Mechanisms of action of the antidiabetic peptide [S4K]CPF-AM1 in db/db mice

in Journal of Molecular Endocrinology
View More View Less
  • 1 Diabetes Research Group, School of Biomedical Sciences, Ulster University, Coleraine, Northern Ireland,

Correspondence should be addressed to J M Conlon: m.conlon@ulster.ac.uk
Restricted access

The antidiabetic effects and mechanisms of action of an analogue of a frog skin host-defence peptide belonging to the caerulein-precursor fragment family, [S4K]CPF-AM1 were investigated in db/db mice with a genetically inherited form of degenerative diabetes-obesity. Twice-daily treatment with the peptide (75 nmol/kg body weight) for 28 days significantly decreased blood glucose (P < 0.01) and HbA1c (P < 0.05) and increased plasma insulin (P < 0.05) concentrations with no effect on body weight, energy intake, body composition or plasma lipid profile. Peptide administration improved insulin sensitivity and intraperitoneal glucose tolerance. Elevated biomarkers of liver and kidney function associated with the db/db phenotype were significantly lowered by [S4K]CPF-AM1 administration. Peptide treatment significantly (P < 0.05) increased pancreatic insulin content and improved the responses of isolated islets to established secretagogues. Elevated expression of genes associated with insulin signalling (Slc2a4, Insr, Irs1, Akt1, Pik3ca, Ppm1b) in the skeletal muscle of db/db mice were significantly downregulated by peptide treatment. Genes associated with insulin secretion (Abcc8, Kcnj11, Slc2a2, Cacn1c, Glp1r, Gipr) were significantly upregulated by treatment with [S4K]CPF-AM1. Studies with BRIN-BD1I clonal β-cells demonstrated that the peptide evoked membrane depolarisation, increased intracellular Ca2+ and cAMP and activated the protein kinase C pathway. The data indicate that the antidiabetic properties of [S4K]CPF-AM1 mice are mediated by direct insulinotropic action and by regulation of transcription of genes involved in both the secretion and action of insulin.

 

Society for Endocrinology

Sept 2018 onwards Past Year Past 30 Days
Abstract Views 33 33 33
Full Text Views 0 0 0
PDF Downloads 0 0 0
  • Abdel-Wahab YH, Power GJ, Ng MT, Flatt PR & Conlon JM 2008 Insulin-releasing properties of the frog skin peptide pseudin-2 and its [Lys18]-substituted analogue. Biological Chemistry 389 143148. (https://doi.org/10.1515/BC.2008.018)

    • Search Google Scholar
    • Export Citation
  • Abdel-Wahab YH, Patterson S, Flatt PR & Conlon JM 2010 Brevinin-2-related peptide and its [D4K] analogue stimulate insulin release in vitro and improve glucose tolerance in mice fed a high fat diet. Hormone and Metabolic Research 42 652656. (https://doi.org/10.1055/s-0030-1254126)

    • Search Google Scholar
    • Export Citation
  • Basu S, Yoffe P, Hills N & Lustig RH 2013 The relationship of sugar to population-level diabetes prevalence: an econometric analysis of repeated cross-sectional data. PLoS ONE 8 e57873. (https://doi.org/10.1371/journal.pone.0057873)

    • Search Google Scholar
    • Export Citation
  • Bora K, Borah M, Chutia H, Nath CK, Das D & Ruram AA 2016 Presence of concurrent derangements of liver function tests in type 2 diabetes and their relationship with glycemic status: a retrospective observational study from Meghalaya. Journal of Laboratory Physicians 8 3035. (https://doi.org/10.4103/0974-2727.176227)

    • Search Google Scholar
    • Export Citation
  • Brandt SJ, Götz A, Tschöp MH & Müller TD 2018 Gut hormone polyagonists for the treatment of type 2 diabetes. Peptides 100 190201. (https://doi.org/10.1016/j.peptides.2017.12.021)

