Glucose Uptake Activity Research Articles

192-OR: A CRISPR Screen Identifies Rfwd2 as a Regulator of Brown Fat Glucose Transport

Brown adipose tissue (BAT) generates heat to regulate body temperature, and to support this energy expending activity it takes up glucose from circulation. Cold exposure regimens that activate BAT improve insulin sensitivity, presumably by decreasing circulating glucose levels. Interestingly, glucose uptake during cold is specific to BAT and occurs at low circulating insulin levels, suggesting alternative pathways that do not exist in other insulin sensitive tissues may regulate brown adipocyte glucose uptake. To identify putative regulators of brown adipocyte glucose uptake, we performed a whole genome CRISPR screen using the uptake of the fluorescent glucose analog 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) as the readout on mouse brown adipocytes differentiated in vitro. By sorting single brown adipocytes according to their fluorescence, we separated the top 5%, bottom 5% and the remainder of cells using fluorescence activated cell sorting (FACS) and then sequenced to identify genes acting as both putative positive and negative regulators of glucose uptake. Genes identified as potential hits could be classified by gene ontology and belonged mostly to groups related to metabolism and energy homeostasis. A list of 25 potential regulatory genes were validated using the same 2-NBDG strategy used to identify them as well as radiolabeled glucose uptake in vitro. From these genes, 5 were further validated using a second CRISPR based strategy to block gene function. One gene that we identified is the E3 ubiquitin ligase Rfwd2, which is highly expressed in BAT and is known to regulate lipid metabolism. When Rfwd2 was deleted from brown adipocytes the global ubiquitination pattern was changed and glucose uptake was increased in both the basal and insulin stimulated states. These data suggest that targets of this ligase may play a role in the cell-signaling cascade that activates glucose uptake and these targets could further regulate BAT glucose uptake in vivo. Disclosure M. Lynes: None. Q. Huang: None. P. Yi: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd. Funding National Institutes of Health (P30DK036836, R01DK077097, R01DK102898, F32DK102320, K01DK111714)

Spectral tracing of deuterium for imaging glucose metabolism.

Cells and tissues often display pronounced spatial and dynamical metabolic heterogeneity. Prevalent glucose-imaging techniques report glucose uptake or catabolism activity, yet do not trace the functional utilization of glucose-derived anabolic products. Here, we report a microscopy technique for the optical imaging, via the spectral tracing of deuterium (referred to as STRIDE), of diverse macromolecules derived from glucose. Based on stimulated-Raman-scattering imaging, STRIDE visualizes the metabolic dynamics of newly synthesized macromolecules, such as DNA, protein, lipids and glycogen, via the enrichment and distinct spectra of carbon–deuterium bonds transferred from the deuterated glucose precursor. STRIDE can also use spectral differences derived from different glucose isotopologues to visualize temporally separated glucose populations in a pulse–chase manner. We also show that STRIDE can be used to image glucose metabolism in many mouse tissues, including tumours, the brain, the intestine and the liver, at a detection limit of 10 mM of carbon–deuterium bonds. STRIDE provides a high-resolution and chemically informative assessment of glucose anabolic utilization.

Fibroblast growth factor 21 secretion enhances glucose uptake in mono(2-ethylhexyl)phthalate-treated adipocytes

Previous studies revealed that cellular accumulation of mono(2-ethylhexyl)phthalate (MEHP) disturbed energy metabolism in adipocytes, where glucose uptake was significantly increased. The present study aimed to determine the mechanisms underlying the increased glucose uptake. MEHP-treated 3T3-L1 adipocytes exhibited a significantly increased glucose uptake activity. Immunoblot analysis suggested that the insulin-induced signals were not responsible for the increased glucose uptake. qPCR analysis revealed that both Glut1 and Glut4 genes were highly expressed during adipogenesis; Glut1 mRNA levels in MEHP-treated adipocytes were significantly increased. Moreover, MEHP-treated adipocytes exhibited significantly increased levels of fibroblast growth factor 21 (FGF21) in both mRNA and secreted protein. FGF21 is a peptide hormone with pleiotropic effects on regulation of insulin sensitivity and glucose/lipid homeostasis. We found that MEHP, FGF21, and lactate in culture medium together enhanced Fgf21 gene expression in MEHP-treated adipocytes. FGF21 signaling requires fibroblast growth factor receptor (FGFR) and βKlotho. Fgfr family and βKlotho genes were actively expressed during adipogenesis; mRNA levels of Fgfr3 and Fgfr4 genes in MEHP-treated adipocytes were significantly increased. Roles of FGF21/FGFR and phosphoinositide 3-kinase (PI3K)/AKT signal axes in regulation of glucose uptake were determined. We demonstrated that FGF21/FGFR signals played the major roles in up-regulation of the basal glucose uptake in MEHP-treated adipocytes. The in vitro evidence suggests that cellular FGF21 secretion enhances the basal glucose uptake in MEHP-treated adipocytes.

