Blood Orange Juice Intake Modulates the Expression of miR-126–3p and let-7f-5p in PBMC of Overweight and Insulin Resistance Women

Abstract Objectives To investigate the effect of chronic blood orange juice intake on the microRNA profile in peripheral blood mononuclear cells (PBMC) of overweight and insulin resistance women. Methods Interventional and chronic study with women (n = 8) aged 18 to 40 years, diagnosed with overweight [body mass index (BMI) 25–29.9 kg/m2] and insulin resistance [homeostatic model assessment insulin resistance (HOMA-IR) index >2,71]. For four weeks, the volunteers ingested 500 mL/day of blood orange juice (Moro variety), with blood samples collected at baseline and four weeks after the beginning of drink ingestion. Evaluation of the expression of 137 microRNAs in PBMC was performed by real-time polymerase chain reaction (PCR). Results Blood orange juice intake decreased the expression of miR-126-3p (p = 0.004) and let-7f-5p (p = 0.005) in PBMC. These microRNAs are involved in suppressing the synthesis of several proteins of the insulin signaling pathway. Insulin receptor substrates (IRS) 1 and 2 were identified as target genes of mir-126. Insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), IRS2, phosphatidylinositol-3-kinase interacting protein 1 (PIK3IP1), and protein kinase B/Akt 2 (AKT2) were identified as target genes of let-7f. Conclusions Blood orange juice, rich in vitamin C, flavonoids, and anthocyanins, downregulates the expression of microRNA involved in impairing the insulin signaling pathway. Funding Sources Food Research Center (FoRC), São Paulo Research Foundation (FAPESP)

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Defective insulin signaling in skeletal muscle of the hypertensive TG(mREN2)27 rat

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00396.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. E1074-E1081 ◽

Cited By ~ 40

Author(s):

Julie A. Sloniger ◽

Vitoon Saengsirisuwan ◽

Cody J. Diehl ◽

Betsy B. Dokken ◽

Narissara Lailerd ◽

...

Keyword(s):

Insulin Resistance ◽

Skeletal Muscle ◽

Angiotensin Ii ◽

Insulin Receptor ◽

Signaling Pathway ◽

Glucose Transport ◽

Insulin Signaling ◽

Glycogen Synthase ◽

Insulin Signaling Pathway ◽

Whole Body

Essential hypertension is frequently associated with insulin resistance of skeletal muscle glucose transport, with a potential role of angiotensin II in the pathogenesis of both conditions. The male heterozygous TG(mREN2)27 rat harbors the mouse transgene for renin, exhibits local elevations in angiotensin II, and is an excellent model of both hypertension and insulin resistance. The present study was designed to investigate the potential cellular mechanisms for insulin resistance in this hypertensive animal model, including an assessment of elements of the insulin-signaling pathway. Compared with nontransgenic, normotensive Sprague-Dawley control rats, male heterozygous TG(mREN2)27 rats displayed elevated ( P < 0.05) fasting plasma insulin (74%), an exaggerated insulin response (108%) during an oral glucose tolerance test, and reduced whole body insulin sensitivity. TG(mREN2)27 rats also exhibited decreased insulin-mediated glucose transport and glycogen synthase activation in both the type IIb epitrochlearis (30 and 46%) and type I soleus (22 and 64%) muscles. Importantly, there were significant reductions (∼30–50%) in insulin stimulation of tyrosine phosphorylation of the insulin receptor β-subunit and insulin receptor substrate-1 (IRS-1), IRS-1 associated with the p85 subunit of phosphatidylinositol 3-kinase, Akt Ser473 phosphorylation, and Ser9 phosphorylation of glycogen synthase kinase-3β in epitrochlearis and soleus muscles of TG(mREN2)27 rats. Soleus muscle triglyceride concentration was 25% greater in the transgenic group compared with nontransgenic animals. Collectively, these data provide the first evidence that the insulin resistance of the hypertensive male heterozygous TG(mREN2)27 rat can be attributed to specific defects in the insulin-signaling pathway in skeletal muscle.

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MG53 is not a critical regulator of insulin signaling pathway in skeletal muscle

10.1101/2020.12.24.424288 ◽

2020 ◽

Author(s):

Clothilde Philouze ◽

Sophie Turban ◽

Béatrice Cremers ◽

Audrey Caliez ◽

Gwladys Lamarche ◽

...

