Construction of a TF-miRNA-mRNA Regulatory Network for Diabetic Nephropathy

Background: This study aims to elucidate the microRNA (miRNA)-messenger RNA (mRNA)-transcription factors (TFs) network relevant to diabetic nephropathy (DN). Methods: To investigate the molecular mechanisms underlying DN, we conducted an extensive analysis using a Gene Expression Omnibus (GEO) database, specifically GSE51784, GSE30528, GSE30529 and GSE1009. RNA samples from 66 subjects were analysed to identify differentially expressed mRNAs (DEGs) and microRNAs (DEMs) between individuals with DN and healthy controls. The data underwent preprocessing, followed by Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Set Enrichment Analysis (GSEA) to unveil enriched pathways. Additionally, we constructed protein-protein interaction networks and subnetworks of modules to identify key molecular players. Results: A total of 163 DEMs and 188 DEGs were identified among the four datasets. Furthermore, we identified 37 hub genes with high connectivity and four TFs, namely E1A Binding Protein P300 (EP300), SP100 Nuclear Antigen (SP100), Nuclear Receptor Subfamily 6 Group A Member 1 (NR6A1) and Jun Dimerization Protein 2 (JDP2), which may play crucial roles in the molecular pathogenesis of DN. Additionally, we constructed a co-regulatory network involving miRNAs, mRNAs and TFs, revealing potential involvement of pathways such as the Mitogen-Activated Protein Kinase (MAPK) signalling pathway, phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signalling pathway and metabolic pathways in the pathogenesis of DN. Finally, using a docking model, we established drug-gene interactions involving key genes in the network, providing potential insights into therapeutic options. Conclusions: This study explores a gene regulation network of miRNA-mRNA-TFs, identifying potential molecular targets in the aetiology of DN. It also suggests potential targets for genetic counselling and prenatal diagnosis for DN (AU)

MTF1 has the potential as a diagnostic and prognostic marker for gastric cancer and is associated with good prognosis

Purpose Metal Regulatory Transcription Factor 1 (MTF1) can be an essential transcription factor for heavy metal response in cells and can also reduce oxidative and hypoxic stresses in cells. However, the current research on MTF1 in gastric cancer is lacking. Methods Bioinformatics techniques were used to perform expression analysis, prognostic analysis, enrichment analysis, tumor microenvironment correlation analysis, immunotherapy Immune cell Proportion Score (IPS) correlation and drug sensitivity correlation analysis of MTF1 in gastric cancer. And qRT-PCR was used to verify MTF1 expression in gastric cancer cells and tissues. Results MTF1 showed low expression in gastric cancer cells and tissues, and low expression in T3 stage compared with T1 stage. KM prognostic analysis showed that high expression of MTF1 was significantly associated with longer overall survival (OS), FP (first progression) and PPS (post-progression survival) in gastric cancer patients. Cox regression analysis showed that MTF1 was an independent prognostic factor and a protective factor in gastric cancer patients. MTF1 is involved in pathways in cancer, and the high expression of MTF1 is negatively correlated with the half maximal inhibitory concentration (IC50) of common chemotherapeutic drugs. Conclusion MTF1 is relatively lowly expressed in gastric cancer. MTF1 is also an independent prognostic factor for gastric cancer patients and is associated with good prognosis. It has the potential to be a diagnostic and prognostic marker for gastric cancer (AU)

Identification of Hub Genes Associated with Progression and Prognosis of Bladder Cancer by Integrated Bioinformatics Analysis

Background: Bladder cancer (BLCA) is an extremely common carcinoma of the urinary system that has a high incidence of relapse. Although intensive studies have investigated its pathology in the past decades, there are significant knowledge gaps regarding the characterization of the molecular processes underlying the progression of disease and consequently its prognosis. The purpose of current research was to identify significant genes that could serve as prognostic and progression biomarkers. Methods: Gene expression profiles were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database. Differential gene expression analysis (DGEA) and weighted gene co-expression network analysis (WGCNA) were conducted to recognize differential co-expression genes. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to explore gene function. Moreover, protein-protein interactions (PPI) network, overall survival (OS) and disease-free survival (DFS) were used to identify survival-related hub genes. Additionally, associations between these gene’s expression and clinical parameters were determined. Finally, the Human Protein Atlas (HPA) database and qRT-PCR were used to validate gene’s expression. Results: About 124 differential co-expression genes were identified. These genes were mainly enriched in muscle system process and muscle contraction (biological process, BP), contractile fiber, myofibril, sarcomere, focal adhesion and cell-substrate junction (cellular component, CC) and actin binding (molecular function, MF) in GO enrichment analysis, while enriched in vascular smooth muscle contraction, focal adhesion, cardiac muscle contraction, hypertrophic cardiomyopathy, dilated cardiomyopathy and regulation of actin cytoskeleton in KEGG analysis (AU)