    • Search Google Scholar
    • Export Citation
  • Butler AE, Janson J, Bonner-Weir S, Ritzel R, Rizza RA & Butler PC 2003 β-cell deficit and increased β-cell apoptosis in humans with type 2 diabetes. Diabetes 52 102110. (https://doi.org/10.2337/diabetes.52.1.102)

    • Search Google Scholar
    • Export Citation
  • Conlon JM & Mechkarska M 2014 Host-defense peptides with therapeutic potential from skin secretions of frogs from the family pipidae. Pharmaceuticals 7 5877. (https://doi.org/10.3390/ph7010058)

    • Search Google Scholar
    • Export Citation
  • Conlon JM, Al-Ghaferi N, Abraham B & Leprince J 2007 Strategies for transformation of naturally-occuring amphibian antimicrobial peptides into therapeutically valuable anti-infective agents. Methods 42 349357. (https://doi.org/10.1016/j.ymeth.2007.01.004)

    • Search Google Scholar
    • Export Citation
  • Conlon JM, Al-Ghaferi N, Ahmed E, Meetani MA, Leprince J & Nielsen PF 2010 Orthologs of magainin, PGLa, procaerulein-derived, and proxenopsin-derived peptides from skin secretions of the octoploid frog Xenopus amieti (Pipidae). Peptides 31 989994. (https://doi.org/10.1016/j.peptides.2010.03.002)

    • Search Google Scholar
    • Export Citation
  • Conlon JM, Mechkarska M, Lukic ML & Flatt PR 2014 Potential therapeutic applications of multifunctional host-defense peptides from frog skin as anti-cancer, anti-viral, immunomodulatory, and anti-diabetic agents. Peptides 57 6777. (https://doi.org/10.1016/j.peptides.2014.04.019)

    • Search Google Scholar
    • Export Citation
  • Conlon JM, Mechkarska M, Abdel-Wahab YH & Flatt PR 2018 Peptides from frog skin with potential for development into agents for Type 2 diabetes therapy. Peptides 100 275281. (https://doi.org/10.1016/j.peptides.2017.09.001)

    • Search Google Scholar
    • Export Citation
  • Conlon JM, Mechkarska M & Leprince J 2019 Peptidomic analysis in the discovery of therapeutically valuable peptides in amphibian skin secretions. Expert Review of Proteomics 16 897908. (https://doi.org/10.1080/14789450.2019.1693894)

    • Search Google Scholar
    • Export Citation
  • DeFronzo RA & Tripathy D 2009 Skeletal muscle insulin resistance is the primary defect in type 2 diabetes. Diabetes Care 32 (Supplement 2) S157S163. (https://doi.org/10.2337/dc09-S302)

    • Search Google Scholar
    • Export Citation
  • Do OH, Low JT, Gaisano HY & Thorn P 2014 The secretory deficit in islets from db/db mice is mainly due to a loss of responding beta cells. Diabetologia 57 14001409. (https://doi.org/10.1007/s00125-014-3226-8)

    • Search Google Scholar
    • Export Citation
  • Feliers D, Duraisamy S, Faulkner JL, Duch J, Lee AV, Abboud HE, Choudhury GG & Kasinath BS 2001 Activation of renal signaling pathways in db/db mice with type 2 diabetes. Kidney International 60 495504. (https://doi.org/10.1046/j.1523-1755.2001.060002495.x)

    • Search Google Scholar
    • Export Citation
  • Filippatos TD, Panagiotopoulou TV & Elisaf MS 2014 Adverse effects of GLP-1 receptor agonists. Review of Diabetic Studies 11 202230. (https://doi.org/10.1900/RDS.2014.11.202)

    • Search Google Scholar
    • Export Citation
  • Flatt PR & Bailey CJ 1981 Abnormal plasma glucose and insulin responses in heterozygous lean (ob/+) mice. Diabetologia 20 573577. (https://doi.org/10.1007/BF00252768)

    • Search Google Scholar
    • Export Citation
  • Gotoh M, Maki T, Kiyoizumi T, Satomi S & Monaco AP 1985 An improved method for isolation of mouse pancreatic islets. Transplantation 40 437438. (https://doi.org/10.1097/00007890-198510000-00018)