SUN-080 Diminished Akt Activation and Interaction with 14-3-3ζ is Associated with Insulin Resistance in Cardiomyocytes of Metabolic Syndrome Rats

Metabolic syndrome (MetS) is a cluster of physiological and biochemical disorders that increases the risk of cardiovascular disease, being insulin resistance (IR) a pivotal condition for its development. Alterations such as metabolic inflexibility, changes in calcium dynamics, oxidative stress and impaired insulin signaling have been reported in the state of IR in the heart. Akt kinase activation plays a central role in insulin actions, leading to glucose uptake and its use as the primary energy source in the heart. Akt activity and subcellular localization are highly regulated by phosphorylation and by the interaction with other proteins. It has been reported that 14-3-3 proteins regulate insulin response and in particular the ζ isoform exerts a positive regulation on Akt activation. Therefore, we studied the changes in Akt activation and interaction with 14-3-3ζ, in response to insulin in cardiomyocytes from a MetS rat model, induced by the consumption of 30% sucrose in the drinking water during 4 months. MetS rats developed obesity (body weight in g: MetS: 592±12 vs 415±5 in control rats (C), P<0.001; epididymal fat in g: MetS: 17.5±1 vs C: 7.3±0.4, P<0.001), hypertriglyceridemia (in mg/dL MetS: 137±10 vs C: 60±3, P<0.001), low HDL-cholesterol levels (in mg/dL MetS: 31±2 vs C: 44±2, P<0.01), and systemic insulin resistance (intraperitoneal glucose tolerance test, area under the curve: MetS: 34199±2555 vs C: 17908±1844, P<0.001), without significant modifications in fasting blood glucose levels (in mg/dL MetS: 76±6 vs C: 65±5, P=0.17). Isolated cardiomyocytes from MetS rats showed insulin resistance, characterized by the absence of glucose uptake after stimulation with this hormone (3H-2deoxyglucose uptake in fmol/mg of protein/min: MetS basal: 84±17 vs MetS+insulin: 94±20, P=0.63; C basal: 74±19 vs C+insulin: 190±47, P<0.05). Insulin-induced Akt phosphorylation (determined by Western Blot, after 10 min of stimulation with 100 nM) in the key activation residues Ser473/Thr308 was diminished by 51% and 66%, respectively. Insulin-induced Akt translocation towards the plasma membrane (determined by immunostaining/confocal microscopy) a process involved in its activation, was also decreased in MetS cardiomyocytes. On the other hand, we found a diminished insulin-induced Akt-14-3-3ζ interaction evaluated by Akt co-immunoprecipitation/Western blot and by Akt co-immunoprecipitation/label-free mass spectrometry, since 14-3-3 was exclusively found in a pool of 3 C rats cardiomyocytes samples, but not in the MetS pool; this impairment suggest a reduced activation of AS160 protein and glucose uptake. These findings unveil a key participation of Akt regulatory mechanisms in the establishment of IR in cardiomyocytes under the MetS condition. Sources of Research Support: PRODEP research grant to the Academic Group Cinvestav-CA-10 to AR and JAO-R. CONACYT scholarship 278067 to HVL-G.

