Experimental model of nephropathy associated with diabetes mellitus in mice

Purpose: Nontransmissible chronic diseases, such as diabetes mellitus (DM) and nephropathy, affect a significant portion of the population, often treated due to injuries that require healing and regeneration. To create an experimental model of associated comorbidities, for healing and regeneration studies, protocols for induction of nephropathy by ischemia and reperfusion (I/R) and induction of DM by injection of streptozotocin (STZ) were associated. Methods: Sixty-four mice (Mus musculus), female, adult, Swiss strain, weighing approximately 20 g, were divided into four groups: G1: control (n = 24), G2: nephropathy group (N) (n = 7), G3, DM (n = 9), and G4: N+DM (n = 24). Arteriovenous stenosis (I/R) of the left kidney was performed as the first protocol. The animals received a hyperlipidemic diet for 7 days after the injection of STZ (150 mg/kg, via i.p.) and an aqueous glucose solution (10%) for 24 h. The animals in the G3 and G4 groups were observed for 14 days before receiving the diet and STZ. The evolution of nephropathy was observed using a urine test strip and the DM, through the analysis of blood glucose with a reagent strip on a digital monitor. Results: The ischemic induction protocols of nephropathy and DM with STZ, associated, were sustainable, low-cost, and without deaths. There were alterations compatible with initial renal alterations, in the first 14 days, such as increased urinary density, pH alteration, presence of glucose, proteins and leukocytes, when compared to the control group. DM was confirmed by the presence of hyperglycemia 7 days after induction and its evolution after 14 days. The animals in the G4 group showed constant weight loss when compared to the other groups. It was possible to observe morphological alterations in the kidneys submitted to I/R, regarding coloration, during surgery and after the end of the observation period, in the volume and size of the left kidney, when compared to the contralateral kidney. Conclusion: It was possible to induce nephropathy and DM associated in the same animal, in a simple way, confirmed with rapid tests, without losses, providing a basis for future studies.

Role of Brain Angiotensin-II in Development of Experimental Diabetic Nephropathy in Wistar Rats

Abstract The renin-angiotensin-aldosterone system (RAAS) plays a key role in diabetic nephropathy (DN). Angiotensin-II secreted during the RAAS pathway increases nephropathy. It stimulates oxidative stress which can quench nitric oxide. Reduced nitric oxide level aggravates Ang-II-induced vasoconstriction. Ang-II has also emerged as a central mediator of the glomerular hemodynamic changes that are associated with renal injury. Deletion of ACE2 is also noted due to increased Ang-II level which leads to the development of DN. We hypothesize that nephropathy caused by Ang-II in the periphery may be controlled by brain RAAS. ACE inhibitors and ARBs may show the renoprotective effect when administered through ICV without crossing the blood-brain barrier. DN was observed after 8 weeks of diabetes induction through alloxan. Administration of captopril and valsartan once and in combined therapy for 2 weeks, significantly reduced urine output, blood urea nitrogen, total protein in the urine, serum cholesterol, serum creatinine, serum triglycerides, and kidney/body weight ratio as compared to diabetic control rats. Further, combination therapy significantly increased the body weight and serum nitrate level as compared to diabetic control animals. However, increased ACE2 levels in the brain may reduce the sympathetic outflow and might have decreased the peripheral activity of Ang-II which shows beneficial effects in DN.

Recomendaciones para evaluar la tasa de filtrado glomerular menor de 60 ml/min/1, 73 m2 en pacientes con enfermedad renal y diabetes mellitus / Recommendations to evaluate the glomerular filtration rate below 60 ml/min/1. 73 m2 in patients with kidney disease and diabetes

Diagnosticar, clasificar y estadificar la enfermedad renal en pacientes con diabetes mellitus (DM) es un desafío tanto para los médicos de atención primaria como para los especialistas, porque no existe método en la práctica clínica que evalúe la tasa de filtrado glomerular (TFG) en forma precisa. Para evaluar la función renal en enfermedad renal crónica (ERC) con menos de 60 ml/min/1.73m2 , correspondiente a los estadios 3, 4 y 5 de la clasificación actual, los métodos disponibles en los laboratorios clínicos son de limitada exactitud. En este trabajo se desarrollarán las condiciones que debería cumplir un marcador ideal, las dificultades que ofrece la evaluación de la creatinina, la medición de la TFG, así como las ventajas y limitaciones de las recomendaciones del uso de fórmulas para su determinación, y el algoritmo actual para estimar función renal. Conclusiones: actualmente, a pesar de las limitaciones, se recomienda el empleo de fórmulas para la estimación de la TFG, sobre todo en TFG menor a 60 ml/min/1.73 m2. Es un desafío, para un futuro mediato, desarrollar mejores recursos para su evaluación.

To diagnose,classify and stage diabetic kidney disease in patients with diabetes mellitus is a challenge in clinical practice for both primary care physicians and specialists because there is no method in clinical practice that evaluates accurately the glomerular filtration rate (GFR). This challenge is due to difficulties in evaluating kidney function in stages of chronic kidney disease (CKD) below 60 ml/min/1.73m2 corresponding to stages 3, 4 and 5 of the classification, because the available tools in clinical laboratories are of limited accuracy. This work explores the conditions that an ideal marker should meet, the difficulties offered by the evaluation of serum creatinine, the measurement of the glomerular filtration rate (GFR) as well as the advantages and limitations of the recommendations of the use of formulas for its determination and the current algorithm to estimate renal function. Conclusions: currently, despite the limitations, the use of formulas for the diagnosis of renal function is recommended, especially in GFR below 60 ml/min/1.73 m2. It is a challenge for the near future to develop better tools for the evaluation of TGF.

Acacia senegal inhibits diabetic nephropathy ultrastructure alteration and renal biomarkers changes in rats / Acacia senegal inhibe la alteración de la ultraestructura de la nefropatía diabética y los cambios en los biomarcadores renales en rata

SUMMARY: Diabetic nephropathy (DN) is the most common complication of diabetes. Several studies have been done in a trial to protect against this problem at the ultrastructure level. This study investigates the protective effect of oral administration of Acacia senegal (AS) against the development of DN. Sixty male albino rats were randomly divided into six groups: control, Acacia senegal control, Diabetic untreated, diabetic insulin-treated, Diabetic AS treated, and Diabetic insulin and AS combined treated groups. Plasma glucose, HbA1c, serum Albumin, creatinine, urine creatinine was measured using specific kits. Determinations of creatinine clearance and blood pressure were done. The renal tissues of both kidneys were prepared to investigate under both light (LM) and electron microscope (EM). Ultrastructure examination of renal rats tissue of diabetic untreated rats showed the destruction of the glomerular basement membrane and endothelial cells together with hemorrhage in glomerular capsules (Bowman's capsules). On the other side, both LM and EM revealed improving the endothelial cells and the other glomerular capsules structures, especially with the combined treated group, which confirmed the improvement of the biochemical investigation in the study. In conclusion, from the present study, using the oral AS together with SC insulin could be protected against the development of DN.

