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  1. Article ; Online: Preanalytical impact on the accuracy of measurements of glucagon, GLP-1 and GIP in clinical trials.

    Rasmussen, Christine / Richter, Michael M / Jensen, Nicole J / Heinz, Niklas / Hartmann, Bolette / Holst, Jens J / Kjeldsen, Sasha A S / Wewer Albrechtsen, Nicolai J

    Scandinavian journal of clinical and laboratory investigation

    2024  Volume 83, Issue 8, Page(s) 591–598

    Abstract: Background: Plasma concentrations of glucagon, GLP-1 and GIP are reported in numerous clinical trials as outcome measures but preanalytical guidelines are lacking. We addressed the impact of commonly used blood containers in metabolic research on ... ...

    Abstract Background: Plasma concentrations of glucagon, GLP-1 and GIP are reported in numerous clinical trials as outcome measures but preanalytical guidelines are lacking. We addressed the impact of commonly used blood containers in metabolic research on measurements of glucagon, GLP-1 and GIP in humans.
    Methods: Seventeen overweight individuals were subjected to an overnight fast followed by an intravenous infusion of amino acids to stimulate hormonal secretion. Blood was sampled into five containers: EDTA-coated tubes supplemented with DMSO (control), a neprilysin inhibitor, aprotinin (a kallikrein inhibitor) or a DPP-4 inhibitor, and P800 tubes. Plasma was kept on ice before and after centrifugation and stored at -80 Celsius until batch analysis using validated sandwich ELISAs or radioimmunoassays (RIA).
    Results: Measures of fasting plasma glucagon did not depend on sampling containers, whether measured by ELISA or RIA. Amino acid-induced hyperglucagonemia was numerically higher when blood was collected into P800 tubes or tubes with aprotinin. The use of p800 tubes resulted in higher concentrations of GLP-1 by RIA compared to control tubes but not for measurements with sandwich ELISA. Plasma concentrations of GIP measured by ELISA were higher in control tubes and negatively affected by P800 and the addition of aprotinin.
    Conclusions: The choice of blood containers impacts on measurements of plasma concentrations of glucagon, GLP-1 and GIP, and based on this study, we recommend using EDTA-coated tubes without protease inhibitors or P800 tubes for measurements of glucagon, GLP-1 and GIP in clinical trials.
    MeSH term(s) Humans ; Glucagon/metabolism ; Glucagon-Like Peptide 1 ; Aprotinin ; Edetic Acid ; Gastric Inhibitory Polypeptide/metabolism ; Blood Glucose/analysis ; Insulin ; Peptide Fragments
    Chemical Substances Glucagon (9007-92-5) ; Glucagon-Like Peptide 1 (89750-14-1) ; Aprotinin (9087-70-1) ; Edetic Acid (9G34HU7RV0) ; Gastric Inhibitory Polypeptide (59392-49-3) ; Blood Glucose ; Insulin ; Peptide Fragments
    Language English
    Publishing date 2024-01-24
    Publishing country England
    Document type Journal Article
    ZDB-ID 3150-1
    ISSN 1502-7686 ; 0036-5513
    ISSN (online) 1502-7686
    ISSN 0036-5513
    DOI 10.1080/00365513.2023.2294470
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  2. Article ; Online: Glucagon does not directly stimulate pituitary secretion of ACTH, GH or copeptin.

    Stangerup, Ida / Kjeldsen, Sasha A S / Richter, Michael M / Jensen, Nicole J / Rungby, Jørgen / Haugaard, Steen Bendix / Georg, Birgitte / Hannibal, Jens / Møllgård, Kjeld / Wewer Albrechtsen, Nicolai J / Bjørnbak Holst, Camilla

    Peptides

    2024  Volume 176, Page(s) 171213

    Abstract: Glucagon is best known for its contribution to glucose regulation through activation of the glucagon receptor (GCGR), primarily located in the liver. However, glucagon's impact on other organs may also contribute to its potent effects in health and ... ...