IL‐15 Upregulates GLUT1 while Reducing CPT1 in Cardiomyocytes

Obesity is an exponentially growing epidemic in the United States and is a leading contributor of cardiovascular disease (CVD). Maintenance of lifestyle interventions to prevent and treat obesity, such as diet and exercise, is a challenge. In this regard, targeting the molecular mediators of obesity has become a focus to prevent CVD disease progression. In an undiseased state, the cardiac muscle relies primarily on oxidative processes for energy production. In some cases, CVD can lead to upregulation of glycogenic, rather than oxidative, processes to produce ATP for muscle contraction. The myokine interleukin‐15 (IL‐15) is secreted by skeletal muscle during exercise and has been shown to reduce obesity and related disorders. However, the effects of IL‐15 signaling in cardiomyocytes (CM) remains largely unknown. The objective of the current study was to determine if IL‐15 regulates glucose and/or fatty acid (FA) metabolism in CM. Differentiated CM were treated with 100 ng/mL of IL‐15 for 24 hours. Protein was harvested, from either IL‐15 or vehicle treated cells, and isolated into total, mitochondrial and plasma membrane (PM) fractions. The protein concentration was quantified utilizing a Bradford assay. Subsequently, protein analysis, via Western blotting, was performed. Fifteen micrograms of protein was loaded onto 4–20% polyacrylamide gels for electrophoresis. The proteins were then transferred onto a PVDF membrane. Primary antibodies against GLUT1, GLUT4, and CPT1 (1:1000) were used, followed by secondary antibodies (1:2000). GLUT1 and GLUT4 were analyzed in both total cell fractions and PM fractions, while CPT1 levels were assessed in mitochondrial fractions. The membranes were subjected to enhanced chemiluminescence, imaged and quantified with ImageJ software. A t‐test (P&lt;0.05) was used to determine if IL‐15 had any significant effects on the protein levels of the aforementioned proteins. IL‐15 did not alter total GLUT4, PM GLUT4 or PM GLUT1 (P&gt;0.05). Conversely, IL‐15 treatment resulted in a 367% increase in total GLUT1 protein levels when compared to the vehicle control cells (P&lt;0.05). Of note, IL‐15 decreased mitochondrial CPT1 by nearly 50% (P&lt;0.05). Our data indicate that IL‐15 increases the protein concentration of one of the glucose transporters, GLUT1, and decreases the protein concentration of the mitochondrial FA transporter, CPT1, in CM. Thus, IL‐15 may be responsible for promoting a shift in CM metabolism from FA to glucose utilization. Additional studies are warranted to confirm this proposed shift in metabolism by studying glucose uptake and beta‐oxidation activity in CM. Further, to understand if an IL‐15‐mediated shift in metabolism is beneficial to CM, further studies aimed at assessing cardiac muscle function are required. Taken together, if IL‐15 holds the potential as a treatment for obesity, examination of its functions in various tissues is required.Support or Funding InformationChapman University Crean College of Health and Behavioral Sciences Summer Undergraduate Research Fellowship; American Heart Association 16SDG30680003This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Metabolic gatekeepers to safeguard against autoimmunity and oncogenic B cell transformation.

B cells face multiple restrictions on glucose and energy metabolism. Their lineage-determining transcription factors repress glucose uptake and pentose phosphate pathway activity, while their low numbers of mitochondria and small cytoplasmic volume set narrow limits for mitochondrial ATP production and autophagy as alternative energy sources. During activation, B cells can balance temporary increases of energy expenditure. However, permanent hyperactivation of kinases, for instance, downstream of an autoreactive B cell receptor (BCR) or a transforming oncogene, can cause energy stress and cell death. Here, I propose that B cell-intrinsic restriction of ATP represents a safeguard to eliminate autoreactive or pre-malignant B cells. If the metabolic gatekeepers are compromised, influx of additional glucose may fuel permanent increases in metabolic demands and pathological B cell proliferation, driven by an autoreactive BCR or a transforming oncogene.

Targeting the Zinc Transporter ZIP7 in the Treatment of Insulin Resistance and Type 2 Diabetes.