RESUMEN: La nefropatía diabética (ND) es la complicación más común de la diabetes. Se han realizado varios estudios de ensayo para abordar esta dificultad a nivel de ultraestructura. Este estudio investiga el efecto protector de la administración oral de Acacia senegal (AS) contra el desarrollo de la ND. Se dividieron sesenta ratas albinas machos aleatoriamente en seis grupos: control, control de Acacia senegal, diabéticos no tratados, diabéticos tratados con insulina, diabéticos tratados con AS y grupos tratados con compuesto de insulina diabética + AS. Se midieron utilizando kits específicos, glucosa plasmática, HbA1c, albúmina sérica, creatinina en sangre y en orina. Se registraron la creatinina y la presión arterial. Los tejidos renales de ambos riñones se prepararon para investigar tanto con microscopio óptico (MO) como electrónico (ME). El examen de la ultraestructura del tejido renal de ratas diabéticas no tratadas mostró la destrucción de la membrana basal glomerular y las células endoteliales junto con hemorragia en las cápsulas glomerulares (cápsulas de Bowman). Por otro lado, tanto MO como ME revelaron una mejora de las células endoteliales y las estructuras capsulares glomerulares, en el grupo tratado con el compuesto, lo que confirmó la mejora de la investigación bioquímica. En conclusión, el uso de AS oral en combinación con insulina podría proteger contra el desarrollo de ND.

Análisis geográfico de nefropatía diabética e insuficiencia renal en el primer nivel de atención, IMSS 2019 / Geographic analysis of diabetic nephropathy and renal insufficiency in the primary care, IMSS 2019

Introducción: en un contexto donde la prevalencia de diabetes mellitus e hipertensión arterial ha aumentado significativamente en años recientes, las enfermedades renales adquieren importancia por la potencial demanda de atención especializada y de recursos en salud que requieren. Objetivo: analizar la distribución geográfica de la nefropatía diabética (ND) y la insuficiencia renal (IR) con base en las consultas otorgadas en unidades de primer nivel del Instituto Mexicano del Seguro Social (IMSS) durante 2019, para identificar las unidades médicas con mayor carga de atención. Material y métodos: estudio ecológico-exploratorio en el que se estimaron indicadores por cada mil derechohabientes en relación a las consultas otorgadas por ND e IR según la ocasión de servicio, la unidad médica familiar (UMF) de primer nivel y la representación. Se utilizó estadística espacial para analizar dichos indicadores. Resultados: el 45% de las consultas otorgadas fue por ND y el 52.4% por IR. La mayor carga por ND se registró en la UMF No. 50 de Cd. Juárez (Chihuahua) y en la No. 49 Gabino Barreda (Veracruz Sur), con 1.7 consultas de primera vez y 148.3 subsecuentes por mil derechohabientes, respectivamente. Mientras que en la UMF No. 40 Manlio Fabio Altamirano y No. 25 Cotaxtla, en Veracruz Norte, la mayor carga fue por IR, con 4.9 consultas de primera vez y 134.2 subsecuentes por mil derechohabientes, respectivamente. Conclusiones: los resultados podrían contribuir al fortalecimiento de las unidades médicas que así lo requieran y en la distribución eficiente de los recursos disponibles para atender la demanda de servicios de salud de ND e IR en el IMSS

Background: In a context where the prevalence of Diabetes Mellitus and Hypertension has increased significantly in recent years, kidney diseases become important for the potential demand for specialized health care and resources required. Objective: To analyze the geographical distribution of Diabetic Nephropathy (DN) and Renal Insufficiency (RI) based on the medical consultations given in first-level units of IMSS during 2019, to identify the medical units with the highest burden of care. Material and methods: Ecological-exploratory study in which indicators were estimated for every thousand persons in relation to medical consultations given by ND and RI according to service time, first-level medical unit (UMF) and representation to analyze the magnitude and geographic distribution at the national level. Results: 45% of medical consultations were by ND and 52.4% by RI. The highest burden per DN was registered in UMF No. 50 Cd. Juarez (Chihuahua) and No. 49 Gabino Barreda (Veracruz Sur), with 1.7 first-time medical consultations and 148.3 subsequent medical consultations per 1,000 persons, respectively. While in UMF No. 40 Manlio Fabio Altamirano and No. 25 Cotaxtla, in Veracruz Norte, the highest burden was for RI, with 4.9 first-time medical consultations and 134.2 subsequent medical consultations per 1000 persons, respectively. Conclusions: The results could contribute to strengthening of medical units where it is necessary and the efficient allocation of resources available to meet the demand for health services of ND and RI in IMSS.

The A allele of the rs759853 single nucleotide polymorphism in the AKR1B1 gene confers risk for diabetic kidney disease in patients with type 2 diabetes from a Brazilian population

ABSTRACT Objective: The AKR1B1 gene encodes an enzyme that catalyzes the reduction of glucose into sorbitol. Chronic hyperglycemia in patients with diabetes mellitus (DM) leads to increased AKR1B1 affinity for glucose and, consequently, sorbitol accumulation. Elevated sorbitol increases oxidative stress, which is one of the main pathways related to chronic complications of diabetes, including diabetic kidney disease (DKD). Accordingly, some studies have suggested the rs759853 polymorphism in the AKR1B1 gene is associated with DKD; however, findings are still contradictory. The aim was to investigate the association of the rs759853 polymorphism in the AKR1B1 gene and DKD. Materials and methods: The sample comprised 695 patients with type 2 DM (T2DM) and DKD (cases) and 310 patients with T2DM of more than 10 years' duration, but no DKD (controls). The polymorphism was genotyped by real-time PCR. Results: Allelic and genotype frequencies of this polymorphism did not differ significantly between groups. However, the A/A genotype was associated with risk for DKD after adjustment for gender, triglycerides, BMI, presence of hypertension and diabetic retinopathy, and duration of DM, under both recessive (P = 0.048) and additive (P = 0.037) inheritance models. Conclusion: Our data suggest an association between the AKR1B1 rs759853A/A genotype and risk for DKD in Brazilians T2DM patients.

Exploring the therapeutic mechanism of Longqi Fang for diabetic kidney disease based on network pharmacology and verification in rats / 南方医科大学学报

OBJECTIVE@#To study the therapeutic mechanism of Longqi Fang (LQF) for diabetic kidney disease (DKD) based on GEO database and network pharmacology.@*METHODS@#LQF and DKD targets were obtained using the databases including GEO, TCMSP, CNKI, ChemDraw, and SwissTarget Prediction, and LQF-DKD intersection targets were obtained with VENNY. String was used for protein-protein interaction (PPI) analysis, and R package for KEGG and GO enrichment analysis. Cytoscape 3.7.2 software Network graphs were constructed. The results of network pharmacology analysis were verified in SD rat models of DKD by daily treatment of the rats with LQF at low (1 g/kg), medium (2 g/kg), and high (2 g/kg) doses, and kidney pathology was observed with HE staining and the changes in renal function were assessed. Western blotting was used to detect the expression levels of NF-κB and p-NF-κB proteins.@*RESULTS@#We identified 760 main targets of LQF, and obtained 1026 differential genes using GEO database and 61 LQF-DKD intersection targets using Venny database. The core targets obtained through PPI network analysis included Myc, EGF, CASP3, VEGFA, CCL2, SPP1, VCAM1 and ICAM1. Go analysis showed that LQF affects mainly nuclear receptor activity and ligand activated transcription factor activity. KEGG analysis showed that LQF affects inflammatory signaling pathways by interfering with NF-κB, TNF, and PI3K-AKT. In rat models of DKD, treatment with LQF resulted in significant improvements of the renal functions (P < 0.05) and glomerular and tubular structure and arrangement in a dose-dependent manner. Western blotting results showed that LQF dose-dependently downregulated NF-κB and p-NF-κB expressions in the rat models.@*CONCLUSION@#The therapeutic mechanism of LQF for DKD involves multiple components, targets and signal pathways that mediate an inhibitory effect on NF-κB signaling pathway to protect the renal function.