La epigenética. Sus mecanismos y significado en la regulación génica / Epigenetics. Mechanism and significance in gene regulation

La epigenética trata del estudio de las modificaciones estructurales en las regiones del genoma, por metilación del ADN o de las histonas cromosómicas, u otros mecanismos que afectan a la expresión de los genes sin alterar la composición de bases del ADN. La diferenciación celular, que se establece en el desarrollo embrionario a partir del estado de blastocisto, es la consecuencia de una reprogramación celular diferencial, basada en modificaciones epigenéticas programadas, que establecen diferencias en el epigenoma de cada célula y tejido. Los genomas maternos y paternos de los gametos tienen diferencias de impresión genómicas debido a la metilación del ADN u otras modificaciones epigenéticas establecidas en las células germinales durante la gametogénesis. También pueden producirse modificaciones epigenéticas no programadas bajo la influencia de factores ambientales no controlados, que pueden dar lugar a alteraciones de la salud. Sin embargo, todas las modificaciones epigenéticas se borran tras la fecundación y las que afectasen al tejido germinal del embrión o el feto durante su desarrollo no se heredan más allá de la segunda generación -la F2. La epigenética no debe considerarse un nuevo tipo de herencia con consecuencias transgeneracionales, sino como un conjunto de mecanismos relacionados con la regulación genética

Epigenetics deals with the study of structural modifications in the regions of the genome, by methylation of DNA or chromosomal histones, or other mechanisms that affect the expression of genes without altering the base composition of DNA. Cell differentiation, which is established in embryonic development from blastocyst status, is the consequence of differential cell reprogramming, based on programmed epigenetic modifications, which establish differences in the epigenome of cells and tissues. Maternal and paternal genomes of the gametes have genomic imprinting differences due to DNA methylation or other epigenetic modifications established in the germinal cells during gametogenesis. Maternal and paternal parental genomes present differences of genomic imprinting, which are established by DNA methylation or other epìgenetic modifications during embryonic development, in the primordial germ cells. Unscheduled epigenetic modifications may also occur under the influence of uncontrolled environmental factors, which can lead to health disturbances. However, all epigenetic modifications are erased after fertilization and those affecting the germ line of the embryo or fetus during their development are not inherited be-yond the second generation, -F2-. Epigenetics should not be considered a new type of inheritance with transgenerational consequences, but as a set of mechanisms related to genetic regulation

Evaluation of miR-181b and miR-126-5p expression levels in T2DM patients compared to healthy individuals: Relationship with NF-KappaB gene expression / Evaluación de los niveles de expresión de miR-181b y miR-126-5p en pacientes con DMT2 comparados con personas sanas: relación con la expresión del gen NF- kappaB

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a progressive metabolic disorder whose prevalence is rising very fast across the world. Diagnosis of this disease in early stages (pre-diabetic stage) plays an important role in reducing mortality associated with this disorder. miRNAs, as key players in the pathogenesis of T2DM, have been investigated in several studies. Furthermore, their expression profile changes in the early stages of diabetes mellitus in body fluids such as serum, peripheral blood, and peripheral blood mononuclear cell (PBMC) have been studied. Due to their high stability and the presence of non-invasive sensitive methods for their measurement, such as real-time PCR, they can be used for early diagnosis of T2DM as a biomarker. In this experimental study, the expression levels of miR-181b, miR-126-5p, and NF-KappaB were measured in patients with T2DM, pre-diabetic subjects, and healthy controls in a Yazd population. MATERIAL AND METHOD: Ninety asymptomatic subjects including 30 T2DM, 30 pre-diabetic, and 30 healthy subjects (diagnosis based on WHO criteria) were included in this study. Real-time PCR was used to measure the expression levels of miR-181b and miR-126-5p. Moreover, the NF-KappaB expression level was also measured to determine its relationship with these two microRNAs. RESULT: In this study, the expression level of miR-181b and miR-126-p decreased gradually in pre-diabetic as well as T2DM subjects compared to healthy controls. Furthermore, our study showed a significant negative correlation between these two miRNAs and NF-KappaB for the first time. CONCLUSION: These results introduce these anti-inflammatory miRNAs as powerful tools for early diagnosis of T2DM