Type 2 diabetes mellitus (T2DM) is a disease associated with dysfunctional metabolic processes that lead to abnormally high levels of blood glucose. Preceding the development of T2DM is insulin resistance (IR), a disorder associated with suppressed or delayed responses to insulin. The effects of this response are predominately mediated through aberrant cell signalling processes and compromised glucose uptake into peripheral tissue including adipose, liver and skeletal muscle. Moreover, a major factor considered to be the cause of IR is endoplasmic reticulum (ER) stress. This subcellular organelle plays a pivotal role in protein folding and processes that increase ER stress, leads to maladaptive responses that result in cell death. Recently, zinc and the proteins that transport this metal ion have been implicated in the ER stress response. Specifically, the ER-specific zinc transporter ZIP7, coined the “gate-keeper” of zinc release from the ER into the cytosol, was shown to be essential for maintaining ER homeostasis in intestinal epithelium and myeloid leukaemia cells. Moreover, ZIP7 controls essential cell signalling pathways similar to insulin and activates glucose uptake in skeletal muscle. Accordingly, ZIP7 may be essential for the control of ER localized zinc and mechanisms that disrupt this process may lead to ER-stress and contribute to IR. Accordingly, understanding the mechanisms of ZIP7 action in the context of IR may provide opportunities to develop novel therapeutic options to target this transporter in the treatment of IR and subsequent T2DM.

Two new compounds of Penicillium polonicum, an endophytic fungus from Camptotheca acuminata Decne

To discover novel structural compounds which are producted by endophytic fungi, a primary chemical profiling of Camptotheca acuminata Decne derived endophytic fungus Penicillum polonicum had been taken. Two new compounds β-lactone polonicin A (1) and enoic acid polonicin B (2) together with seven known compounds 3-9 were isolated from Penicillum polonicum obtained from C. acuminata. The structures of the new compounds 1 and 2 were identified by modern spectrum technology including detailed 1D, 2D NMR and MS data analyses. When tested against HepG2 hepatocellular carcinoma (HCC) cell lines, compounds 4-8 showed moderate anti-HCC activity. In addition, compound 1-3 have effects on increasing GLUT4 translocation and glucose uptake in vitro. Compound 1 showed the strongest glucose uptake and GLUT4 translocation activities in rat skeleton (L6) myoblast cell line with enhancements of 1.8 and 2.1 folds respectively compared to the control.

Invitro Cytotoxicity and Glucose Uptake Activity of Mushroom Pleurotus Eous in Methanol Extract Using L-6 Cell Line

The aim of the present study is to evaluate the cytotoxicity and glucose uptake activity of methanolic extract of pleurotus eous mushroom using L6 cell lines. An MTT assay is a colorimetric assay based on assessing the cell metabolic activityThe results showed that the extracts did not confer any cytotoxicity. The effects were observed in treated cells up to 6.25 mg/ml extract as measured with MTT. Also the mushroom extracts shows better glucose uptake potential. The results were compared with Insulin a standard antidiabetic drug. Insulin enhance the glucose uptake upto 97.8%. The L6 cell lines enhance the glucose uptake by 57.4% at 65µg concentration.

TBC1D8 Amplification Drives Tumorigenesis through Metabolism Reprogramming in Ovarian Cancer.

Cancer cells undergo metabolic reprogramming to support their energy demand and biomass synthesis. However, the mechanisms driving cancer metabolism reprogramming are not well understood.Methods: The differential proteins and interacted proteins were identified by proteomics. Western blot, qRT-PCR and IHC staining were used to analyze TBC1D8 levels. In vivo tumorigenesis and metastasis were performed by xenograft tumor model. Cross-Linking assays were designed to analyze PKM2 polymerization. Lactate production, glucose uptake and PK activity were determined.Results: We established two aggressive ovarian cancer (OVCA) cell models with increased aerobic glycolysis. TBC1D8, a member of the TBC domain protein family, was significantly up-regulated in the more aggressive OVCA cells. TBC1D8 is amplified and up-regulated in OVCA tissues. OVCA patients with high TBC1D8 levels have poorer prognoses. TBC1D8 promotes OVCA tumorigenesis and aerobic glycolysis in a GAP activity-independent manner in vitro and in vivo. TBC1D8 bound to PKM2, not PKM1, via its Rab-GAP TBC domain. Mechanistically, TBC1D8 binds to PKM2 and hinders PKM2 tetramerization to decreases pyruvate kinase activity and promote aerobic glycolysis, and to promote the nuclear translocation of PKM2, which induces the expression of genes which are involved in glucose metabolism and cell cycle.Conclusions: TBC1D8 drives OVCA tumorigenesis and metabolic reprogramming, and TBC1D8 serves as an independent prognosis factor for OVCA patients.