Mechanisms of Coreopsis tinctoria Nutt in the treatment of diabetic nephropathy based on network pharmacyology analysis of its active ingredients / Mecanismos de Coreopsis tinctoria Nutt. En el tratamiento de la nefropatía diabética basado en el análisis farmacológico de red de sus principios activos

SUMMARY: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) can protect diabetic kidneys, but the mechanisms are unclear. This work is to investigate the potential mechanisms of C. tinctoria Nutt. in the treatment of diabetic nephropathy based on network pharmacology analysis of its active ingredients. Twelve small molecular compounds of C. tinctoria Nutt. and targets related to diabetic nephropathy were docked by Discovery Studio 3.0. DAVID database was used for GO enrichment and KEGG pathway analysis. Cytoscape 3.6.1 was used to construct active ingredient-target network. Cell viability was detected with MTT. Glucose consumption was analyzed with glucose oxidase method. Protein expression was measured with Western blot and immunofluorescence. Electron microscopy observed autophagosomes. The core active ingredients of C. tinctoria Nutt. included heriguard, flavanomarein, maritimein, and marein. Twenty-one core targets of the 43 potential targets were PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK, and ABL2. These 21 core targets were significantly enriched in 50 signaling pathways. Thirty- four signaling pathways were closely related to diabetic nephropathy, of which the top pathways were PI3K/AKT, insulin, and mTOR, and insulin resistance. The enriched GO terms included biological processes of protein phosphorylation, and the positive regulation of PI3K signaling and cytokine secretion; cellular components of cytosol, extracellular region, and extracellular space; and molecular function of protein kinase activity, ATP binding, and non-membrane spanning protein tyrosine kinase activity. In vitro experiments found that marein increased the expression of phosphorylated AKT/AKT in human renal glomerular endothelial cells of an insulin resistance model induced by high glucose, as well as increased and decreased, respectively, the levels of the microtubule-associated proteins, LC3 and P62. C. tinctoria Nutt. has many active ingredients, with main ingredients of heriguard, flavanomarein, maritimein, and marein, and may exert anti-diabetic nephropathy effect through various signaling pathways and targets.

RESUMEN: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) puede proteger riñones diabéticos, sin embargo los mecanismos son desconocidos. Este trabajo se realizó para investigar los potenciales mecanismos de C. tinctoria Nutt. en el tratamiento de la nefropatía diabética basado en el análisis de farmacología en red de sus principios activos. Doce compuestos moleculares pequeños de C. tinctoria Nutt. y los objetivos relacionados con la nefropatía diabética fueron acoplados por Discovery Studio 3.0. La base de datos DAVID se utilizó para el enriquecimiento GO y el análisis de la vía KEGG. Se usó Cytoscape 3.6.1 para construir una red de ingrediente-objetivo activa. La viabili- dad celular se detectó mediante MTT. El consumo de glucosa se analizó con el método de glucosa oxidasa. La expresión proteica fue determinada mediante Western blot e inmunofluorescencia. En la microscopía electrónica se observó autofagosomas. Los principales ingredientes activos de C. tinctoria Nutt. incluyeron heriguard, flavanomarein, maritimin y marein. Veintiún de los 43 objetivos potenciales fueron PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK y ABL2. Estos 21 objetivos principales se enriquecieron significativamente en 50 vías de señalización. Treinta y cuatro vías de señalización estuvieron estrechamente relacionadas con la nefropatía diabética, de las cuales las principales vías fueron PI3K/ AKT, insulina y mTOR, y resistencia a la insulina. Los términos GO enriquecidos incluyeron procesos biológicos de fosforilación proteica, la regulación positiva de la señalización de PI3K y la secreción de citoquinas; componentes celulares del citosol, región extracelular y espacio extracelular; y la función molecular de la actividad de la proteína quinasa, la unión de ATP y la actividad de la proteína tirosina quinasa que no se extiende por la membrana. Los experimentos in vitro encontraron que la mareína aumentaba la expresión de AKT/AKT fosforilada en células endoteliales glomerulares renales humanas en un modelo de resistencia a la insulina inducida por niveles elevados de glucosa, así como aumentaron y disminuyeron respectivamente, los niveles de las proteínas asociadas a los microtúbulos, LC3 y P62. C. tinctoria Nutt. tiene muchos principios activos, con ingredientes principales de heriguard, flavanomarein, maritimain y marein, y puede ejercer un efecto de nefropatía antidiabética a través de distintass vías de señalización y objetivos.

Association of low birth weight and prematurity with blood pressure and biomarkers of renal and endocrine-pancreatic functions in adults without nephropathy or diabetes / Associação de baixo peso ao nascer com pressão arterial e biomarcadores de função renal e endócrino-pancreática em adultos sem diabetes e nefropatia

Introduction: Low birth weight (LBW), a proxy for hostile intrauterine environment, has been associated with these diseases in adulthood. In Brazil, there is scarce literature on the association of LBW with blood pressure (BP) or kidney and pancreatic functions in adults. The big ELSA-Brasil database allowed to explore whether: 1) pre-diabetic individuals could have kidney function impairment, detectable by renal biomarkers; 2) LBW is associated with less favorable BP levels and kidney and pancreatic function in adulthood compared to normal birth weight. Objectives: To analyze the association of LBW with BP and biomarkers of kidney and endocrine-pancreatic function in adults without DM or nephropathy. The specific objectives were: Paper 1: to review the literature on the prevalence of diabetic kidney disease (DKD) in pre-diabetic individuals. Paper 2: to assess serum Cystatin C (sCys C) as an early marker of kidney dysfunction in individuals without DM. Paper 3: to compare BP levels and kidney function biomarkers (estimated glomerular filtration rate - eGFR, albumin-creatinine ratio - ACR and sCys C) according to the presence of LBW and to analyze their associations with BP and kidney function biomarkers in individuals without DM or nephropathy. Paper 4: to compare markers of ß-cell function and insulin sensitivity (HOMA-ß, HOMA-IR, HOMA-AD, QUICKI, TyG and TG/HDL) according to the presence of LBW and to analyze LBW associations with markers of ß-cell function and insulin sensitivity. Methods: Cross-sectional analysis of ELSA-Brasil data includes 2 fronts: assessment of the LBW associations with BP and kidney function and with endocrine-pancreatic function. Individuals aged > 60 years, BMI < 18.5 kg/m², DM, kidney, thyroid and liver dysfunction were excluded. Sociodemographic data, lifestyle, birth weight and previous diseases were collected by questionnaires, and clinical and laboratory data in the HU/USP. Dependent variables were BP, biomarkers of kidney and pancreatic functions, and independent variable was LBW. Categorical variables were compared using the chi-squared test and continuous variables by Student t test or the Wilcoxon test. Multiple linear regression models were employed to analyze associations between LBW and the outcome variables. Directed acyclic graph (DAG) was used to make the minimum necessary adjustment to the models. The propensity score method was applied to homogenize differences in sample size. Results: Paper 1: Prevalence of DKD ranged from 4.5 to 26.0% in pre-diabetic individuals. Considering eGFR in isolation, the prevalence rates varied from 4.5 to 21.3%, based only on ACR from 7.0 to 26.0% and based on combined criteria the prevalence was between 12.3 and 17.7%. Paper 2: Pre-diabetic individuals had higher sCys C levels than normoglycemic ones [0.67 (0.41-0.95) vs 0.48 (0.31-0.81) mg/L, p<0.001] and lower eGFR (96.3±17.4 vs 100.6±17.1 mL/min/1.73m², p<0.001). Normoglycemic hyperfiltrating individuals had lower sCys C than normofiltrating ones (p=0.035). Comparing eGFR levels between groups, this gradually decreased as the sCys C and ACR parameters worsened (p-trend=0.06). Paper 3: The LBW group had higher systolic (p=0.015) and diastolic BP (p=0.014) and ACR values (p=0.031), and lower eGFR (p=0.015) than normal birth weight. The preterm group had higher mean BP levels, but no difference in kidney function was detected. In a regression model, BP levels were associated with LBW, but this association disappeared after adding prematurity, which remained associated with BP (p=0.017). Having applied propensity score matching, LBW was associated with ACR (p=0.003), but not with eGFR or BP levels. Paper 4: Individuals with LBW or normal birth weight reported similar BMI at the age 20 years and current BMI was slightly lower in the LBW group. Cardiometabolic and endocrine-pancreatic function parameters were within normal ranges. In regression analysis, log-transformed HOMA-ß, but not the other indexes, was associated with LBW (p=0.014) independent of sex, skin color, prematurity, and family history of DM. After applying propensity score matching LBW was associated with HOMA-AD and TG/HDL indexes. Discussion: Our findings suggest that individuals with near-normal glucose metabolism disturbance could have some impaired kidney function. Looking at early-life risk factors is relevant since their associations with BP and renal and pancreatic function biomarkers could already be identified even in healthy individuals, without DM and nephropathy. Prospective studies are needed to assess the predictive value aiming at proposing prevention measures.