ANTECEDENTES: La diabetes mellitus tipo 2 (DMT2) es un trastorno metabólico progresivo cuya prevalencia aumenta muy rápidamente en todo el mundo. El diagnóstico de esta enfermedad en estadios iniciales (fase prediabética) tiene un papel importante para reducir la mortalidad asociada con este trastorno. Los miARN, como elementos clave en la patogenia de la DMT2, se han investigado en varios estudios. Además, se han estudiado los cambios de su perfil de expresión en los estadios iniciales de la diabetes mellitus en líquidos corporales como el suero, la sangre periférica y las células mononucleares de sangre periférica (CMSP). Gracias a su elevada estabilidad y a la existencia de métodos sensibles no invasivos para medirlos, como la RCP en tiempo real, pueden utilizarse como biomarcadores para el diagnóstico precoz de la DMT2. En este estudio experimental se determinaron los niveles de expresión de miR-181b, miR-126-5p y NF-kappaB en pacientes con DMT2, sujetos prediabéticos y controles sanos de una población de la ciudad de Yazd. MATERIAL Y MÉTODO: Se incluyeron en este estudio a 90 sujetos asintomáticos, incluidos 30 con DMT2, 30 prediabéticos y 30 sanos (el diagnóstico se basó en los criterios de la OMS). Se utilizó RCP en tiempo real para determinar los niveles de expresión de miR-181b y miR-126-5p. Se midió también el nivel de expresión de NF-kappaB para determinar su relación con estos 2 micro-ARN. RESULTADOS: En este estudio, el nivel de expresión de miR-181b y miR-126-5p descendió gradualmente en los sujetos prediabéticos y con DMT2 comparados con los controles sanos. Además, nuestro estudio mostró por primera vez una correlación negativa importante entre estos 2 miARN y NF-kappaB. CONCLUSIÓN: Estos resultados sugieren que estos miARN antiinflamatorios son herramientas potentes para el diagnóstico precoz de la DMT2

The expression of DNMTs is dramatically decreased in peripheral blood mononuclear cells of male patients with juvenile idiopathic arthritis

INTRODUCTION: Juvenile idiopathic arthritis (JIA) is an autoimmune rheumatic disease, which affects primarily the joints in children under 16 years old. The etiology of JIA is yet unknown but research has shown that JIA is a multifactorial disease implicating several genes and environmental factors. Environmental factors affect immune cells via epigenetic mechanisms. One of the most important epigenetic mechanisms is DNA methylation catalyzed by DNA methyltransferases (DNMTs) and usually associated with gene silencing. In this study, we analyzed the expression of three DNA methyltransferases namely DNMT1, DNMT3a and DNMT3b in peripheral blood mononuclear cells (PBMCs) of patients with JIA and compared it with the expression of these genes in healthy young individuals. MATERIALS AND METHODS: Peripheral blood mononuclear cells of 28 JIA patients and 28 healthy controls were isolated. Total RNA was extracted, cDNA was synthesized and the transcript levels of DNMTs were analyzed by quantitative PCR. RESULTS: Analysis of DNMT1, DNMT3a and DNMT3b relative gene expression in PBMCs of JIA patients and control individuals shows that the expression of DNMT1 and DNMT3a is reduced significantly by 7 folds and 5.5 folds, respectively, in JIA patients compared to healthy controls. Furthermore, the expression of all three DNMTs were significantly and drastically reduced in young affected males compared to healthy males. CONCLUSION: This study shows that the expression of DNMTs is reduced in JIA patients and this reduction is severe in male JIA patients

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Regulación por micro RNA de la respuesta angiogénica en la retina diabética / Micro-RNA regulation of the angiogenic response in the diabetic retina

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Impacto funcional de polimorfismos del gen de la esclerostina sobre la metilación de ADN y la expresión génica / Functional impact of sclerostin gene polymorphisms on DNA methylation and gene expresión

