG), high-fat flaxseed flour group (HFFFG) and control group (CG), receiving high- fat diet, high-fat diet added flaxseed flour and control diet, respectively. They were fed this way during whole gestation and lactation. The body development of offspring was measured weekly since the first day after birth until weaning. RESULTS: At birth, the average body mass of offspring from diabetics mothers who received only high-fat diet was 23,6% lighter than body mass of offspring from non-diabetics mothers (p<0,05), while the animals from diabetic mothers who consumed flaxseed flour during pregnancy and lactation showed the same body mass than the control group. During all experiment HFFFG group showed decreased body mass (about 20%, p<0,05)in comparison with control group. CONCLUSION: The treatment with flaxseed flour was capable of avoiding lower birth weight in offspring from diabetic mothers. However, the consumption of flaxseed flour by mothers during lactation yielded decrease offspring weight at weaning.

La diabetes es una complicación que ocurre durante la gestación puede influir sustancialmente el desarrollo de las crías durante la vida fetal y postnatal. La linaza es una fuente de ácidos grasos omega-3, que la oferta apropiado durante la gestación y lactancia son determinantes para un adecuados crecimiento y desarrollo perinatal. Este estudio tuvo como objetivo evaluar los efectos beneficiosos del uso de la harina de linaza durante el embarazo y la lactancia en el desarrollo corporal desde el nacimiento hasta el destete de las crías de madres diabéticas. Métodos: Los doce ratas, de un total de dieciocho fueron inducidas a la diabetes con dieta alta en grasas durante cuatro semanas también recibir una dosis reducida de estreptozotocina. Después de la confirmación de la diabetes (glucosa> 300mg/dL), que fueron apareadas y cuando se confirmó el embarazo, fueron divididos en 3 grupos: grupo de alto contenido de grasa (HFG), grupo de alto contenido de grasa con harina de linaza (HFFFG) y grupo control (GC ), recibiendo la dieta alta en grasas, dieta alta en grasa añadida harina de linaza y dieta control, respectivamente. Fueron alimentados de esta manera durante toda la gestación y la lactancia. El el desarrollo corporal de las crías se midió semanalmente desde el primer día después de su nacimiento hasta el destete. Resultados: En el nacimiento, la masa corporal medio de las crías de madres diabéticas que recibieron sólo la dieta rica en grasas era 23,6% más ligero que la masa corporal de las crías de los no diabéticos madres (p.