Introdução: O baixo peso ao nascer (BPN), proxy de ambiente intrauterino hostil, tem sido associado a estas doenças no adulto. Em nosso meio, é escassa a literatura sobre associações de BPN com pressão arterial (PA) e função renal e endócrino-pancreática em adultos. O extenso banco de dados ELSA-Brasil permitiu explorar se: 1) indivíduos pré-diabéticos já poderiam apresentar acometimento renal, detectado de forma precoce por biomarcadores; 2) o BPN associar-se-ia no adulto à PA e marcadores de função renal e endócrino-pancreática mais desfavoráveis que nascidos de peso adequado. Objetivos: Analisar a associação do BPN com PA e marcadores de função renal e endócrino-pancreática em adultos sem DM e sem nefropatia. Os objetivos específicos foram: no Artigo 1, rever a literatura sobre a prevalência de doença renal diabética em indivíduos pré-diabéticos; Artigo 2, analisar a cistatina C sérica (sCys C) como marcador precoce de disfunção renal em indivíduos sem DM do ELSA-São Paulo; Artigo 3, comparar os valores de PA e de marcadores de função renal (taxa de filtração glomerular estimada - TFGe, razão albumina-creatinina - ACR e sCys C) segundo a presença de BPN e analisar sua associação com PA e marcadores de função renal, em indivíduios sem DM e nefropatia; Artigo 4, comparar os valores de marcadores de função de células ß e sensibilidade à insulina (HOMA-ß, HOMA-IR, HOMA-AD, QUICKI, TyG e TG/HDL), segundo a presença de BPN e analisar sua associação com marcadores de função de células ß e sensibilidade à insulina. Métodos: Análises transversais de dados do ELSA-Brasil contemplaram 2 frentes, associação do BPN com PA e função renal e com função pancreática. Foram excluídos indivíduos com >60 anos, IMC <18,5 kg/m², DM, disfunção renal, tireoidiana e hepática. Dados sociodemográficos e de saúde foram coletados por questionários e os clínico-laboratoriais no HU/USP. As variáveis dependentes foram PA, marcadores de função renal e pancreática e a independente o BPN. Variáveis categóricas foram comparadas pelo qui-quadrado e contínuas por teste t de Student ou Wilcoxon. Usou-se regressão linear múltipla para testar associações do peso ao nascer com desfechos e DAG para obter os mínimos ajustes necessários nos modelos. Aplicou-se o escore de propensão para homogeneizar diferenças nos tamanhos amostrais. Resultados: Artigo 1: Verificou-se prevalência de 4,5 a 26,0% de nefropatia diabética no pré-DM. Com base na TFGe, esta taxa variou de 4,5 a 21,3%, na albuminúria de 7,0 a 26,0% e quando combinadas de 12,3 a 17,7%. Artigo 2: Indivíduos com pré-DM tiveram maiores valores de sCys C que os normoglicêmicos [0,67 (0,41-0,95) vs 0,48 (0,31-0,81) mg/L, p<0,001] e menores de TFGe (96,3±17,4 vs 100,6±17,1 mL/min/1,73m², p<0,001). Normoglicêmicos hiperfiltrantes apresentaram valores menores de sCys C que os normofiltrantes (p=0,035). Comparando a TFGe entre os grupos, observou-se queda gradual à medida que pioravam a sCys C e ACR (p-trend=0,06). Artigo 3: O grupo com BPN apresentou níveis mais altos de PA sistólica (p=0,015), diastólica (p=0,014) e de ACR (p=0,031), e menores de TFGe (p=0,015) que o grupo nascido com peso normal. Os prematuros apresentaram níveis médios de PA mais altos que os nascidos com peso normal, mas não houve diferença em marcadores de função renal. À análise de regressão, níveis de PA sistólica e diastólica associaram-se com BPN, mas tal associação não se manteve após incluir prematuridade no modelo, a qual permaneceu associada com PA (p=0,017). Após aplicar escore de propensão, o BPN associou-se com ACR (p = 0.003) mas não com TFGe. Artigo 4: Indivíduos com BPN ou peso normal relataram valores similares de IMC aos 20 anos e o atual foi ligeiramente menor no grupo BPN. Seus dados cardiometabólicos e função endócrino-pancreática foram normais. Em análise de regressão, o HOMA-ß mas não outros índices associou-se ao BPN (p=0,014) independente do sexo, cor, prematuridade e história familiar de DM. Após aplicar o escore de propensão, BPN manteve associação com HOMA-AD e TG/HDL. Conclusão: Nossos achados sugerem que indivíduos com alterações iniciais do metabolismo da glicose já podem apresentar biomarcadores de função renal comprometidos. Atentar-se a eventos precoces da vida como o BPN e prematuridade é relevante, uma vez que associações com PA e biomarcadores de função renal e endócrino-pancreática já podem ser identificadas mesmo em indivíduos saudáveis sem DM e nefropatia. Estudos prospectivos são necessários para avaliar o valor preditivo vislumbrando propor medidas de prevenção.