Keyword(s):

Insulin Resistance ◽

Skeletal Muscle ◽

Insulin Receptor ◽

Signaling Pathway ◽

Insulin Signaling ◽

Insulin Response ◽

Muscle Cells ◽

Insulin Signaling Pathway ◽

Akt Phosphorylation ◽

Impaired Insulin

AbstractIn type 2 diabetes (T2D), both muscle and liver are severely resistant to insulin action. Muscle insulin resistance accounts for more than 80% of the impairment in total body glucose disposal in T2D patients and is often characterized by an impaired insulin signaling. Mitsugumin 53 (MG53), a muscle-specific TRIM family protein initially identified as a key regulator of cell membrane repair machinery has been suggested to be a critical regulator of muscle insulin signaling pathway by acting as ubiquitin E3 ligase targeting both the insulin receptor and insulin receptor substrate 1 (IRS1). Here, we show using in vitro and in vivo approaches that MG53 is not a critical regulator of insulin signaling and glucose homeostasis. First, MG53 expression is not consistently regulated in skeletal muscle from various preclinical models of insulin resistance. Second, MG53 gene knock-down in muscle cells does not lead to impaired insulin response as measured by Akt phosphorylation on Serine 473 and glucose uptake. Third, recombinant human MG53 does not alter insulin response in both differentiated C2C12 and human skeletal muscle cells. Fourth, ectopic expression of MG53 in HEK293 cells lacking endogenous MG53 expression fails to alter insulin response as measured by Akt phosphorylation. Finally, both male and female mg53 −/− mice were not resistant to high fat induced obesity and glucose intolerance compared to wild-type mice. Taken together, these results strongly suggest that MG53 is not a critical regulator of insulin signaling pathway in skeletal muscle.

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MG53 is not a critical regulator of insulin signaling pathway in skeletal muscle

PLoS ONE ◽

10.1371/journal.pone.0245179 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0245179

Author(s):

Clothilde Philouze ◽

Sophie Turban ◽

Beatrice Cremers ◽

Audrey Caliez ◽

Gwladys Lamarche ◽

...

Keyword(s):

Insulin Resistance ◽

Skeletal Muscle ◽

Insulin Receptor ◽

Signaling Pathway ◽

Insulin Signaling ◽

Insulin Response ◽

Muscle Cells ◽

Insulin Signaling Pathway ◽

Akt Phosphorylation ◽

Impaired Insulin

In type 2 diabetes (T2D), both muscle and liver are severely resistant to insulin action. Muscle insulin resistance accounts for more than 80% of the impairment in total body glucose disposal in T2D patients and is often characterized by an impaired insulin signaling. Mitsugumin 53 (MG53), a muscle-specific TRIM family protein initially identified as a key regulator of cell membrane repair machinery has been suggested to be a critical regulator of muscle insulin signaling pathway by acting as ubiquitin E3 ligase targeting both the insulin receptor and insulin receptor substrate 1 (IRS1). Here, we show using in vitro and in vivo approaches that MG53 is not a critical regulator of insulin signaling and glucose homeostasis. First, MG53 expression is not consistently regulated in skeletal muscle from various preclinical models of insulin resistance. Second, MG53 gene knock-down in muscle cells does not lead to impaired insulin response as measured by Akt phosphorylation on Serine 473 and glucose uptake. Third, recombinant human MG53 does not alter insulin response in both differentiated C2C12 and human skeletal muscle cells. Fourth, ectopic expression of MG53 in HEK293 cells lacking endogenous MG53 expression fails to alter insulin response as measured by Akt phosphorylation. Finally, both male and female mg53 -/- mice were not resistant to high fat induced obesity and glucose intolerance compared to wild-type mice. Taken together, these results strongly suggest that MG53 is not a critical regulator of insulin signaling pathway in skeletal muscle.

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Identification of transcription factors MYC and C/EBPβ mediated regulatory networks in heart failure based on GEO dataset

10.21203/rs.2.16967/v2 ◽

2020 ◽

Author(s):

Haiwei Wang ◽

Xinrui Wang ◽

Liangpu Xu ◽

Hua Cao

Keyword(s):