A study of In-vitro hypoglycemic and glucose uptake activity of isolated compound from ethanolic leaf extract of Amaranthus tristis Linn.

Background: Natural bioactive compounds have a long history to use effectively in the treatment of Diabetes Mellitus. Amaranthus tristis Linn (Amaranthaceae) have been used for the treatment of various ailments. Objective: The present work was undertaken to study the effect of an isolated bioactive compound from the leaf of Amaranthus tristis Linn on glucose uptake in cell lines like 3T3-L1 cell lines and inhibition of alpha-glucosidase and alpha-amylase enzymes. Materials and Methods: The Ethanolic leaf extract of Amaranthus tristis Linn subjected to preliminary phytochemical screening and isolated compound were tested for its cytotoxicity study by MTT assay. Isolated compound was selected further glucose uptake assay based on cytotoxicity concentration CTC50 value. NMR technique was used to identify the structure of the compound. Results: Phytochemical analysis showed the presence of alkaloids, flavonoids, glycosides. The Isolated compound showed moderate cytotoxic activity to 3T3-L1 cell line. Compounds identified as rutin In in-vitro glucose uptake assay of rutin showed an increase in glucose uptake. Conclusion: A. tristis Linn. showed significant antidiabetic activity in dose-dependent manner. Abbreviations Used: MTT: 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, FBS: Fetal Bovine serum, PBS buffer: Phosphate Buffered Saline, DMEM: Modified Eagle's Medium, EDTA: Ethylenediaminetetraacetic acid, IBMX: 3-isobutyl-1-methylxanthine, TLC: Thin-layer chromatography, FTIR: Fourier-transform infrared spectroscopy, 13CNMR: nuclear magnetic resonance, 1H NMR: Proton nuclear magnetic resonance, GC-MS: Gas chromatography–mass spectrometry, DNSA: Dinitro salicylic acid.

Antidiabetic Activity of Zingiber officinale Roscoe Rhizome Extract: an In Vitro Study

The potential roles of Zingiber officinale Roscoe (ginger) for treating and preventing diabetes have been investigated in both humans and experimental animals. However, the mode of its action has not yet been elucidated. This study aimed to investigate the effects of ginger extract on glucose uptake activity and its activation pathway in L6 myotubes. Cells were co-cultured for 24 h with a variable concentration of either ginger extract or 2 mM metformin or 200 nM insulin or 20 μM Troglitazone (TGZ), followed by a 10-min 2-[3H]-deoxy-D-glucose (2-DG) uptake. The levels of glucose transporters 1 (GLUT1) and GLUT4 protein and mRNA expression were determined. Ginger extract at 400 μg/ml significantly enhanced glucose uptake in L6 myotubes (208.03 ± 10.65% above basal value, p&lt;0.05) after co-culture for 24 h. The ginger-enhancement of glucose uptake was inhibited by 3.5 μM cycloheximide, a protein synthesis inhibitor, 1 μM wortmannin (Phosphatidylinositol 3-Kinase (PI3 kinase) inhibitor) and 15 nM rapamycin (mammalian target of rapamycin (mTOR) inhibitor). The enhancement of glucose transport by ginger extract at 400 μg/ml was accompanied with the increased expression of GLUT1 protein (1.60 ± 0.20, 2.03 ± 0.19, and 2.25 ± 0.35 folds of basal at 4, 8, and 24 h, respectively p&lt;0.05) and mRNA (1.22 ± 0.96, 1.45 ± 0.93, 1.91 ± 0.75, 2.32±0.92, and 2.20 ± 0.64 folds of basal at 1, 2, 4, 8, and 24 h, respectively p&lt;0.05) in a time-dependent manner. Z. officinale Roscoe rhizome extract increase glucose transport activity of L6 myotubes by enhancing GLUT1 expression, the results of PI3-Kinase and 5’-AMP-activated kinase (AMPK) stimulation.