PK2/PKR1 signaling pathway participates in geniposide protection against diabetic nephropathy in mice / 中国中药杂志

This study aimed to investigate the effects of geniposide(GP) on the expression of prokineticin(PK2) and prokineticin receptor 1(PKR1) in db/db mice with diabetic nephropathy(DN), so as to explore how the PK2 signaling pathway participated in the pathological changes of DN and whether GP exerted the therapeutic effect through this signaling pathway. Male mice were randomly divided into four groups, namely db/m, db/db, db/db+GP, and db/m+GP groups, with five in each group. The mice in the db/db+GP and db/m+GP groups were gavaged with 150 mg·kg~(-1) GP for eight successive weeks. Afterwards, all the mice were sacrificed and the renal tissues were embedded. The morphological changes in glomerulus and renal tubules were observed by Masson and PAS staining. The expression levels of PK2, PKR1, and Wilm's Tumor Protein 1(WT_1) in podocytes were detected by immunohistochemistry, and the protein expression levels of PK2 and PKR1 in mouse kidney by Western blot. The morphological results showed serious glomerular and tubular fibrosis(Masson), high glomerular and tubular injury score(PAS), increased glomerular mesangial matrix, thickened basement membrane, exfoliated brush border of renal tubules, decreased WT_1 in glomerular podocytes, and massive loss of podocytes in the db/db group. After administration with GP, the glomerular and tubular fibrosis was alleviated, accompanied by improved glomerular basement membrane and renal tubule brush edge, and up-regulated WT_1. As revealed by further protein detection, in the db/db group, the expression levels of PK2 and PKR1 and p-Akt/Akt ratio declined, whereas the ratio of Bax/Bcl-2 rose. Ho-wever, PKR2 and p-ERK/ERK ratio did not change significantly. After administration with GP, the PK2 and PKR1 expression was elevated, and p-Akt/Akt ratio was increased. There was no obvious change in PKR2, Bax/Bcl-2 ratio, or p-ERK/ERK ratio. All these have demonstrated that GP improves the renal damage in DN mice, and PK2/PKR1 signaling pathway may be involved in such protection, which has provided reference for clinical treatment of DN with GP.

Clinical comprehensive evaluation of Huangkui Capsules in treatment of chronic kidney diseases / 中国中药杂志

Based on Guidelines for the Management of Clinical Comprehensive Evaluation of Drugs(trial version 2021), this study aims to sort out the clinical evidence of Huangkui Capsules(HC) in the treatment of chronic kidney diseases in aspects of safety, effectiveness, economy, innovation, suitability, accessibility, and characteristics of traditional Chinese medicine( "6+1" dimensions) from real-world data, secondary literature evaluations, questionnaires, and public data, with the methods in evidence-based medicine, epidemiology, pharmacoeconomics, and health technology. Furthermore, with multi-criteria decision analysis(MCDA) model and CSC v2.0, the clinical value of the medicine is comprehensively assessed. All the above are to highlight the advantages and characteristics of HC and lay a basis for scientific decision-making by the medical management department. The dimensions are graded A, B, C, or D. According to the conclusions from phase Ⅳ clinical trial, spontaneous reporting system(SRS), systematic review and Meta-analysis, acute toxicity and long-term toxicity tests, it mainly results in the adverse reactions of nausea, abdominal distension, vomiting, pruritus, rash, and good prognosis in patients. According to the available research, the safety evidence is sufficient and the risk is controllable, so the safety of this medicine is grade B. According to Meta-analysis, HC in combination with conventional drugs in the treatment of chronic kidney disease is superior to conventional drugs alone in reducing urinary protein, serum creatinine concentration, and blood urea nitrogen. In addition, HC combined angiotensin receptor blocker(ARB) or angiotensin converting enzyme inhibitor(ACEI) is outstanding in improving total clinical effective rate, reducing 24 h urinary protein quantity, urinary albumin excretion rate, serum creatinine concentration, triglyceride, and total cholesterol in the treatment of diabetic nephropathy as compared with ARB or ACEI alone. As for chronic nephritis, the application together with ARB or ACEI can raise the total effective rate, reduce 24 h urinary protein content, serum creatinine concentration, and blood urea nitrogen, and delay the progress of the disease. HC boasts high-quality evidence in treating chronic kidney disease, diabetic nephropathy, and chronic nephritis. It has obvious clinical significance in treating chronic kidney disease and thus its efficacy in this aspect is grade B. It has outstanding clinical significance for diabetic nephropathy and chronic nephritis and corresponding and the effectiveness is grade A. As for the pharmacoeconomic value, HC combined with ARB or ACEI is more economical in the treatment of chronic kidney disease than Bailing Capsules combined with ARB or ACEI, with high-quality evidence, and thus the economy of the formula is grade B. HC is a key solution to the high urinary protein in patients with hypotension and chronic kidney disease. The innovation is evidenced by the methods to ensuring drug supply, community-level supply, drug safety, effectiveness, and reasonable price, as wells as the aspects of enterprise philosophy, equipment management, research and development in process and technology, enterprise management and marketing. Thus, the prescription is grade A in innovation. The suitability, as evidenced in drug administration, technical management, drug storage, information service, and medication, is grade B. The course of the medicine is affordable, and it is accessible in a wide range of areas and hospitals. Thus, the accessibility is grade A. HC was developed from an in-hospital preparation, with application in numerous patients and thus large-scale real-world data. As a result, HC is grade B in terms of characteristics of traditional Chinese medicine. After comprehensive evaluation, the clinical value of HC in treating chronic kidney disease is class B, and that for diabetic nephropathy and chronic nephritis is class A. The result is of great reference value for the basic clinical medication management.

Protective effects of Moutan Cortex polysaccharides components on renal injury in diabetic nephropathy rats / 中国中药杂志

This study investigated the protective effects of Moutan Cortex polysaccharides components(MCPC) on the renal tissues of diabetic nephropathy(DN) rats and explored their regulation effect on inflammatory response and oxidative stress. The DN rat model was induced by high-glucose and high-fat diet combined with streptozotocin(STZ), and then the rats were randomly divided into control group, model group, positive group and MCPC high(120 mg·kg~(-1)·d~(-1)), low(60 mg·kg~(-1)·d~(-1)) dose groups. After 12 weeks treatment, blood was taken from the orbit of the rats, and then they were sacrificed before the kidney tissues were collected. The serum and tissues were detected for related biochemical indicators and pathological changes of the kidney. Immunohistochemical methods were used to determine the expression of FN and ColⅣ in the kidney tissue of DN rats. Compared with the model group, blood glucose, serum creatinine, blood urea nitrogen and 24 h urine protein in the MCPC high-dose group were significantly reduced(P<0.01). The results of HE, PAS, Masson staining showed that glomerular basement membrane thickening, Bowman's capsule narrowing and inflammatory cell infiltration in DN rats were improved in the MCPC high-dose group; the activity of T-SOD and GSH-Px in serum significantly increased(P<0.001), and the expression level of FN significantly decreased(P<0.001). The high-dose MCPC treatment could effectively inhibit the abnormal expression of Col Ⅳ(P<0.001) and significantly reduce the levels of AGEs and RAGE in serum(P<0.001), the content of VCAM-1 and IL-1β in serum(P<0.001), and the levels of IL-1β mRNA in kidney tissue(P<0.001), but failed to effectively reduce VCAM-1 mRNA levels in kidney tissues. The high-dose MCPC could significantly improve pathological injury of renal tissue and related renal indicators in DN rats, and achieve renal protection in DN rats mainly by regulating oxidative stress and inflammatory factors.

