GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients

Abstract Metformin (N,N-dimethylbiguanide), an inhibitor of gluconeogenesis and insulin sensitizer, is widely used for the treatment of type 2 diabetes. In some patients with renal insufficiency, metformin can accumulate and cause lactic acidosis, known as metformin-associated lactic acidosis (MALA,...

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Main Authors: Svetlana Baskal, Rene A. Posma, Alexander Bollenbach, Willem Dieperink, Stephan J. L. Bakker, Maarten W. Nijsten, Daan J. Touw, Dimitrios Tsikas
Format: Article
Language:English
Published: Springer 2024-03-01
Series:Amino Acids
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Online Access:https://doi.org/10.1007/s00726-024-03383-9
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author Svetlana Baskal
Rene A. Posma
Alexander Bollenbach
Willem Dieperink
Stephan J. L. Bakker
Maarten W. Nijsten
Daan J. Touw
Dimitrios Tsikas
author_facet Svetlana Baskal
Rene A. Posma
Alexander Bollenbach
Willem Dieperink
Stephan J. L. Bakker
Maarten W. Nijsten
Daan J. Touw
Dimitrios Tsikas
author_sort Svetlana Baskal
collection DOAJ
description Abstract Metformin (N,N-dimethylbiguanide), an inhibitor of gluconeogenesis and insulin sensitizer, is widely used for the treatment of type 2 diabetes. In some patients with renal insufficiency, metformin can accumulate and cause lactic acidosis, known as metformin-associated lactic acidosis (MALA, defined as lactate ≥ 5 mM, pH < 7.35, and metformin concentration > 38.7 µM). Here, we report on the post-translational modification (PTM) of proline (Pro) to 4-hydroxyproline (OH-Pro) in metformin-associated lactic acidosis and in metformin-treated patients with Becker muscular dystrophy (BMD). Pro and OH-Pro were measured simultaneously by gas chromatography–mass spectrometry before, during, and after renal replacement therapy in a patient admitted to the intensive care unit (ICU) because of MALA. At admission to the ICU, plasma metformin concentration was 175 µM, with a corresponding lactate concentration of 20 mM and a blood pH of 7.1. Throughout ICU admission, the Pro concentration was lower compared to healthy controls. Renal excretion of OH-Pro was initially high and decreased over time. Moreover, during the first 12 h of ICU admission, OH-Pro seems to be renally secreted while thereafter, it was reabsorbed. Our results suggest that MALA is associated with hyper-hydroxyprolinuria due to elevated PTM of Pro to OH-Pro by prolyl-hydroxylase and/or inhibition of OH-Pro metabolism in the kidneys. In BMD patients, metformin, at the therapeutic dose of 3 × 500 mg per day for 6 weeks, increased the urinary excretion of OH-Pro suggesting elevation of Pro hydroxylation to OH-Pro. Our study suggests that metformin induces specifically the expression/activity of prolyl-hydroxylase in metformin intoxication and BMD.
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spelling doaj-art-88c742db65bf41abae3835a9c3d64df42025-08-20T01:57:16ZengSpringerAmino Acids1438-21992024-03-0156111510.1007/s00726-024-03383-9GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patientsSvetlana Baskal0Rene A. Posma1Alexander Bollenbach2Willem Dieperink3Stephan J. L. Bakker4Maarten W. Nijsten5Daan J. Touw6Dimitrios Tsikas7Institute of Toxicology, Core Unit Proteomics, Hannover Medical SchoolDepartment of Critical Care, University of Groningen, University Medical Center GroningenInstitute of Toxicology, Core Unit Proteomics, Hannover Medical SchoolDepartment of Critical Care, University of Groningen, University Medical Center GroningenDivision of Nephrology, Department of Internal Medicine, University of Groningen, University Medical Center GroningenDepartment of Critical Care, University of Groningen, University Medical Center GroningenDepartment of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center GroningenInstitute of Toxicology, Core Unit Proteomics, Hannover Medical SchoolAbstract Metformin (N,N-dimethylbiguanide), an inhibitor of gluconeogenesis and insulin sensitizer, is widely used for the treatment of type 2 diabetes. In some patients with renal insufficiency, metformin can accumulate and cause lactic acidosis, known as metformin-associated lactic acidosis (MALA, defined as lactate ≥ 5 mM, pH < 7.35, and metformin concentration > 38.7 µM). Here, we report on the post-translational modification (PTM) of proline (Pro) to 4-hydroxyproline (OH-Pro) in metformin-associated lactic acidosis and in metformin-treated patients with Becker muscular dystrophy (BMD). Pro and OH-Pro were measured simultaneously by gas chromatography–mass spectrometry before, during, and after renal replacement therapy in a patient admitted to the intensive care unit (ICU) because of MALA. At admission to the ICU, plasma metformin concentration was 175 µM, with a corresponding lactate concentration of 20 mM and a blood pH of 7.1. Throughout ICU admission, the Pro concentration was lower compared to healthy controls. Renal excretion of OH-Pro was initially high and decreased over time. Moreover, during the first 12 h of ICU admission, OH-Pro seems to be renally secreted while thereafter, it was reabsorbed. Our results suggest that MALA is associated with hyper-hydroxyprolinuria due to elevated PTM of Pro to OH-Pro by prolyl-hydroxylase and/or inhibition of OH-Pro metabolism in the kidneys. In BMD patients, metformin, at the therapeutic dose of 3 × 500 mg per day for 6 weeks, increased the urinary excretion of OH-Pro suggesting elevation of Pro hydroxylation to OH-Pro. Our study suggests that metformin induces specifically the expression/activity of prolyl-hydroxylase in metformin intoxication and BMD.https://doi.org/10.1007/s00726-024-03383-9Amino acidsBMDDialysisGC–MS4-HydroxyprolineIntoxication
spellingShingle Svetlana Baskal
Rene A. Posma
Alexander Bollenbach
Willem Dieperink
Stephan J. L. Bakker
Maarten W. Nijsten
Daan J. Touw
Dimitrios Tsikas
GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
Amino Acids
Amino acids
BMD
Dialysis
GC–MS
4-Hydroxyproline
Intoxication
title GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
title_full GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
title_fullStr GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
title_full_unstemmed GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
title_short GC–MS analysis of 4-hydroxyproline: elevated proline hydroxylation in metformin-associated lactic acidosis and metformin-treated Becker muscular dystrophy patients
title_sort gc ms analysis of 4 hydroxyproline elevated proline hydroxylation in metformin associated lactic acidosis and metformin treated becker muscular dystrophy patients
topic Amino acids
BMD
Dialysis
GC–MS
4-Hydroxyproline
Intoxication
url https://doi.org/10.1007/s00726-024-03383-9
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