Novel glitazones as PPAR\u03b3 agonists: molecular design, synthesis, glucose uptake activity and 3D QSAR studies

BackgroundAn alarming requirement for finding newer antidiabetic glitazones as agonists to PPARγ are on its utmost need from past few years as the side effects associated with the available drug therapy is dreadful. In this context, herein, we have made an attempt to develop some novel glitazones as PPARγ agonists, by rational and computer aided drug design approach by implementing the principles of bioisosterism. The designed glitazones are scored for similarity with the developed 3D pharmacophore model and subjected for docking studies against PPARγ proteins. Synthesized by adopting appropriate synthetic methodology and evaluated for in vitro cytotoxicity and glucose uptake assay. Illustrations about the molecular design of glitazones, synthesis, analysis, glucose uptake activity and SAR via 3D QSAR studies are reported.ResultsThe computationally designed and synthesized ligands such as 2-(4-((substituted phenylimino)methyl)phenoxy)acetic acid derivatives were analysed by IR, 1H-NMR, 13C-NMR and MS-spectral techniques. The synthesized compounds were evaluated for their in vitro cytotoxicity and glucose uptake assay on 3T3-L1 and L6 cells. Further the activity data was used to develop 3D QSAR model to establish structure activity relationships for glucose uptake activity via CoMSIA studies.ConclusionThe results of pharmacophore, molecular docking study and in vitro evaluation of synthesized compounds were found to be in good correlation. Specifically, CPD03, 07, 08, 18, 19, 21 and 24 are the candidate glitazones exhibited significant glucose uptake activity. 3D-QSAR model revealed the scope for possible further modifications as part of optimisation to find potent anti-diabetic agents.

Antidiabetic potential of Musa spp. inflorescence: a systematic review.

Extracts of parts Musa spp. have been used for the treatment of various diseases in traditional medicine. Studies have shown that these extracts have hypoglycaemic properties. The aim of this work was to gather evidence on the antidiabetic effects of Musa spp. inflorescence. A systematic review was conducted with searches in three electronic databases, along with manual searches. Studies evaluating the antidiabetic properties of extracts of flower or bract of the genus Musa (in vitro or in vivo) were included. Overall, 16 studies were found. The reported assays were of hypoglycaemic effects, oral glucose tolerance, inhibitory activities in carbohydrate metabolism and digestive enzymes, enhanced glucose uptake activity and popular use of the extract in patients with diabetes type 2. In vitro studies showed that use of the extract was associated with antidiabetic effects (e.g. increased glucose uptake and inhibition of carbohydrate digestion enzymes). In induced diabetic models, Musa spp. extracts showed dose-dependent glycaemic level reductions compared with pharmacological drugs (P < 0.05). In general, promising results regarding antidiabetic activity were found for inflorescence of Musa spp., suggesting that this plant could represent a natural alternative therapy for treating diabetes mellitus type 2.

Timbe (Acaciella angustissima) Pods Extracts Reduce the Levels of Glucose, Insulin and Improved Physiological Parameters, Hypolipidemic Effect, Oxidative Stress and Renal Damage in Streptozotocin-Induced Diabetic Rats.

In Mexico one in 14 deaths are caused by diabetes mellitus (DM) or by the macro and microvascular disorders derived from it. A continuous hyperglycemic state is characteristic of DM, resulting from a sustained state of insulin resistance and/or a dysfunction of β-pancreatic cells. Acaciella angustissima is a little studied species showing a significant antioxidant activity that can be used as treatment of this disease or preventive against the complications. The objective of this study was to explore the effect of oral administration of A. angustissima methanol extract on physiological parameters of streptozotocin-induced diabetic rats. The results indicated a significant reduction in blood glucose levels, an increase in serum insulin concentration, a decrease in lipid levels and an improvement in the parameters of kidney damage by applying a concentration of 100 mg/Kg B.W. However, glucose uptake activity was not observed in the adipocyte assay. Moreover, the extract of A. angustissima displayed potential for the complementary treatment of diabetes and its complications likely due to the presence of bioactive compounds such as protocatechuic acid. This study demonstrated that methanol extract of Acacciella angustissima has an antidiabetic effect by reducing the levels of glucose, insulin and improved physiological parameters, hypolipidemic effect, oxidative stress and renal damage in diabetic rats.