Normalization of the ratio of nitric oxide and peroxynitrite by promoting eNOS dimer activity is a new direction for diabetic nephropathy treatment / 生理学报

Diabetic nephropathy is a microvascular complication of diabetes. Its etiology involves metabolic disorder-induced endothelial dysfunction. Endothelium-derived nitric oxide (NO) plays an important role in a number of physiological processes, including glomerular filtration and endothelial protection. NO dysregulation is an important pathogenic basis of diabetic nephropathy. Hyperglycemia and dyslipidemia can lead to oxidative stress, chronic inflammation and insulin resistance, thus affecting NO homeostasis regulated by endothelial nitric oxide synthase (eNOS) and a conglomerate of related proteins and factors. The reaction of NO and superoxide (O2.-) to form peroxynitrite (ONOO-) is the most important pathological NO pathway in diabetic nephropathy. ONOO- is a hyper-reactive oxidant and nitrating agent in vivo which can cause the uncoupling of eNOS. The uncoupled eNOS does not produce NO but produces superoxide. Thus, eNOS uncoupling is a critical contributor of NO dysregulation. Understanding the regulatory mechanism of NO and the effects of various pathological conditions on it could reveal the pathophysiology of diabetic nephropathy, potential drug targets and mechanisms of action. We believe that increasing the stability and activity of eNOS dimers, promoting NO synthesis and increasing NO/ONOO- ratio could guide the development of drugs to treat diabetic nephropathy. We will illustrate these actions with some clinically used drugs as examples in the present review.

Maior frequência de neuropatia periférica diabética associada ao hiperparatireoidismo secundário / Higher prevalence of diabetic peripheral neuropathy associated with secondary hyperparathyroidism

INTRODUÇÃO: A neuropatia periférica diabética (ND) é comum na nefropatia diabética (NFD) e não há informações se o hiperparatireoidismo secundário (HPS) aumenta seus sintomas. O objetivo foi determinar ND por sinais em pacientes com HPS. MATERIAL E MÉTODOS: É um estudo caso-controle. O Grupo Controle (GC) é composto por doentes com NFD e valores de paratormônio (PTH)<60pg / ml. O Grupo de Hiperparatireoidismo (GH) engloba pacientes com NFD e PTH≥60pg/ml e critérios bioquímicos de HPS. As variáveis foram, entre outras, a presença de sinais de ND e foram comparados com o teste t de Student e o qui-quadrado. RESULTADOS: Foram 60 participantes em cada grupo, sendo 35 (58,3%) homens no GC vs 33 (55,0%) em GH (p = 0,713). A idade do GC foi de 67±11,0 anos, vs 72±11 anos GH (p=0,009). A taxa de filtração glomerular (TFG) no GC foi 53,82±25,13 vs GH 35,34±18,43ml/min/1,73m2 (p <0,001). O PTH no GC foi de 38,02±15,32 pg/ml, em GH 119,07±84,33 pg/ml (p <0,001). A ND, devido aos sintomas no GC, foi de 28,3% e 36,6% no GH (p=0,330). A neuropatia por sinais no GC foi de 38,3% e no GH 83,3% (p<0,001). O odds ratio de GH para neuropatia presente devido a sinais foi de 8,044 (IC 95% 3,42­8,92). CONCLUSÃO: Constatou-se uma maior presença de sinais de NPD em pacientes com HPS em nosso centro.

BACKGROUND: Diabetic peripheral neuropathy (DN) is common in diabetic nephropathy (DNP), and there is no information if secondary hyperparathyroidism (SHP) increases its symptoms. The purpose was to determine DN by signs in patients with SHP. METHODS: It is a case-control study. Control patients (CG) with DN and parathyroid hormone (PTH) values<60pg/ml. The Hyperparathyroidism group (HG), patients with DNP and PTH≥60pg/ml and HPS biochemical criteria. The variables were, among others, the presence of DN signs, and were compared with Student's t and chi-square. RESULTS: There were 60 participants in each group, 35(58.3%) men in CG vs.33(55.0%) in GH (p=0.713). The age of the CG was 67±11.0 years, vs 72±11years HG (p=0.009). The glomerular filtration rate (GFR) in the CG was 53.82±25.13 vs in HG 35.34±18.43ml/min/1.73m2(p<0.001). The PTH in the CG were 38.02±15.32pg/ml and in GH 119.07±84.33pg/ml(p<0.001). The DN due to symptoms in CG was 28.3% and in GH 36.6%(p=0.330). Neuropathy due to signs in the CG was 38.3% and in GH 83.3% (p<0.001). The HG odds ratio to present neuropathy due to signs was 8.044 (95% CI 3.42­18.92). CONCLUSION: There was a statistical association between HPS and signs of DN in patients with DNP in our canter.

Marein regulates insulin resistance in diabetic nephropathy mice by inducing autophagy / Marein regula la resistencia a la insulina en ratones con nefropatía diabética induciendo la autofagia

SUMMARY: Marein is the main active substance of Coreopsis tinctoria nutt. It not only has anti-oxidation and anti-tumor effects, but also can lower blood lipid, prevent high blood glucose, improve insulin resistance, inhibit gluconeogenesis and promote glycogen synthesis. However, the exact mechanism of its action is still unclear. Here, we explored the effect and mechanism of Marein on insulin resistance. The mice were divided into db/m, db/db, metformin+db/db, and marein+db/db groups. The body weight and kidney weight were recorded. Serum biochemical and renal function tests were measured after 8 weeks of continuous administration. Kidney tissues were subjected to HE staining, PAS staining, and Masson staining. The effect of marein on PI3K/Akt signal and autophagy pathway was detected by Western blot. After 8 weeks of Marein intervention, the body weight and kidney weight of mice did not change significantly, but the fasting blood glucose and blood lipid levels were significantly reduced than db/db group. Marein significantly improved the insulin resistance index, increased serum adiponectin and improved glucose and lipid metabolism disorders of db/db mice. Moreover, marein improved the basement membrane thickness of glomeruli and tubules, improved glomerular sclerosis and tubular fibrosis, as well as renal insufficiency, thereby protecting kidney function and delaying the pathological damage. Furthermore, marein increased the expression of PI3K and the phosphorylation of Akt/Akt (Ser473), and promoted the expression of LC3II/I, Beclin1 and ATG5. Additionally, it promoted the expression of FGFR1 in the kidney of db/db mice, and promoted the increase of serum FGF21 and FGF23. Marein has a protective effect on the kidneys of diabetic mice. It protects diabetic nephropathy by regulating the IRS1/PI3K/Akt signaling pathway to improve insulin resistance. Therefore, marein may be an insulin sensitizer.

RESUMEN: Marein es la principal sustancia activa de Coreopsis tinctoria nutt. No solo tiene efectos antioxidantes y antitumorales, sino que también puede reducir los lípidos en sangre, prevenir la glucemia alta, mejorar la resistencia a la insulina, inhibir la gluconeogénesis y promover la síntesis de glucógeno. Sin embargo, el mecanismo exacto de su acción aún no está claro. Se analizó el efecto y el mecanismo de Marein sobre la resistencia a la insulina. Los ratones se dividieron en grupos db / m, db / db, metformina + db / db y mareína + db / db. Se registró el peso corporal y el peso de los riñones. Se midieron las pruebas de función renal y bioquímica sérica después de 8 semanas de administración continua. Los tejidos renales se sometieron a tinción HE, tinción PAS y tinción Masson. El efecto de la mareína sobre la señal de PI3K / Akt y la vía de autofagia se detectó mediante Western blot. Al término de 8 semanas de tratamiento con mareína, el peso corporal y el peso de los riñones de los ratones no cambiaron significativamente, pero los niveles de glucosa en sangre y lípidos en sangre en ayunas se redujeron significativamente en relación a los del grupo db / db. Marein mejoró significativamente el índice de resistencia a la insulina, aumentó la adiponectina sérica y mejoró los trastornos del metabolismo de la glucosa y los lípidos de los ratones db / db. Además, la mareína mejoró el grosor de la membrana basal de los glomérulos y túbulos, mejoró la esclerosis glomerular y la fibrosis tubular, así como la insuficiencia renal, protegiendo la función renal y retrasando el daño patológico. Además, la mareína aumentó la expresión de PI3K y la fosforilación de Akt / Akt (Ser473), y promovió la expresión de LC3II / I, Beclin1 y ATG5. Además, promovió la expresión de FGFR1 en el riñón de ratones db / db y el aumento de FGF21 y FGF23 en suero. Marein tiene un efecto protector sobre los riñones de ratones diabéticos. Protege la nefropatía diabética regulando la vía de señalización IRS1 / PI3K / Akt para mejorar la resistencia a la insulina. Por tanto, la mareína puede ser un sensibilizador a la insulina.