INTRODUCCIÓN: Varios estudios de barrido genómico (GWAS) y otros focalizados en el gen de la esclerostina (SOST) han encontrado que algunos polimorfismos de SOST se asocian con la masa ósea y el riesgo de fracturas. El objetivo de este estudio fue analizar la relevancia funcional de ciertos polimorfismos de la región promotora de SOST, en relación con la expresión y la metilación de dicho gen. MATERIAL Y MÉTODO: Para ello, se determinaron los alelos de los polimorfismos rs851054, rs851056, rs10534024, rs1234612 y se analizó la metilación de ADN de 33 muestras de suero y de hueso, procedentes de pacientes intervenidos para colocar una prótesis de cadera, mediante pirosecuenciación tras conversión con bisulfito. Además, en el hueso se estudió la expresión de SOST. Por último, se clonaron diferentes alelos del promotor de SOST en vectores reporteros dobles con el gen de la luciferasa bajo dicho promotor y el gen de la fosfatasa alcalina bajo un promotor constitutivo. RESULTADOS: El análisis de metilación de la región promotora de SOST en ADN libre en suero y en ADN de hueso no reveló diferencias estadísticamente significativas en relación con los alelos de los polimorfismos analizados (p > 0,05). Sin embargo, las transfecciones con los vectores reporteros mostraron una elevada actividad transcripcional, independientemente del vector utilizado. CONCLUSIÓN: No hemos encontrado una asociación clara entre los distintos alelos y la metilación de ADN de la región promotora del gen SOST. Son necesarios más estudios para determinar los efectos funcionales de los polimorfismos sobre la metilación y expresión del gen de SOST y los efectos sobre la masa ósea

INTRODUCTION: Several genome‐wide association studies (GWAS) and others which focused on the sclerostin gene (SOST)have found that some SOST polymorphisms are associated with bone mass and risk of fractures. This study analyzes thefunctional relevance of certain polymorphisms of the SOST promoter region, and their relationship with the expressionand methylation of this gene. MATERIAL AND METHODS: Alleles of the rs851054, rs851056, rs10534024, rs1234612 polymorphisms and DNA methylationwere analyzed by pyrosequencing in serum and bone samples of 33 patients undergoing hip replacement. In addition,SOST expression was studied in bone samples. Also, different alleles of the SOST promoter were cloned into double reportervectors with the luciferase gene under this promoter and the alkaline phosphatase gene under a constitutive promoter. RESULTS: Methylation analysis of the SOST promoter region in serum free DNA and bone DNA revealed no statisticallysignificant differences across the alleles of the analyzed polymorphisms (p > 0.05). However, transfections with reportervectors showed high transcriptional activity, regardless of the vector used. CONCLUSION: We have not found a clear association between the different alleles and the DNA methylation of the SOSTpromoter region. Further studies are needed to determine the polymorphisms' functional effects on the methylationand expression of the SOST gene and the consequences on bone mass

Effect of ginger extract on expression of GATA3, T-bet and ROR-yt in peripheral blood mononuclear cells of patients with Allergic Asthma

Introduction and objectives: Allergic asthma is a chronic inflammatory disorder of the airways. Th1, Th2 and Th17 cells are the main cells involved in the pathophysiology of asthma. The function of these cells is affected by T-bet, GATA3 and RORgammat transcription factors (respectively). Therefore, the aim of this study was to evaluate the effect of ginger (officinal Roscoe) extract on the expression of T-bet, GATA-3 and ROR-gamma in peripheral blood mononuclear cells (PBMC) of asthmatic patients, in comparison with healthy volunteers as controls. Materials and methods: In this case-control study, a total of 50 individuals including 25 patients with severe, moderate and mild allergic asthma and 25 unrelated healthy controls were involved. The PBMCs were isolated and divided into four groups: negative control, two positive controls (Budesonide and PHA) and ginger-extract treated group. After cell treatment and incubation for 48h, PBMCs were isolated and cDNA was synthesized. Gene expressions of T-bet, GATA3 and ROR-γt were evaluated by Real-time PCR. Results: According to the results of this study, hydroalcoholic extract of ginger could reduce the expression of GATA-3, ROR-gammat, and T-bet in PBMCs of asthmatic patients in comparison with untreated PBMCs (P values = 0.001, 0.001, and 0.002, respectively). It was also shown that the ginger extract could affect T-bet/GATA-3, T-bet/ROR-gamma, and ROR-gammat/GATA-3 expression ratios. Conclusions: This study showed that the use of ginger extract could control asthma and decrease the severity of this disease by affecting the main cells involving the symptoms of asthma in the airways

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Impact of green tea epigallocatechin-3-gallate on HIF1-α and mTORC2 expression in obese women: anti-cancer and anti-obesity effects? / Impacto del té verde epigalocatequina-3-galato en la expresión de HF1-a y mTORC2 en mujeres obesas: ¿efectos anticancerígenos y antiobesidad?