Cell line cytotoxicity, antiadipogenic and glucose uptake activity of Sarcostemma brevistigma Wight. & Arn.

Nature has gifted us with abundant plants possessing medicinal virtues which can cure several illnesses. Currently, diabetes mellitus is a severe threat to human well-being across the world due to the rapidly increasing incidence of diabetes. New effective bioactive drugs are in need, as plants do harbour and are proven to have potential antidiabetic activity than the present hypoglycemic medicines used in clinical therapy. In this study, in vitro cytotoxicity, glucose uptake, and anti-adipogenic activities of the plant extract (methanolic extract) of S. brevistigma were examined using 3T3L1 cell lines. The studies interpreted by MTT cytotoxicity assay and glucose uptake assay by using 3T3-L1 cell lines, it was found that a very low dosage (1ng/mL) of plant extract showed lesser cytotoxicity effect (1.42%) and considerably higher glucose uptake activity of 38.04% which is equivalent to the glucose uptake shown by 100nm insulin (40.10%). Though plant extract has antidiabetic activity, it is important to study whether it has any other related side effects when used at higher concentrations. Therefore, in this study, the appropriate non-toxic concentration was optimized.

Excess Hydrocortisone Hampers Placental Nutrient Uptake Disrupting Cellular Metabolism.

Low birth weight increases neonatal morbidity and mortality, and surviving infants have increased risk of metabolic and cardiovascular disturbances later in life, as well as other neurological, psychiatric, and immune complications. A gestational excess of glucocorticoids (GCs) is a well-known cause for fetal growth retardation, but the biological basis for this association remains elusive. Placental growth is closely related to fetal growth. The placenta is the main regulator of nutrient transport to the fetus, resulting from the difference between placental nutrient uptake and the placenta's own metabolism. The aim of this study was to analyze how excess hydrocortisone affects placental glucose and lipid metabolism. Human placenta explants from term physiological pregnancies were cultured for 18 hours under different hydrocortisone concentrations (2.75, 5.5, and 55 mM; 1, 2, and 20 mg/ml). Placental glucose and lipid uptake and the metabolic partitioning of fatty acids were quantified by isotopic techniques, and expression of specific glucose transporter GLUT1 was quantified by western blot. Cell viability was assessed by MTT, immunohistochemistry and caspase activity. We found that excess hydrocortisone impairs glucose uptake and lipoprotein lipase (LPL) activity, coincident with a GC-dose dependent inhibition of fatty acid oxidation and esterification. None of the experimental conditions showed an increased cell death. In conclusion, our results show that GC overexposure exerts a dysfunctional effect on lipid transport and metabolism and glucose uptake in human placental explants. These findings could well be directly related to a reduced placental growth and possibly to a reduced supply of nutrients to the fetus and the consequent fetal growth retardation and metabolic programming.

Mitochondrial dysfunction and increased glycolysis in prodromal and early Parkinson's blood cells.

ABSTRACT Background: Although primarily a neurodegenerative process, there is increasing awareness of peripheral disease mechanisms in Parkinson's disease. To investigate disease processes in accessible patient cells, we studied peripheral blood mononuclear cells in recently diagnosed PD patients and rapid eye movement‐sleep behavior disorder patients who have a greatly increased risk of developing PD. We hypothesized that peripheral blood mononuclear cells may recapitulate cellular pathology found in the PD brain and investigated these cells for mitochondrial dysfunction and oxidative stress. Methods: Peripheral blood mononuclear cells were isolated and studied from PD patients, rapid eye movement‐sleep behavior disorder patients and age‐ and sex‐matched control individuals from the well‐characterized Oxford Discovery cohort. All participants underwent thorough clinical assessment. Results: Initial characterization showed that PD patients had elevated levels of CD14 + monocytes and monocytes expressing C‐C motif chemokine receptor 2. Mitochondrial dysfunction and oxidative stress were increased in PD patient peripheral blood mononuclear cells, with elevated levels of mitochondrial reactive oxygen species specifically in patient monocytes. This was combined with reduced levels of the antioxidant superoxide dismutase in blood cells from PD patients and, importantly, also in rapid eye movement‐sleep behavior disorder patients. This mitochondrial dysfunction was associated with a concomitant increase in glycolysis in both PD and rapid eye movement‐sleep behavior disorder patient blood cells independent of glucose uptake or monocyte activation. Conclusions: This work demonstrates functional bioenergetic deficits in PD and rapid eye movement‐sleep behavior disorder patient blood cells during the early stages of human disease. © 2018 The Authors. Movement Disorders published by Wiley Periodicals, Inc. on behalf of International Parkinson and Movement Disorder Society.