Assessment of estimated glomerular filtration rate based on cystatin C in diabetic nephropathy / Avaliação da taxa de filtração glomerular estimada com base na cistatina C em nefropatia diabética

Abstract Introduction: GFR is estimated by using creatinine and cystatin C to determine renal dysfunction. Our aim was to evaluate estimated GFR (eGFR) based on cystatin C in type 2 diabetic patients with diabetic nephropathy (DN). Methods: Study group included 52 controls (46% male, age: 54.5±12.4) and 101 diabetic patients (46.5% male, age: 58.2±11). The diabetics were divided into three subgroups according to 24-hour urine albumin: normal to mildly increased (A1) (n=51), moderately increased (A2) (n=25), severely increased (A3) (n=25) albuminuria. Creatinine clearance (CrCl) was determined. Correlations between CrCl and eGFRs estimated according to the CKD-EPI, MDRD, and Cockcroft-Gault (CG) formulas, and ROC curves were evaluated. Data were analyzed using SPSS 22.0. Results: Only CKD-EPI-cys eGFR was significantly lower in the A1 group than the controls (p=0.021). All GFRs were lower in the A3 group than the control (CKD-EPI-cr, MDRD, CKD-EPI-cys, CKD-EPI-cr-cys: p=0.0001, CG and CrCl: p=0.001) and A1 (for all GFRs p=0.0001) groups. CKD-EPI-cr (p=0.004), MDRD (p=0.01), CG (p=0.037), CKD-EPI-cys (p=0.033), and CKD-EPI-cr-cys (p=0.016) eGFRs in the A2 group were significantly different from the A1 group. All eGFRs showed a moderate correlation with CrCl in the A1group (CKD-EPI-cr and CKD-EPI-cr-cys: r=0.49, p=0.0001, MDRD: r=0.44, p=0.001, CG r=0.48, p=0.0001: CKD-EPI-cys r=0.40, p=0.004). The area under the CKD-EPI-cys ROC curve was the highest and found to be 0.847 (95%CI 0.763-0.931, p=0.0001). Conclusions: Our results showed that the CKD-EPI-cys eGFR can be useful in detecting the early stage of DN and more predictive than the others for prediction of DN.

Resumo Introdução: A TFG é estimada usando creatinina e cistatina C para determinar a disfunção renal. Nosso objetivo foi avaliar a TFG estimada (TFGe) com base na cistatina C em pacientes com diabetes do tipo 2 com nefropatia diabética (ND). Métodos: O grupo de estudo incluiu 52 controles (46% homens, idade: 54,5±12,4) e 101 pacientes diabéticos (46,5% homens, idade: 58,2±11). Os diabéticos foram divididos em três subgrupos de acordo com a albumina na urina de 24 horas: albuminúria normal a levemente aumentada (A1) (n=51), moderadamente aumentada (A2) (n=25) e severamente aumentada (A3) (n=25). Foi determinado o clearance de creatinina (Clcr). As correlações entre Clcr e TFGe calculadas de acordo com as fórmulas CKD-EPI, MDRD, e Cockcroft-Gault (CG), e as curvas ROC foram avaliadas. Os dados foram analisados usando o SPSS 22.0. Resultados: Somente a TFGe CKD-EPI-cis foi significativamente menor no grupo A1 do que nos controles (p=0,021). Todas as TFGs foram mais baixas no grupo A3 do que no grupo controle (CKD-EPI-cr, MDRD, CKD-EPI-cis, CKD-EPI-cr-cis: p=0,0001, CG e Clcr: p=0,001) e no grupo A1 (para todas as TFGs p=0,0001). As TFGes CKD-EPI-cr (p=0,004), MDRD (p=0,01), CG (p=0,037), CKD-EPI-cis (p=0,033), e CKD-EPI-cr-cis (p=0,016) no grupo A2 foram significativamente diferentes do grupo A1. Todas as TFGes mostraram uma correlação moderada com Clcr no grupo A1 (CKD-EPI-cr e CKD-EPI-cr-cis: r=0,49, p=0,0001, MDRD: r=0,44, p=0,001, CG r=0,48, p=0,0001: CKD-EPI-cis r=0,40, p=0,004). A área sob a curva ROC CKD-EPI-cis foi a mais alta e foi considerada 0,847 (95%IC 0,763-0,931, p=0,0001). Conclusões: Nossos resultados mostraram que a TFGe CKD-EPI-cis pode ser útil na detecção do estágio inicial de ND e com maior valor de predição do que as outras para a predição da ND.

Effect of Huangkui on mouse podocytes under high glucose environment / Efecto de Huangkui en podocitos de ratón en un entorno de glucosa alta

To explore a new underlying molecular mechanism of Huangkui Extract Powder (HKEP) in the alleviation of diabetic nephropathy (DN). Murine immortalized podocytes were divided into (i) normal glucose (NG, 5.6 mM), (ii) NG + HKEP (0.45 g/L), (iii) HG, and (iv) HG + HKEP (0.45 g/L) groups. MTT assay and flow cytometry were used to detect the podocyte proliferation, apoptosis and cell cycle. Cell viability was inhibited, and apoptosis increased in(iii) HG group compared with (i) NG group (p<0.05). mRNA and protein expression of nephrin and podocin significantly decreased in (iii) HG group compared with (i) NG group (p<0.05). When compared with (iii) HG group, (iv) HG + HKEP group had higher cell viability, lower apoptotic rate and higher mRNA and protein expression of nephrin and podocin (p<0.05). HKEP can attenuate HG-induced podocyte damage, which may be one of the mechanisms of HKEP for attenuating DN.

Explorar un nuevo mecanismo molecular subyacente del extracto del polvo de Huangkui (HKEP) en el alivio de la nefropatía diabética (ND). Los podocitos murinos inmortalizados se dividieron en (i) grupos de glucosa normal (NG, 5,6 mM), (ii) NG + HKEP (0,45 g/L), (iii) HG y (iv) HG + HKEP (0,45 g/L). Se utilizaron el ensayo MTT y la citometría de flujo para detectar la proliferación de podocitos, la apoptosis y el ciclo celular. La viabilidad celular se inhibió y la apoptosis aumentó en el grupo (iii) HG en comparación con el grupo (i) NG (p<0,05). La expresión de ARNm y proteínas de nefrina y podocina disminuyó significativamente en el grupo (iii) HG en comparación con el grupo (i) NG (p<0,05). En comparación con el grupo (iii) HG, el grupo (iv) HG + HKEP tuvo una mayor viabilidad celular, una tasa de apoptosis más baja y una expresión de ARNm y proteínas más altas de nefrina y podocina (p<0,05). HKEP puede atenuar el daño de los podocitos inducido por HG, que puede ser uno de los mecanismos de HKEP para atenuar la DN.