Introduction: epigallocatechin-3-gallate (EGCG) is the most abundant catechin contained in green tea (Camellia sinensis) and has been associated with anti-obesity and anti-cancer effects, but the exact molecular mechanisms remain elusive. In this context, this study was designed to improve the understanding of the EGCG anti-obesity and anti-cancer action. Objectives: this study was designed to examine the effects of EGCG on the expression of genes involved in obesity and cancer pathways in the peripheral blood mononuclear cells of obese women. Material and methods: this longitudinal interventional study enrolled eleven women with severe obesity that were submitted to eight weeks of green tea (decaffeinated green tea capsules with 450.7 mg of EGCG, two capsules/day) supplementation (intervention group) and ten eutrophic women as a control group. Weight (kg), body mass index (BMI, kg/m2), fat mass (kg) and gene expression (qPCR method) were assessed before and after supplementation. HIF1-alpha (HIF1-α), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) and rapamycin-insensitive companion of mTOR (RICTOR) were selected as potential targets. Results: after supplementation, body weight (114.9 ± 14.3 versus 115 ± 13.8 kg), body mass index (44.1 ± 3.7 versus 44.1 ± 3.9 kg/m2) and fat mass (47.6 ± 3.3 versus 47.3 ± 3.4 kg) did not change. EGCG upregulated the RICTOR and HIF1-α expression, however, did not modify PI3K expression. Conclusion: this study demonstrated that EGCG has a potential role to obesity and cancer related to obesity control and can be used not only for the purpose of weight loss, but also for the improvement of obesity-related comorbidities

Introducción: la obesidad se asocia con altos niveles de estrés oxidativo (EO) e inflamación. Existe mucha evidencia de que algunos polifenoles, como el té verde, tienen un impacto positivo en el estado del sistema operativo y consecutivamente en la inflamación. Objetivos: los propósitos de este estudio fueron: a) acceso a biomarcadores de EO en mujeres obesas y de peso normal; y b) evaluar si la suplementación con té verde tiene impacto en los biomarcadores de citoquinas inflamatorias y de EO de mujeres obesas. Métodos: evaluamos mujeres obesas (índice de masa corporal - IMC ≥ 40 kg/m²) y peso normal (IMC entre 18,5 y 24,9 kg/m²). Se utilizaron muestras de sangre para acceder al malondialdehído (MDA), la capacidad antioxidante equivalente de Trolox (TEAC) y las citoquinas inflamatorias. Elegimos al azar pacientes obesos (18 individuos) y luego les dimos suplementos de té verde durante 8 semanas. El análisis estadístico incluyó las pruebas de Shapiro-Wilk, Wilcoxon, t pareadas e independientes, p < 0,05 se consideraron significativas. Resultados: se reclutaron 42 mujeres obesas (IMC: 48,2 ± 9,3 kg/m2) y 21 de peso normal (IMC: 22,5 ± 2 kg/m2) con una edad promedio de 36,2 ± 9,1 años. Los niveles séricos de MDA fueron más altos en las personas obesas (2,52 ± 0,31 μmol/L) que en las mujeres eutróficas (2,13 ± 0,26 μmol/L; p = 0.000). Por otro lado, se observaron valores de TEAC más bajos en obesos (0,75 ± 0,06 mM) que en el grupo eutrófico (0,78 ± 0,04 mM; p = 0,009). Después de la intervención del té verde, la MDA disminuyó 4,7% y el TEAC aumentó 10%. Los niveles séricos de interleucina-6 (IL-6) disminuyeron 12,7% después del tratamiento (p = 0,03). Conclusiones: el grupo obeso tenía menor capacidad antioxidante que el eutrófico. La suplementación con té verde mejoró TEAC y MDA y redujo los niveles séricos de IL-6 en mujeres obesas

Stability of asthma control implies no changes in microRNAs expression

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Alpha-syntrophin dependent expression of tubulin alpha 8 protein in hepatocytes