    Abstract Glucagon is best known for its contribution to glucose regulation through activation of the glucagon receptor (GCGR), primarily located in the liver. However, glucagon's impact on other organs may also contribute to its potent effects in health and disease. Given that glucagon-based medicine is entering the arena of anti-obesity drugs, elucidating extrahepatic actions of glucagon are of increased importance. It has been reported that glucagon may stimulate secretion of arginine-vasopressin (AVP)/copeptin, growth hormone (GH) and adrenocorticotrophic hormone (ACTH) from the pituitary gland. Nevertheless, the mechanisms and whether GCGR is present in human pituitary are unknown. In this study we found that intravenous administration of 0.2 mg glucagon to 14 healthy subjects was not associated with increases in plasma concentrations of copeptin, GH, ACTH or cortisol over a 120-min period. GCGR immunoreactivity was present in the anterior pituitary but not in cells containing GH or ACTH. Collectively, glucagon may not directly stimulate secretion of GH, ACTH or AVP/copeptin in humans but may instead be involved in yet unidentified pituitary functions.
    MeSH term(s) Humans ; Glycopeptides/metabolism ; Glucagon/metabolism ; Glucagon/blood ; Adrenocorticotropic Hormone/blood ; Adrenocorticotropic Hormone/metabolism ; Male ; Adult ; Female ; Pituitary Gland/metabolism ; Pituitary Gland/drug effects ; Hydrocortisone/blood ; Receptors, Glucagon/metabolism ; Human Growth Hormone/metabolism ; Growth Hormone/metabolism ; Growth Hormone/blood ; Middle Aged
    Chemical Substances copeptins ; Glycopeptides ; Glucagon (9007-92-5) ; Adrenocorticotropic Hormone (9002-60-2) ; Hydrocortisone (WI4X0X7BPJ) ; Receptors, Glucagon ; Human Growth Hormone (12629-01-5) ; Growth Hormone (9002-72-6)
    Language English
    Publishing date 2024-04-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 769028-9
    ISSN 1873-5169 ; 0196-9781
    ISSN (online) 1873-5169
    ISSN 0196-9781
    DOI 10.1016/j.peptides.2024.171213
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  3. Article ; Online: Glucagon augments the secretion of FGF21 and GDF15 in MASLD by indirect mechanisms.

    Richter, Michael M / Kemp, Ida M / Heebøll, Sara / Winther-Sørensen, Marie / Kjeldsen, Sasha A S / Jensen, Nicole J / Nybing, Janus D / Linden, Frederik H / Høgh-Schmidt, Erik / Boesen, Mikael P / Madsbad, Sten / Schiødt, Frank Vinholt / Nørgaard, Kirsten / Schmidt, Signe / Gluud, Lise Lotte / Haugaard, Steen B / Holst, Jens J / Nielsen, Søren / Rungby, Jørgen /
    Wewer Albrechtsen, Nicolai J

    Metabolism: clinical and experimental

    2024  , Page(s) 155915

    Abstract: Introduction: Glucagon receptor agonism is currently explored for the treatment of obesity and metabolic dysfunction-associated steatotic liver disease (MASLD). The metabolic effects of glucagon receptor agonism may in part be mediated by increases in ... ...