Heart Failure ◽

Transcription Factors ◽

Signaling Pathways ◽

Signaling Pathway ◽

Insulin Signaling ◽

Regulatory Networks ◽

Target Genes ◽

Transcriptional Profiling ◽

Insulin Signaling Pathway ◽

Failing Heart

Abstract Background: Heart failure is one of leading cause of death worldwide. However, the transcriptional profiling of heart failure is unclear. Moreover, the signaling pathways and transcription factors involving the heart failure development also are largely unknown. Using published Gene Expression Omnibus (GEO) datasets, in the present study, we aim to comprehensively analyze the differentially expressed genes in failing heart tissues, and identified the critical signaling pathways and transcription factors involving heart failure development. Methods: The transcriptional profiling of heart failure was identified from previously published gene expression datasets deposited in GSE5406, GSE16499 and GSE68316. The enriched signaling pathways and transcription factors were analyzed using DAVID website and gene set enrichment analysis (GSEA) assay. The transcriptional networks were created by Cytoscape. Results: Compared with the normal heart tissues, 90 genes were particularly differentially expressed in failing heart tissues, and those genes were associated with multiple metabolism signaling pathways and insulin signaling pathway. Metabolism and insulin signaling pathway were both inactivated in failing heart tissues. Transcription factors MYC and C/EBPβ were both negatively associated with the expression profiling of failing heart tissues in GSEA assay. Moreover, compared with normal heart tissues, MYC and C/EBPβ were down regulated in failing heart tissues. Furthermore, MYC and C/EBPβ mediated downstream target genes were also decreased in failing heart tissues. MYC and C/EBPβ were positively correlated with each other. At last, we constructed MYC and C/EBPβ mediated regulatory networks in failing heart tissues, and identified the MYC and C/EBPβ target genes which had been reported involving the heart failure developmental progress. Conclusions: Our results suggested that metabolism pathways and insulin signaling pathway, transcription factors MYC and C/EBPβ played critical roles in heart failure developmental progress.

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Altered Insulin Receptor Substrate 1 Phosphorylation in Blood Neuron-Derived Extracellular Vesicles From Patients With Parkinson’s Disease

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.564641 ◽

2020 ◽

Vol 8 ◽

Author(s):

Szu-Yi Chou ◽

Lung Chan ◽

Chen-Chih Chung ◽

Jing-Yuan Chiu ◽

Yi-Chen Hsieh ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Insulin Receptor ◽

Signaling Pathway ◽

Extracellular Vesicles ◽

Insulin Signaling ◽

Clinical Manifestations ◽

Insulin Signaling Pathway ◽

Insulin Receptor Substrate ◽

Neuronal Markers

IntroductionDiabetes increases the risk of Parkinson’s disease (PD). The phosphorylation of type 1 insulin receptor substrate (IRS-1) determines the function of insulin signaling pathway. Extracellular vesicles (EVs) are emerging as biomarkers of human diseases. The present study investigated whether PD patients exert altered phosphorylation IRS-1 (p-IRS-1) inside the blood neuron-derived extracellular vesicles (NDEVs).Research Design and MethodsIn total, there were 94 patients with PD and 63 healthy controls recruited and their clinical manifestations were evaluated. Blood NDEVs were isolated using the immunoprecipitation method, and Western blot analysis was conducted to assess total IRS-1, p-IRS-1, and downstream substrates level in blood NDEVs. Statistical analysis was performed using SPSS 19.0, and p < 0.05 was considered significant.ResultsThe isolated blood EVs were validated according to the presence of CD63 and HSP70, nanoparticle tracking analysis and transmission electron microscopy. NDEVs were positive with neuronal markers. PD patients exerted significantly higher level of p-IRS-1S312 in blood NDEVs than controls. In addition, the p-IRS-1S312 levels in blood NDEVs was positively associated with the severity of tremor in PD patients after adjusting of age, sex, hemoglobin A1c, and body mass index (BMI).ConclusionPD patients exerted altered p-IRS-1S312 in the blood NDEVs, and also correlated with the severity of tremor. These findings suggested the association between dysfunctional insulin signaling pathway with PD. The role of altered p-IRS-1S312 in blood NDEVs as a segregating biomarker of PD required further cohort study to assess the association with the progression of PD.