The scaffold protein alpha-syntrophin (SNTA) is a component of the dystrophin glycoprotein complex and has been comprehensively studied in skeletal muscle and adipocytes. SNTA is further expressed in the liver where its biological role remains unclear. Unpublished data from our group suggested that SNTA deficiency is associated with altered tubulin alpha 8 (TUBA8) levels in fat. TUBA8 is highly expressed in different cell lines including hepatoma cells, and here we analyzed whether SNTA has a role herein. In Hepa1-6 cells, TUBA8 protein levels were increased upon SNTA knock down and were reduced upon overexpression of SNTA. This regulation was not identified when analyzing mRNA expression. In the liver of SNTA-deficient mice, TUBA8 protein was higher compared to the respective wild-type controls while RNA expression was even suppressed. Using the HaloTag platform, TUBA8 was found to form a complex with SNTA in Hepa1-6 cells. In the hepatic stellate cell line LX-2, the lack or overexpression of SNTA did, however, not change TUBA8 protein expression. SNTA and TUBA8 are described to regulate cell proliferation. Yet, knock down of SNTA did neither affect proliferation nor viability of Hepa1-6 cells. The present study shows that SNTA protein levels are inversely related to TUBA8 protein expression in the hepatocyte cell line Hepa1-6

Effects of exosomes from LPS-activated macrophages on adipocyte gene expression, differentiation, and insulin-dependent glucose uptake

Obesity is usually associated with low-grade inflammation, which determines the appearance of comorbidities like atherosclerosis and insulin resistance. Infiltrated macrophages in adipose tissue are partly responsible of this inflammatory condition. Numerous studies point to the existence of close intercommunication between macrophages and adipocytes and pay particular attention to the proinflammatory cytokines released by both cell types. However, it has been recently described that in both, circulation and tissue level, there are extracellular vesicles (including microvesicles and exosomes) containing miRNAs, mRNAs, and proteins that can influence the inflammatory response. The objective of the present research is to investigate the effect of exosomes released by lipopolysaccharide (LPS)-activated macrophages on gene expression and cell metabolism of adipocytes, focusing on the differential exosomal miRNA pattern between LPS- and non-activated macrophages. The results show that the exosomes secreted by the macrophages do not influence the preadipocyte-to-adipocyte differentiation process, fat storage, and insulin-mediated glucose uptake in adipocytes. However, exosomes induce changes in adipocyte gene expression depending on their origin (LPS- or non-activated macrophages), including genes such as CXCL5, SOD, TNFAIP3, C3, and CD34. Some of the pathways or metabolic processes upregulated by exosomes from LPS-activated macrophages are related to inflammation (complement activation, regulation of reactive oxygen species, migration and activation of leukocyte, and monocyte chemotaxis), carbohydrate catabolism, and cell activation. miR-530, chr9_22532, and chr16_34840 are more abundant in exosomes from LPS-activated macrophages, whereas miR-127, miR-143, and miR-486 are more abundant in those secreted by non-activated macrophages

Evidence for the link between defective autophagy and inflammation in peripheral blood mononuclear cells of type 2 diabetic patients

Autophagy was shown to modulate inflammation in immune cells. This study was designed to evaluate the association between autophagy and inflammation in peripheral blood mononuclear cells (PBMCs) of type 2 diabetic (T2D) and non-diabetic (ND) subjects. The autophagy markers were measured by real-time PCR and western blot. The gene expression of pro- and anti-inflammatory cytokines was assessed by real-time PCR. Reduced transcription of BECN1 and LAMP2 and unchanged expression of MAP1LC3B and ATG5 were observed in PBMCs of T2D patients. Decreased LC3B-II and increased p62/SQSTM1 levels were found in PBMCs of diabetic patients. The p-mTOR level was higher in PBMCs of diabetic patients. An increase in both IL-1Beta and TNF-alfa gene expression, along with a decrease in the expression of IL-10, was observed in PBMCs of T2D patients. TNF-α mRNA expression was inversely correlated with the mRNA expression of BECN1 and LAMP2. TNF-alfa and IL-1Beta expression were negatively correlated with the protein levels of LC3B-II. TNF-alfa and IL-1Beta expression had also a positive correlation with protein level of p62. IL-10 mRNA expression was positively correlated with the mRNA expression of BECN1 and LAMP2 and protein levels of LC3B-II and negatively correlated with protein level of p62. In addition, p-mTOR level was positively correlated with IL-1Beta and TNF-alfa mRNA expression. The results revealed a reduced autophagy in PBMCs of T2D patients that is liked with an enhanced inflammation. The suppression of autophagy in PBMCs of diabetic patients may be associated with the activation of the mTOR signaling