STAT3/c-Myc Axis-Mediated Metabolism Alternations of Inflammation-Related Glycolysis Involve with Colorectal Carcinogenesis.

Chronic inflammation is a major driving factor for the development of colitis-associated cancer (CAC). It is extensively acknowledged that patients who have long-standing inflammation bowel disease are at high risk for developing CAC. However, the metabolic alteration by which chronic intestinal inflammation promotes colorectal cancer is unclear. In the present study, we constructed dextran sulfate sodium (DSS)-induced colitis mouse model to uncover possible alterations in the metabolism indexes. Interestingly, after DSS diet administration, the expression of metabolism indexes and c-Myc increased. Moreover, in vitro, we treated cells with IL-6 to simulate inflammatory microenvironment and found that glucose uptake, lactate production, and lactate dehydrogenase activity increased dramatically, mirroring what were observed in vivo. In addition, the associative inhibition of STAT3 and c-Myc could significantly block the expression of metabolic enzymes. With the inhibition of STAT3/c-Myc signaling, meanwhile, the upregulation of both cell glucose uptake and lactate production by IL-6 pretreatment was reduced simultaneously. Thus, our study indicates that inflammation could induce metabolic disorder by promoting STAT3 signaling and c-Myc activity. Collectively, we find that metabolic disruptions triggered by inflammatory signaling are associated with tumorigenesis via the STAT3/c-Myc axis.

A comparative study on the physicochemical, anti-oxidative, anti-hyperglycemic and anti-lipidemic properties of amadumbe (Colocasia esculenta ) and okra (Abelmoschus esculentus ) mucilage

We investigated the physicochemical and bioactive properties of amadumbe (Colocasia esculenta) and okra (Abelmoschus esculentus) dry mucilage extracts. Protein (340 mg/g) and phenolic (40 mg/g) contents of amadumbe mucilage were 3 and 1.5 times those of okra mucilage, respectively. Amadumbe mucilage had slightly higher neutral sugar content than okra mucilage. By ¹H NMR fingerprints, rhamnopyranosyl residues were peculiar to okra mucilage. Amadumbe mucilage showed stronger radical scavenging (IC₅₀, 2.55 ± 0.30 vs. 3.01 ± 0.74 mg/mL), α‐glucosidase (IC₅₀, 1.60 ± 0.17 vs. 3.28 ± 0.5 mg/mL) and pancreatic lipase (IC₅₀, 1.63 ± 0.15 vs. 2.45 ± 0.24 mg/mL) inhibition and glucose uptake activity, perhaps due to higher phenolic content. Okra mucilage exhibited stronger intestinal glucose absorption inhibition (IC₅₀, 0.54 ± 0.24 vs. 1.82 ± 0.24 mg/mL), possibly due to higher viscosity. Data suggest that amadumbe and okra mucilage may be useful dietary supplements for managing oxidative stress, hyperglycemia, overweight and obesity. PRACTICAL APPLICATIONS: The present study reported some anti‐hyperglycemic and anti‐lipidemic potentials of dry mucilage extracted from amadumbe corm and okra pods as well as the possible modes of actions. This study is of great relevance because it can be a notable reference for further investigation on the potential use of these mucilage extracts as dietary components and/or supplements for managing hyperglycemia, overweight and obesity, thus increasing utilization and consequently improving the economic values of these traditional plants.