    • Search Google Scholar
    • Export Citation
  • Graham GV, McCloskey A, Abdel-Wahab YH, Conlon JM & Flatt PR 2020 A long-acting, dual-agonist analogue of lamprey GLP-1 shows potent insulinotropic, β-cell protective, and anorexic activities and improves glucose homeostasis in high fat-fed mice. Molecular and Cellular Endocrinology 499 110584. (https://doi.org/10.1016/j.mce.2019.110584)

    • Search Google Scholar
    • Export Citation
  • Jessen N, Buhl ES, Schmitz O & Lund S 2006 Impaired insulin action despite upregulation of proximal insulin signaling: novel insights into skeletal muscle insulin resistance in liver cirrhosis. Journal of Hepatology 45 797804. (https://doi.org/10.1016/j.jhep.2006.07.035)

    • Search Google Scholar
    • Export Citation
  • Kahn SE, Hull RL & Utzschneider KM 2006 Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature 444 840846. (https://doi.org/10.1038/nature05482)

    • Search Google Scholar
    • Export Citation
  • Katz A, Nambi SS, Mather K, Baron AD, Follmann DA, Sullivan G & Quon MJ 2000 Quantitative insulin sensitivity check index: a simple, accurate method for assessing insulin sensitivity in humans. Journal of Clinical Endocrinology and Metabolism 85 24022410. (https://doi.org/10.1210/jcem.85.7.6661)

    • Search Google Scholar
    • Export Citation
  • Martins J, Olorunju SA, Murray LM & Pillay TS 2015 Comparison of equations for the calculation of LDL-cholesterol in hospitalized patients. Clinica Chimica Acta: International Journal of Clinical Chemistry 444 137142. (https://doi.org/10.1016/j.cca.2015.01.037)

    • Search Google Scholar
    • Export Citation
  • McClenaghan NH, Barnett CR, Ah-Sing E, Abdel-Wahab YHA, O’Harte FP, Yoon TW, Swanston-Flatt SK & Flatt PR 1996 Characterization of a novel glucose-responsive insulin-secreting cell line, BRIN-BD11, produced by electrofusion. Diabetes 45 11321140. (https://doi.org/10.2337/diab.45.8.1132)

    • Search Google Scholar
    • Export Citation
  • McLean DT, McCrudden MT, Linden GJ, Irwin CR, Conlon JM & Lundy FT 2014 Antimicrobial and immunomodulatory properties of PGLa-AM1, CPF-AM1, and magainin-AM1: potent activity against oral pathogens. Regulatory Peptides 194 195 6368. (https://doi.org/10.1016/j.regpep.2014.11.002)

    • Search Google Scholar
    • Export Citation
  • O’Harte FPM, Parthsarathy V, Hogg C & Flatt PR 2018 Long-term treatment with acylated analogues of apelin-13 amide ameliorates diabetes and improves lipid profile of high-fat fed mice. PLoS ONE 13 e0202350. (https://doi.org/10.1371/journal.pone.0202350)

    • Search Google Scholar
    • Export Citation
  • Ojo OO, Abdel-Wahab YH, Flatt PR & Conlon JM 2013a Insulinotropic actions of the frog skin host-defense peptide alyteserin-2a: a structure-activity study. Chemical Biology and Drug Design 82 196204. (https://doi.org/10.1111/cbdd.12151)

    • Search Google Scholar
    • Export Citation
  • Ojo OO, Conlon JM, Flatt PR & Abdel-Wahab YH 2013b Frog skin peptides (tigerinin-1R, magainin-AM1, -AM2, CPF-AM1, and PGla-AM1) stimulate secretion of glucagon-like peptide 1 (GLP-1) by GLUTag cells. Biochemical and Biophysical Research Communications 431 1418. (https://doi.org/10.1016/j.bbrc.2012.12.116)