Downregulation of lncRNA MALAT1 contributes to renal functional improvement after duodenal-jejunal bypass in a diabetic rat model

Ameliorated renal function has been reported after bariatric surgery, but the mechanisms underlying this phenomenon are not well-studied. To investigate whether the long non-coding RNA (lncRNA) MALAT1 mediates the amelioration of diabetic nephropathy after duodenal-jejunal bypass (DJB) surgery, rats were assigned randomly into four groups: diabetic (DM) group, DM with DJB surgery group, DM with sham surgery group, and healthy control group. Food intake, body weight, oral glucose tolerance test (OGTT), urine albumin excretion rate (UAER), and glomerular filtration rate (GFR) were measured and histological examination of renal sections was performed. For in vitro study, HK-2 cells were cultured under various glucose concentrations following MALAT1 siRNA transfection. Expression levels of MALAT1, SAA3, IL-6, and TNF-α in rat renal tissues or HK-2 cell lines were evaluated by qRT-PCR and/or ELISA. Results showed DJB surgery improved the renal function of diabetic rats, as indicated by ameliorated UAER and GFR and attenuated glomerular hypertrophy. Expression of MALAT1 and its downstream target SAA3 was significantly downregulated in renal tissues after DJB, which in turn decreased the expression of the pro-inflammatory cytokines IL-6 and TNF-α. Knockdown of MALAT1 in HK-2 cell lines further confirmed that expression levels of SAA3, IL-6, and TNF-alfa were regulated by MALAT1 under both low- and high-glucose conditions. Our findings suggest that MALAT1 is implicated in the improvement of renal function after DJB through regulation of its downstream targets SAA3, IL-6, and TNF-alfa

Physical training improves thermogenesis and insulin pathway, and induces remodeling in white and brown adipose tissues

Physical training (PT) has been considered as a treatment in metabolic syndrome (MS), since it induces thermogenic activity in brown (BAT) and white (WAT) adipose tissues. We evaluated the therapeutic effect of PT on activity of WAT and BAT in rats with MS induced by high-fat diet (30% lard) for 13 weeks and submitted, for the last 6 weeks, to swimming or kept sedentary (SED) rats. MS-SED rats compared to control diet (CT-SED) rats showed low physical fitness and high levels of glucose, insulin, homeostasis evaluation of insulin resistance (HOMA-IR), homeostasis evaluation of the functional capacity of Beta-cells (HOMA-Beta), and blood pressure. The gastrocnemius muscle decreased in peroxisome proliferator-activated receptor gamma coactivator 1-alpha and beta (PGC-1alfa, PGC-1beta), and uncoupled protein 2 and 3 (UCP2 and UCP3) expressions. Both WAT and BAT increased in the adipocyte area and decreased in blood vessels and fibroblast numbers. WAT increased in expression of pro-inflammatory adipokines and decreased in anti-inflammatory adipokine and adiponectin. WAT and gastrocnemius showed impairment in the insulin signaling pathway. In response to PT, MS rats showed increased physical fitness and restoration of certain biometric and biochemical parameters and blood pressure. PT also induced thermogenic modulations in skeletal muscle, WAT and BAT, and also improved the insulin signaling pathway. Collectively, PT was effective in treating MS by inducing improvement in physical fitness and interchangeable effects between skeletal muscle, WAT and BAT, suggesting a development of brown-like adipocyte cells

Voluntary exercise opposes insulin resistance of skeletal muscle glucose transport during liquid fructose ingestion in rats