Investigation of the protective and therapeutic effects of ß-aminoisobutyric acid (BAIBA) and Thymoquinone in the diabetic nephropathy / Investigación de los efectos protectores y terapéuticos del ácido ß-aminoisobutírico (BAIBA) y la timoquinona en la nefropatía diabética

In this study, against streptozotocin (STZ) induced diapetic nephropathy (DN); it is aimed to investigate the use of thymoquinone (TQ) and ß-aminoisobutyric acid (BAIBA) and to compare the effects of these agents. With random selection of 35 male rats, five groups (seven rats in each group) were constituted as follows: Control, STZ, STZ + TQ, STZ + BAIBA, STZ + TQ + BAIBA. In the STZ group; body weight, glutathione (GSH) and insulin levels decreased, relative kidney weight, malondialdehyde (MDA), glucose, blood urea nitrogen (BUN) and creatinine (Cr) levels were increased. Also, in kidney tissue; histopathological changes (such as thickening of the capsular, glomerular and tubular basement membranes, increased mesangial matrix amount, increased cytoplasmic vacuolization in some of the tubular epithelial cells, increased tumor necrosis factor-alpha (TNF-α) expression, and inflammatory cell infiltrations in interstitial tissue) were detected. It was observed that these changes occurring after diabetes mellitus (DM) reversed significantly in TQ, BAIBA and TQ + BAIBA groups.

En este estudio, contra la nefropatía diapética (ND) inducida por estreptozotocina (STZ); tiene como objetivo investigar el uso de timoquinona (TQ) y ácido ß-aminoisobutírico (BAIBA) y comparar los efectos de estos agentes. Con la selección aleatoria de 35 ratas macho, se constituyeron cinco grupos (siete ratas en cada grupo) como sigue: Control, STZ, STZ + TQ, STZ + BAIBA, STZ + TQ + BAIBA. En el grupo STZ; el peso corporal, los niveles de glutatión (GSH) y de insulina disminuyeron, el peso relativo de los riñones, el malondialdehído (MDA), la glucosa, el nitrógeno ureico en sangre (BUN) y los niveles de creatinina (Cr) aumentaron. Además, en tejido renal; se detectaron cambios histopatológicos (como engrosamiento de las membranas basales capsular, glomerular y tubular, aumento de la cantidad de matriz mesangial, aumento de la vacuolización citoplasmática en algunas de las células epiteliales tubulares, aumento de la expresión del factor de necrosis tumoral alfa (TNF-α) e infiltraciones de células inflamatorias en tejido intersticial). Se observó que estos cambios que ocurren después de la diabetes mellitus (DM) se revirtieron significativamente en los grupos TQ, BAIBA y TQ + BAIBA.

Influence of long non-coding RNA Gm15645 on the podocyte injury in diabetic nephropathy / 中华医学遗传学杂志

OBJECTIVE@#To explore the influence of long non-coding (lnc) RNA Gm15645 on the podocyte injury in mice with diabetic nephropathy.@*METHODS@#Male db/db mice (with Type 2 diabetes) with a genetic background of C57BLKs/J and db/m mice (healthy) born in littermates were randomly divided into three groups. db/db group was injected with lncRNAGm15645 shRNA lentivirus with a podocyte-specific marker NPHS2; db/db blank group was injected with saline, and db/db control group was injected withnon-sense lentivirus. The results of PAS staining, pathological changes of renal tissue, relative expression of GSK-3beta, and podocin expression were compared.@*RESULTS@#lncRNAGm15 645 was overexpressed and podocin was down-regulated in the lentivirus overexpressed group. Mesangial cell proliferation, mesangial matrix hyperplasia, thickened basement membrane, widely fused foot process, and podocyte injury were observed by PAS staining. The expression of Gm15645 in the db/db group was significantly lower than that of the db/db blank group and db/db control group (P< 0.05), while the expression of podocin was higher (P< 0.05). Gm15645 was co-stained with podocin in renal tissue, and the target gene was GSK-3beta.@*CONCLUSION@#lncRNAGm15645 may provide an early biomarker for the occurrence of podocyte injury in diabetic nephropathy. The mechanism may be related to the feedback regulation of GSK-3beta gene.

Inibição combinada do receptor AT1 e produtos finais de glicação avançada na nefropatia diabética / Combined inhibition of AT1 receptor and advanced glycation end products in diabetic nephropathy

O diabetes mellitus (DM) é um grande problema de saúde pública, que afeta cerca de 463 milhões de pessoas no mundo e pode alcançar 700 milhões de pessoas até 2045. A nefropatia é uma das complicações microvasculares do diabetes e a maior causa de insuficiência renal. Uma vez que a ativação do receptor AT1 pela angiotensina II causa vasoconstrição da artéria aferente, e que o acúmulo de produtos finais de glicação avançada (AGEs) causam glicotoxicidade intracelular de células mesangiais, podocitárias e tubulares, foi avaliado se o tratamento combinado de olmersatana (OLM) e piridoxamina (PYR) é capaz de melhorar a nefropatia diabética quando comparado aos tratamentos isolados. Para isto, o diabetes mellitus experimental foi induzido em 50 camundongos C57BL/6 pela administração de estreptozotocina (50 mg/kg/dia via intraperitoneal por 5 dias). Os animais foram divididos em cinco grupos: controle, diabéticos, diabéticos tratados com OLM (20 mg/Kg/dia), diabéticos tratados com PYR (400 mg/Kg/dia) e diabéticos tratados com OLM e PYR (20 mg/Kg/dia e 400 mg/Kg/dia, respectivamente) Os tratamento durou 16 semanas. Como resultado, os camundongos que receberam STZ desenvolveram hiperglicemia e doença renal, diminuição de peso, poliúria, polidipsia, polifagia, e albuminúria, além de aumento de frutosamina, ferro, uréia e fosfatase alcalina na urina, diminuição da atividade de catalase no rim e aumento de excreção dos AGEs na urina. Histologicamente, houve aumento de área glomerular e do espaço de BowmanO tratamento com OLM atenuou a albuminúria e atenuou o declínio da função renal, uma vez que o OLM melhorou a polidipsia, polifagia, poliúria, marcadores bioquímicos de disfunção renal e parâmetros histológicos, apesar de não apresentar ação sobre a fosfatase alcalina, uréia, ferro, frutosamina e ácido úrico. O tratamento isolado com PYR melhorou a maioria dos parâmetros alterados pela doença, entre eles a ingestão alimentar, ingestão hídrica, volume urinário, creatinina sérica, excreção de albumina na urina, ACR, AGEs no tecido renal e urina, ureia na urina e todos os parâmetros morfológicos analisados, contudo sem efeito sobre a fosfatase alcalina, ferro, frutosamina e ácido úrico. O tratamento combinado (PYR e OLM) melhorou a polifagia, polidipsia e poliúria comparado aos animais diabéticos sem tratamento, entretanto não houve diferença comparado a cada tratamento isolado. Os parâmetros histológicos e bioquímicos (creatinina no soro, albumina na urina, ACR e uréia) também apresentaram melhorara em relação aos animais diabéticos sem tratamento, mas não em relação aos animais em tratamento isolado. Tanto PYR quanto OLM não influenciaram a excreção de AGEs na urina. Esses dados nos levam a concluir que o tratamento combinado não ofereceu efeitos benéficos adicionais quando comparado aos tratamentos isolados, e que o tratamento que mais ofereceu benefícios foi o tratamento isolado com PYR nos parâmetros metabólicos, morfológicos e de função renal. (AU)