    • Search Google Scholar
    • Export Citation
  • Ojo OO, Srinivasan DK, Owolabi BO, McGahon MK, Moffett RC, Curtis TM, Conlon JM, Flatt PR & Abdel-Wahab YH 2016 Molecular mechanisms mediating the beneficial metabolic effects of [Arg4] tigerinin-1R in mice with diet-induced obesity and insulin resistance. Biological Chemistry 397 753764. (https://doi.org/10.1515/hsz-2016-0120)

    • Search Google Scholar
    • Export Citation
  • Owolabi BO, Ojo OO, Srinivasan DK, Conlon JM, Flatt PR & Abdel-Wahab YH 2016a Glucoregulatory, endocrine and morphological effects of [P5K]hymenochirin-1B in mice with diet-induced glucose intolerance and insulin resistance. Naunyn-Schmiedeberg’s Archives of Pharmacology 389 769781. (https://doi.org/10.1007/s00210-016-1243-5)

    • Search Google Scholar
    • Export Citation
  • Owolabi BO, Ojo OO, Srinivasan DK, Conlon JM, Flatt PR & Abdel-Wahab YH 2016b In vitro and in vivo insulinotropic properties of the multifunctional frog skin peptide hymenochirin-1B: a structure–activity study. Amino Acids 48 535547. (https://doi.org/10.1007/s00726-015-2107-x)

    • Search Google Scholar
    • Export Citation
  • Owolabi BO, Musale V, Ojo OO, Moffett RC, McGahon MK, Curtis TM, Conlon JM, Flatt PR & Abdel-Wahab YHA 2017 Actions of PGLa-AM1 and its [A14K] and [A20K] analogues and their therapeutic potential as anti-diabetic agents. Biochimie 138 112. (https://doi.org/10.1016/j.biochi.2017.04.004)

    • Search Google Scholar
    • Export Citation
  • Pantic JM, Jovanovic IP, Radosavljevic GD, Arsenijevic NN, Conlon JM & Lukic ML 2017 The potential of frog skin-derived peptides for development into therapeutically-valuable immunomodulatory agents. Molecules 22 E2071. (https://doi.org/10.3390/molecules22122071)

    • Search Google Scholar
    • Export Citation
  • Park CW, Kim HW, Ko SH, Lim JH, Ryu GR, Chung HW, Han SW, Shin SJ, Bang BK & Breyer MD 2007 Long-term treatment of glucagon-like peptide-1 analog exendin-4 ameliorates diabetic nephropathy through improving metabolic anomalies in db/db mice. Journal of the American Society of Nephrology 18 12271238. (https://doi.org/10.1681/ASN.2006070778)

    • Search Google Scholar
    • Export Citation
  • Parkes DG, Mace KF & Trautmann ME 2013 Discovery and development of exenatide: the first antidiabetic agent to leverage the multiple benefits of the incretin hormone, GLP-1. Expert Opinion on Drug Discovery 8 219244. (https://doi.org/10.1517/17460441.2013.741580)

    • Search Google Scholar
    • Export Citation
  • Parthsarathy V, Irwin N, Hasib A, Martin CM, McClean S, Bhat VK, Ng MT, Flatt PR & Gault VA 2016 A novel chemically modified analogue of xenin-25 exhibits improved glucose-lowering and insulin-releasing properties. Biochimica et Biophysica Acta 1860 757764. (https://doi.org/10.1016/j.bbagen.2016.01.015)

    • Search Google Scholar
    • Export Citation
  • Poitout V 2013 Lipotoxicity impairs incretin signalling. Diabetologia 56 231233. (https://doi.org/10.1007/s00125-012-2788-6)

  • Shimoda M, Kanda Y, Hamamoto S, Tawaramoto K, Hashiramoto M, Matsuki M & Kaku K 2011 The human glucagon-like peptide-1 analogue liraglutide preserves pancreatic beta cells via regulation of cell kinetics and suppression of oxidative and endoplasmic reticulum stress in a mouse model of diabetes. Diabetologia 54 10981108. (https://doi.org/10.1007/s00125-011-2069-9)

    • Search Google Scholar
    • Export Citation
  • Son DJ, Hwang SY, Kim MH, Park UK & Kim BS 2015 Anti-diabetic and hepato-renal protective effects of Ziyuglycoside II methyl ester in type 2 diabetic mice. Nutrients 7 54695483. (https://doi.org/10.3390/nu7075232)