We have recently reported that male rats given liquid fructose ingestion exhibit features of cardiometabolic abnormalities including non-obese insulin resistance with impaired insulin signaling transduction in skeletal muscle (Rattanavichit Y et al. Am J Physiol Regul Integr Comp Physiol 311: R1200-R1212, 2016). While exercise can attenuate obesity-related risks of cardiometabolic syndrome, the effectiveness and potential mechanism by which exercise modulates non-obese insulin resistance have not been fully studied. The present investigation evaluated whether regular exercise by voluntary wheel running (VWR) can reduce cardiometabolic risks induced by fructose ingestion. Moreover, the potential cellular adaptations following VWR on key signaling proteins known to influence insulin-induced glucose transport in skeletal muscle of fructose-ingested rats were investigated. Male Sprague-Dawley rats were given either water or liquid fructose (10% wt/vol) without or with access to running wheel for 6 weeks. We demonstrated that VWR restored insulin-stimulated glucose transport in the soleus muscle by improving the functionality of several signaling proteins, including insulin-stimulated IRBetaTyr1158/Tyr1162/Tyr1163 (82%), IRS-1 Tyr989 (112%), Akt Ser473 (56%), AS160 Thr642 (76%), and AS160 Ser588 (82%). These effects were accompanied by lower insulin-stimulated phosphorylation of IRS-1 Ser307 (37%) and JNK Thr183/Tyr185 (49%), without significant changes in expression of proteins in the renin-angiotensin system. Intriguingly, multiple cardiometabolic abnormalities were not observed in fructose-ingested rats with access to VWR. Collectively, this study demonstrates that the development of cardiometabolic abnormalities as well as insulin resistance of skeletal muscle and defective signaling molecules in rats induced by fructose ingestion could be opposed by VWR

Changes in the activity of some metabolic enzymes in the heart of SHR rat incurred by transgenic expression of CD36

Hypertension, dyslipidemia, and insulin resistance in the spontaneously hypertensive rat (SHR) can be alleviated by rescuing CD36 fatty acid translocase. The present study investigated whether transgenic rescue of CD36 in SHR could affect mitochondrial function and activity of selected metabolic enzymes in the heart. These analyses were conducted on ventricular preparations derived from SHR and from transgenic strain SHR-Cd36 that expresses a functional wild-type CD36. Our respirometric measurements revealed that mitochondria isolated from the left ventricles exhibited two times higher respiratory activity than those isolated from the right ventricles. Whereas, we did not observe any significant changes in functioning of the mitochondrial respiratory system between both rat strains, enzyme activities of total hexokinase, and both mitochondrial and total malate dehydrogenase were markedly decreased in the left ventricles of transgenic rats, compared to SHR. We also detected downregulated expression of the succinate dehydrogenase subunit SdhB (complex II) and 70 kDa peroxisomal membrane protein in the left ventricles of SHR-Cd36. These data indicate that CD36 may affect in a unique fashion metabolic substrate flexibility of the left and right ventricles

Clinical implications of the deregulated TP73 isoforms expression in cancer

TP73 is a member of the TP53 family whose expression has been observed altered in most human cancers and associated with the prognosis. TP73 translates into a complex number of isoforms with both oncogenic and tumour-suppressor functions and presents a complex cross-talk with other members of the family (TP53 and TP63). In this revision, we focus on the evidence that may support TP73 variants as prognostic markers in cancer. Nowadays, most publications in this topic highlight the association between overexpression of the oncogenic variants and failure to respond to chemotherapy and/or shorter survival. In addition, we comment on the putative possibilities that the detection through a liquid biopsy of TP73 variants may provide, and finally, the significance of determining the value of the combined alteration of the TP53 family members in the clinical setting

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Identification of miR-9 as a negative factor of insulin secretion from beta cells

MicroRNA is a novel class of small noncoding RNA that has been implicated in a variety of physiological and pathological processes, including glucose homeostasis and diabetes mellitus. So far, a few studies have reported that miRNAs may be an important regulator in glucose-stimulated insulin secretion (GSIS) pathway. However, the role of miRNAs in this process remains unclear. The levels of miRNAs in mouse islets and MIN6 cells were determined by quantitative RT-PCR. Concentration of insulin was determined by ELISA, and the expression of the target protein was determined with western blot assay. The overexpression and downregulation of miRNAs in MIN6 were conducted using cell transfection methods. And luciferase assay was used to measure the direct interaction between miRNAs and target messenger RNAs 3′UTR. miR-9 was screened out for it was downregulated under the effects of short-term high glucose, while long-term high glucose relatively increased miR-9 expression. The Stxbp1 expression was decreased with the overexpression of miR-9 in MIN6 cells and increased when miR-9 was downregulated. Moreover, it was verified by luciferase assay that miR-9 regulated Stxbp1 gene expression by directly targeting Stxbp1 messenger RNA 3′UTR. This study suggests that the pathway consisting of miR-9 and Stxbp1 plays a key role in β-cell function, thus contributing to the network of miRNA-insulin secretion and offering a new candidate for diabetes therapy