    Abstract Introduction: Glucagon receptor agonism is currently explored for the treatment of obesity and metabolic dysfunction-associated steatotic liver disease (MASLD). The metabolic effects of glucagon receptor agonism may in part be mediated by increases in circulating levels of Fibroblast Growth Factor 21 (FGF21) and Growth Differentiation Factor 15 (GDF15). The effect of glucagon agonism on FGF21 and GDF15 levels remains uncertain, especially in the context of elevated insulin levels commonly observed in metabolic diseases.
    Methods: We investigated the effect of a single bolus of glucagon and a continuous infusion of glucagon on plasma concentrations of FGF21 and GDF15 in conditions of endogenous low or high insulin levels. The studies included individuals with overweight with and without MASLD, healthy controls (CON) and individuals with type 1 diabetes (T1D). The direct effect of glucagon on FGF21 and GDF15 was evaluated using our in-house developed isolated perfused mouse liver model.
    Results: FGF21 and GDF15 correlated with plasma levels of insulin, but not glucagon, and their secretion were highly increased in MASLD compared with CON and T1D. Furthermore, FGF21 levels in individuals with overweight with or without MASLD did not increase after glucagon stimulation when insulin levels were kept constant. FGF21 and GDF15 levels were unaffected by direct stimulation with glucagon in the isolated perfused mouse liver.
    Conclusion: The glucagon-induced secretion of FGF21 and GDF15 are augmented in MASLD and may depend on insulin. Thus, glucagon receptor agonism may augment its metabolic benefits in patients with MASLD through enhanced secretion of FGF21 and GDF15.
    Language English
    Publishing date 2024-04-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 80230-x
    ISSN 1532-8600 ; 0026-0495
    ISSN (online) 1532-8600
    ISSN 0026-0495
    DOI 10.1016/j.metabol.2024.155915
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  4. Article ; Online: Development of a glucagon sensitivity test in humans: Pilot data and the GLUSENTIC study protocol

    Kjeldsen, Sasha A.S. / Richter, Michael M. / Jensen, Nicole J. / Nilsson, Malin S.D. / Heinz, Niklas / Nybing, Janus D. / Linden, Frederik H. / Høgh-Schmidt, Erik / Boesen, Mikael P. / Madsbad, Sten / Vilstrup, Hendrik / Schiødt, Frank Vinholt / Møller, Andreas / Nørgaard, Kirsten / Schmidt, Signe / Rashu, Elias B. / Gluud, Lise L. / Haugaard, Steen B. / Holst, Jens J. /
    Rungby, Jørgen / Wewer Albrechtsen, Nicolai J.

    Peptides. 2023 Mar., v. 161 p.170938-

    2023  

    Abstract: A physiological feedback system exists between hepatocytes and the alpha cells, termed the liver-alpha cell axis and refers to the relationship between amino acid-stimulated glucagon secretion and glucagon-stimulated amino acid catabolism. Several ... ...

    Abstract A physiological feedback system exists between hepatocytes and the alpha cells, termed the liver-alpha cell axis and refers to the relationship between amino acid-stimulated glucagon secretion and glucagon-stimulated amino acid catabolism. Several reports indicate that non-alcoholic fatty liver disease (NAFLD) disrupts the liver-alpha cell axis, because of impaired glucagon receptor signaling (glucagon resistance). However, no experimental test exists to assess glucagon resistance in humans. The objective was to develop an experimental test to determine glucagon sensitivity with respect to amino acid and glucose metabolism in humans. The proposed glucagon sensitivity test (comprising two elements: 1) i.v. injection of 0.2 mg glucagon and 2) infusion of mixed amino acids 331 mg/hour/kg) is based on nine pilot studies which are presented. Calculation of a proposed glucagon sensitivity index with respect to amino acid catabolism is also described. Secondly, we describe a complete study protocol (GLUSENTIC) according to which the glucagon sensitivity test will be applied in a cross-sectional study currently taking place. 65 participants including 20 individuals with a BMI 18.6–25 kg/m², 30 individuals with a BMI ≥ 25–40 kg/m², and 15 individuals with type 1 diabetes with a BMI between 18.6 and 40 kg/m² will be included. Participants will be grouped according to their degree of hepatic steatosis measured by whole-liver magnetic resonance imaging (MRI). The primary outcome measure will be differences in the glucagon sensitivity index between individuals with and without hepatic steatosis. Developing a glucagon sensitivity test and index may provide insight into the physiological and pathophysiological mechanism of glucagon action and glucagon-based therapies.
    Keywords amino acid metabolism ; amino acids ; cross-sectional studies ; fatty liver ; glucagon ; glucagon receptors ; glucose ; hepatocytes ; insulin-dependent diabetes mellitus ; magnetism ; secretion ; NAFLD ; Type 1 diabetes ; Metabolism ; The liver-alpha cell axis ; Study protocol
    Language English
    Dates of publication 2023-03
    Publishing place Elsevier Inc.
    Document type Article ; Online
    Note Use and reproduction
    ZDB-ID 769028-9
    ISSN 1873-5169 ; 0196-9781
    ISSN (online) 1873-5169
    ISSN 0196-9781
    DOI 10.1016/j.peptides.2022.170938
    Database NAL-Catalogue (AGRICOLA)