    • Search Google Scholar
    • Export Citation
  • Srinivasan D, Mechkarska M, Abdel-Wahab YH, Flatt PR & Conlon JM 2013 Caerulein precursor fragment (CPF) peptides from the skin secretions of Xenopus laevis and Silurana epitropicalis are potent insulin-releasing agents. Biochimie 95 429435. (https://doi.org/10.1016/j.biochi.2012.10.026)

    • Search Google Scholar
    • Export Citation
  • Srinivasan D, Ojo OO, Abdel-Wahab YH, Flatt PR, Guilhaudis L & Conlon JM 2014 Insulin-releasing and cytotoxic properties of the frog skin peptide, tigerinin-1R: a structure-activity study. Peptides 55 2331. (https://doi.org/10.1016/j.peptides.2014.02.002)

    • Search Google Scholar
    • Export Citation
  • Srinivasan D, Ojo OO, Owolabi BO, Conlon JM, Flatt PR & Abdel-Wahab YHA 2015 The frog skin host-defense peptide CPF-SE1 improves glucose tolerance, insulin sensitivity and islet function and decreases plasma lipids in high-fat fed mice. European Journal of Pharmacology 764 3847. (https://doi.org/10.1016/j.ejphar.2015.06.042)

    • Search Google Scholar
    • Export Citation
  • Steinberg WM, Buse JB, Ghorbani MLM, Ørsted DD, Nauck MALEADER Steering Committee & LEADER Trial Investigators 2017 Amylase, lipase, and acute pancreatitis in people with type 2 diabetes treated with liraglutide: results from the LEADER randomized trial. Diabetes Care 40 966972. (https://doi.org/10.2337/dc16-2747)

    • Search Google Scholar
    • Export Citation
  • Thorens B, Porret A, Buhler L, Deng SP, Morel P & Widmann C 1993 Cloning and functional expression of the human islet GLP-1 receptor. Demonstration that exendin-4 is an agonist and exendin-(9–39) an antagonist of the receptor. Diabetes 42 16781682. (https://doi.org/10.2337/diab.42.11.1678)

    • Search Google Scholar
    • Export Citation
  • UK Prospective Diabetes Study (UKPDS) Group 1998 Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). Lancet 352 837853. (https://doi.org/10.1016/S0140-6736(9807019-6)

    • Search Google Scholar
    • Export Citation
  • Vasu S, Moffett RC, Thorens B & Flatt PR 2014 Role of endogenous GLP-1 and GIP in beta cell compensatory responses to insulin resistance and cellular stress. PLoS ONE 9 e101005. (https://doi.org/10.1371/journal.pone.0101005)

    • Search Google Scholar
    • Export Citation
  • Vasu S, McGahon MK, Moffett RC, Curtis TM, Conlon JM, Abdel-Wahab YH & Flatt PR 2017 Esculentin-2CHa(1–30) and its analogues: stability and mechanisms of insulinotropic action. Journal of Endocrinology 232 423435. (https://doi.org/10.1530/JOE-16-0453)

    • Search Google Scholar
    • Export Citation
  • Wang Q & Brubaker PL 2002 Glucagon-like peptide-1 treatment delays the onset of diabetes in 8 week-old db/db mice. Diabetologia 45 12631273. (https://doi.org/10.1007/s00125-002-0828-3)

    • Search Google Scholar
    • Export Citation
  • Xu X & Lai R 2015 The chemistry and biological activities of peptides from amphibian skin secretions. Chemical Reviews 115 17601846. (https://doi.org/10.1021/cr4006704)

    • Search Google Scholar
    • Export Citation
  • Yang Q, Xu Y, Xie P, Cheng H, Song Q, Su T, Yuan S & Liu Q 2015 Retinal neurodegeneration in db/db mice at the early period of diabetes. Journal of Ophthalmology 2015 757412. (https://doi.org/10.1155/2015/757412)

    • Search Google Scholar
    • Export Citation