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268 THE EFFECTS OF METFORMIN ON THE INSULIN SIGNALING PATHWAY ON PORCINE GRANULOSA LUTEIN CELLS IN A PRIMARY CULTURE SYSTEM

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab268 ◽

2007 ◽

Vol 19 (1) ◽

pp. 250

Author(s):

M. S. Hossein ◽

M. S. Lee ◽

W. S. Hwang

Keyword(s):

Insulin Receptor ◽

Signaling Pathway ◽

Granulosa Cells ◽

Insulin Signaling ◽

Luciferase Activity ◽

Insulin Signaling Pathway ◽

Nuclear Factor ◽

Activator Protein 1 ◽

Activator Protein ◽

Nuclear Factor Kb

Metformin (N,N22-dimethylbiguanide) is an oral antihyperglycaemic drug which increases insulin-stimulated glucose uptake as an insulin sensitizing agent (ISA) and has direct effects on ovarian steroidogenesis in human. To investigate the effects of metformin on the insulin signaling pathway on porcine granulosa lutein cells in a primary culture system, we examined mRNA expressions of porcine insulin receptor, insulin-like growth factor-1 receptor, and insulin receptor substrate-1; expressions of downstream targets (Raf, MEK1/2, ERK, PDK1, mTOR, p70, and nuclear factor kB) of the insulin receptor signaling pathway; the luciferase activity of transcription factors activator protein 1; and nuclear factor kB. Granulosa cells were plated in DMEM and 10% fetal bovine serum at a density of 3.5 � 107 in a T-25 tissue culture flask and cultured for 48 h at 37�C in a humidified atmosphere of 5% CO2 and 95% air. Then, porcine granulosa lutein cells (pGLs) were cultured in a serum-free DMEM as a control group or supplemented with 10-5 M metformin, 100 ng mL-1 of insulin, or both for 24 h. The monolayer was collected for RT-PCR and western blot analysis. For transient transfections, porcine granulosa cells were plated in DMEM and 10% FBS at a concentration of 2.5 � 106/well in 24-well culture dishes for 16 h and then transfected with plasmid constructs using FuGene6 Transfection Reagent. Reporter vectors used in these studies were AP-1-luc and NF-κ B-luc. Cells were treated post-transfection with 10-5 M metformin, 100 ng mL-1 insulin, or both for 48 h. The luciferase activity was assayed using the dual luciferase assay kit with a Microlumat LB 96 P luminometer. Metformin with insulin significantly increased mRNA expressions of insulin receptor, insulin-like growth factor-1 receptor, and insulin receptor substrate-1, whereas metformin alone had no significant effect on the expression of genes. Metformin alone and with insulin significantly increased expressions of downstream targets of the insulin receptor signaling pathway except MEK2. Metformin with insulin significantly elicited an induction of luciferase activity in the transfection of activator protein 1 and nuclear factor kB reporter, whereas metformin alone did not. Metformin induced expressions of target molecules in the insulin signaling pathway but had no effect on the luciferase activity of activator protein 1 and nuclear factor kB in porcine granulosa lutein cells. Metformin with insulin increased the luciferase activity of activator protein 1. These results suggest that metformin with insulin could change the function of ovarian granulosa cells induced by activator protein 1 activity.

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Sphingomyelinase has an insulin-like effect on glucose transporter translocation in adipocytes

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1998.274.5.r1446 ◽

1998 ◽

Vol 274 (5) ◽

pp. R1446-R1453 ◽

Cited By ~ 2

Author(s):

T. S. David ◽

P. A. Ortiz ◽

T. R. Smith ◽

J. Turinsky

Keyword(s):

Plasma Membrane ◽

Insulin Receptor ◽

Glucose Uptake ◽

Signaling Pathway ◽

Insulin Signaling ◽

Glucose Transporter ◽

Insulin Signaling Pathway ◽

Glut 1 ◽

Glut 4 ◽

Glut 4 Translocation

Rat epididymal adipocytes were incubated with 0, 0.1, and 1 mU sphingomyelinase/ml for 30 or 60 min, and glucose uptake and GLUT-1 and GLUT-4 translocation were assessed. Adipocytes exposed to 1 mU sphingomyelinase/ml exhibited a 173% increase in glucose uptake. Sphingomyelinase had no effect on the abundance of GLUT-1 in the plasma membrane of adipocytes. In contrast, 1 mU sphingomyelinase/ml increased plasma membrane content of GLUT-4 by 120% and produced a simultaneous decrease in GLUT-4 abundance in the low-density microsomal fraction. Sphingomyelinase had no effect on tyrosine phosphorylation of either the insulin receptor β-subunit or the insulin receptor substrate-1, a signaling molecule in the insulin signaling pathway. It is concluded that the incubation of adipocytes with sphingomyelinase results in insulin-like translocation of GLUT-4 to the plasma membrane and that this translocation does not occur via the activation of the initial components of the insulin signaling pathway.

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