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  5. Article ; Online: Neprilysin activity is increased in metabolic dysfunction-associated steatotic liver disease and normalizes after bariatric surgery or GLP-1 therapy.

    Kjeldsen, Sasha A S / Gluud, Lise L / Werge, Mikkel P / Pedersen, Julie S / Bendtsen, Flemming / Alexiadou, Kleopatra / Tan, Tricia / Torekov, Signe S / Iepsen, Eva W / Jensen, Nicole J / Richter, Michael M / Goetze, Jens P / Rungby, Jørgen / Hartmann, Bolette / Holst, Jens J / Holst, Birgitte / Holt, Joachim / Gustafsson, Finn / Madsbad, Sten /
    Svane, Maria S / Bojsen-Møller, Kirstine N / Wewer Albrechtsen, Nicolai J

    iScience

    2023  Volume 26, Issue 11, Page(s) 108190

    Abstract: Inhibitors of neprilysin improve glycemia in patients with heart failure and type 2 diabetes (T2D). The effect of weight loss by diet, surgery, or pharmacotherapy on neprilysin activity (NEPa) is unknown. We investigated circulating NEPa and neprilysin ... ...

    Abstract Inhibitors of neprilysin improve glycemia in patients with heart failure and type 2 diabetes (T2D). The effect of weight loss by diet, surgery, or pharmacotherapy on neprilysin activity (NEPa) is unknown. We investigated circulating NEPa and neprilysin protein concentrations in obesity, T2D, metabolic dysfunction-associated steatotic liver disease (MASLD), and following bariatric surgery, or GLP-1-receptor-agonist therapy. NEPa, but not neprilysin protein, was enhanced in obesity, T2D, and MASLD. Notably, MASLD associated with NEPa independently of BMI and HbA1c. NEPa decreased after bariatric surgery with a concurrent increase in OGTT-stimulated GLP-1. Diet-induced weight loss did not affect NEPa, but individuals randomized to 52-week weight maintenance with liraglutide (1.2 mg/day) decreased NEPa, consistent with another study following 6-week liraglutide (3 mg/day). A 90-min GLP-1 infusion did not alter NEPa. Thus, MASLD may drive exaggerated NEPa, and lowered NEPa following bariatric surgery or liraglutide therapy may contribute to the reported improved cardiometabolic effects.
    Language English
    Publishing date 2023-10-12
    Publishing country United States
    Document type Journal Article
    ISSN 2589-0042
    ISSN (online) 2589-0042
    DOI 10.1016/j.isci.2023.108190
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  6. Article ; Online: On measurements of glucagon secretion in healthy, obese, and Roux-en-Y gastric bypass operated individuals using sandwich ELISA.

    Wewer Albrechtsen, Nicolai J / Kjeldsen, Sasha A S / Jensen, Nicole J / Rungby, Jørgen / Veedfald, Simon / Bojsen-Møller, Kirstine N / Dirksen, Carsten / Jensen, Christian Z / Martinussen, Christoffer / Madsbad, Sten / Holst, Jens J

    Scandinavian journal of clinical and laboratory investigation

    2021  Volume 82, Issue 1, Page(s) 75–83

    Abstract: Glucagon is a key regulator of metabolism and is used in the diagnostic of neuroendocrine tumors. Accurate measurement of glucagon requires both extreme sensitivity and specificity since several peptides are derived from the same proglucagon precursor ... ...

    Abstract Glucagon is a key regulator of metabolism and is used in the diagnostic of neuroendocrine tumors. Accurate measurement of glucagon requires both extreme sensitivity and specificity since several peptides are derived from the same proglucagon precursor encoding part of the glucagon sequence and given that glucagon circulates in picomolar concentrations. A sandwich ELISA was recently developed and extensively evaluated; however, this method may not be accurate when measuring glucagon in patients with an enhanced production of proglucagon-derived peptides as seen after Roux-en-Y gastric bypass (RYGB). To overcome this, a modified version of the ELISA was developed. In this study, we evaluate an unmodified and a modified version of the ELISA in healthy individuals, individuals with obesity, and finally in two cohorts of patients following RYGB surgery using different nutrient stimuli to assess glucagon dynamics. Finally,
    MeSH term(s) Blood Glucose/analysis ; Enzyme-Linked Immunosorbent Assay ; Gastric Bypass/methods ; Glucagon/metabolism ; Humans ; Obesity/surgery ; Proglucagon
    Chemical Substances Blood Glucose ; Proglucagon (55963-74-1) ; Glucagon (9007-92-5)
    Language English
    Publishing date 2021-12-22
    Publishing country England
    Document type Journal Article
    ZDB-ID 3150-1
    ISSN 1502-7686 ; 0036-5513
    ISSN (online) 1502-7686
    ISSN 0036-5513
    DOI 10.1080/00365513.2021.2016943
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Development of a glucagon sensitivity test in humans: Pilot data and the GLUSENTIC study protocol.

    Kjeldsen, Sasha A S / Richter, Michael M / Jensen, Nicole J / Nilsson, Malin S D / Heinz, Niklas / Nybing, Janus D / Linden, Frederik H / Høgh-Schmidt, Erik / Boesen, Mikael P / Madsbad, Sten / Vilstrup, Hendrik / Schiødt, Frank Vinholt / Møller, Andreas / Nørgaard, Kirsten / Schmidt, Signe / Rashu, Elias B / Gluud, Lise L / Haugaard, Steen B / Holst, Jens J /
    Rungby, Jørgen / Wewer Albrechtsen, Nicolai J

    Peptides

    2022  Volume 161, Page(s) 170938

    Abstract: A physiological feedback system exists between hepatocytes and the alpha cells, termed the liver-alpha cell axis and refers to the relationship between amino acid-stimulated glucagon secretion and glucagon-stimulated amino acid catabolism. Several ... ...

    Abstract A physiological feedback system exists between hepatocytes and the alpha cells, termed the liver-alpha cell axis and refers to the relationship between amino acid-stimulated glucagon secretion and glucagon-stimulated amino acid catabolism. Several reports indicate that non-alcoholic fatty liver disease (NAFLD) disrupts the liver-alpha cell axis, because of impaired glucagon receptor signaling (glucagon resistance). However, no experimental test exists to assess glucagon resistance in humans. The objective was to develop an experimental test to determine glucagon sensitivity with respect to amino acid and glucose metabolism in humans. The proposed glucagon sensitivity test (comprising two elements: 1) i.v. injection of 0.2 mg glucagon and 2) infusion of mixed amino acids 331 mg/hour/kg) is based on nine pilot studies which are presented. Calculation of a proposed glucagon sensitivity index with respect to amino acid catabolism is also described. Secondly, we describe a complete study protocol (GLUSENTIC) according to which the glucagon sensitivity test will be applied in a cross-sectional study currently taking place. 65 participants including 20 individuals with a BMI 18.6-25 kg/m
    MeSH term(s) Humans ; Glucagon/metabolism ; Cross-Sectional Studies ; Liver/metabolism ; Non-alcoholic Fatty Liver Disease/metabolism ; Amino Acids
    Chemical Substances Glucagon (9007-92-5) ; Amino Acids
    Language English
    Publishing date 2022-12-31
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 769028-9
    ISSN 1873-5169 ; 0196-9781
    ISSN (online) 1873-5169
    ISSN 0196-9781
    DOI 10.1016/j.peptides.2022.170938
    Database MEDical Literature Analysis and Retrieval System OnLINE

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