Comparative Clinical Pathology

, Volume 21, Issue 6, pp 1493–1499 | Cite as

Association of the Leu72Met polymorphism of the ghrelin gene and ghrelin level with type 2 diabetes mellitus and obesity

  • Nadida A. Gohar
  • Dina F. Elgayar
  • Ayat I. Hassan
  • Samar H. Aboulsoud
  • Mona A. Hegazy
Original Article


The aim of the study is to resolve the debated association between ghrelin level and polymorphism of ghrelin gene (Leu72Met) with obesity and type 2 diabetes mellitus. The present study was conducted on 30 diabetic middle-aged patients with normal body mass index (BMI) and 40 age-matched non-diabetic participants (20 with normal BMI and 20 obese). All participants were subjected to full medical history and examination, anthropometric measurements, and laboratory investigations which include complete lipid profile, fasting plasma glucose, glycated hemoglobin, fasting serum insulin and HOMA-IR was calculated, fasting plasma ghrelin level was determined using ELISA method, and the Leu72Met polymorphism of the ghrelin gene was screened using PCR-RFLP. The median level of fasting plasma ghrelin was higher in the diabetic group than those of the non-diabetic group (P = 0.157). However, lower median fasting plasma ghrelin level was reported in the obese group compared to those with normal BMI, yet not statistically significant (P = 0.289). In the present study, Leu72Met polymorphism was present in 13% of the diabetic subjects, 20% of the non-diabetic subjects with normal BMI, and in 30% of the obese non-diabetic participants. No statistically significant association was found between the ghrelin level and the Leu72Met polymorphism in preproghrelin gene with either obesity or type 2 diabetes mellitus.


Ghrelin Genetic polymorphism Diabetes Obesity Polymerase chain reaction (PCR) 


Conflicts of interest



  1. American Diabetes Association (2010) Diagnosis and classification of diabetes mellitus. Diabetes Care 33:S62–S66CrossRefGoogle Scholar
  2. Barazzoni R, Zanetti M, Ferreiral (2007) Relationships between desacylated and acylated ghrelin and insulin sensitivity in metabolic syndrome. J Clin Endocrinol Metab 92:3935–3940PubMedCrossRefGoogle Scholar
  3. Berthold HK, Giannakidou E, Krone W, Mantzoros CS, Gouni-Berthold I (2009) The Leu72Met polymorphism of the ghrelin gene is associated with a decreased risk for type 2 diabetes. Clinica Chimica Acta 399:112–116CrossRefGoogle Scholar
  4. Carlson JJ, Turpin AA, Wiebke G, Hunt SC (2009) Pre- and post-prandial appetite hormone levels in normal weight and severely obese women. Nutr Metab (Lond) 11(6):32–37CrossRefGoogle Scholar
  5. Castaneda TR, Tong J, Datta R, Culler M, Tscop MH (2010) Ghrelin in the regulation of body weight and metabolism. Front Neuroendocrinol 31:44–60PubMedCrossRefGoogle Scholar
  6. Chevenne D, Trivin F, Porquet D (1999) Insulin assays and reference values. Diabetes Metab 25(6):459–476PubMedGoogle Scholar
  7. Choi HJ, Cho YM, Moon MK, Choi HH, Shin HD, Jang HC, Kim SY, Lee HK, Park KS (2006) Polymorphisms in the ghrelin gene are associated with serum high-density lipoprotein cholesterol level and not with type 2 diabetes mellitus in Koreans. J Clin Endocrinol Metab 91:4657–4663PubMedCrossRefGoogle Scholar
  8. Dezaki K, Sone H, Yada T (2008) Ghrelin is a physiological regulator of insulin release in pancreatic islets and glucose homeostasis. Pharmacol Ther 118(2):239–249PubMedCrossRefGoogle Scholar
  9. Fossati P, Prencipel L (1982) Estimation of triglycerides concentration in serum and plasma. Clin Chem 28:2077–2080PubMedGoogle Scholar
  10. Friedwald WT, Levy RI, Fredrickson DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma without use of preparative ultracentrifuge. Clin Chem 18:499–502Google Scholar
  11. Granata R, Settanni F, Biancone L, Trovato L, Nano R, Bertuzzi F, Destefanis S, Annunziata M, Martinetti M, Catapano F, Ghè C, Isgaard J, Papotti M, Ghigo E, Muccioli G (2007) Acylated and unacylated ghrelin promote proliferation and inhibition of apoptosis of pancreatic β-cells and human islets. Endocrinology 148(2):512–529PubMedCrossRefGoogle Scholar
  12. Hung T, Sievenpiper JL, Marchie A, Kendall CW, Jenkins DJ (2003) Fat versus carbohydrates in insulin resistance, obesity, diabetes and cardiovascular disease. Curr Opin Clin Nutr Metab Care 6(2):165–176PubMedCrossRefGoogle Scholar
  13. Ikezaki A, Hosoda A, Ito K, Iwama S, Miura N, Matsouka H, Konden C, Kojima M, Kangawa K, Sugihara S (2002) Fasting plasma ghrelin levels are negatively correlated with insulin resistance and PAI-1, but not with leptin in obese children and adolescents. Diabetes 51:3408–3411PubMedCrossRefGoogle Scholar
  14. Karczewska-Kupczewska M, Straczkowski M, Adamska A, Nikolajuk A, Otziomek E, Górska M, Kowalska I (2010) Increased suppression of serum ghrelin concentration by hyperinsulinemia in women with anorexia nervosa. Eur J Endocrinol 162(2):235–239PubMedCrossRefGoogle Scholar
  15. Kim C, Robert B, David G, Mangione C, Selby J, Herman W (2005) Is physician gender associated with the quality of diabetes care? Health services, psychosocial research. Diabetes Care 28:1594–1598PubMedCrossRefGoogle Scholar
  16. Knapp RG, Miller MC (1992) Describing the performance of a diagnostic test. Clinical epidemiology and biostatistics. Williams & Wilkins, Baltimore, p 42Google Scholar
  17. Kunst A, Draeger B, Ziegenhorn J (1984) Methods of enzymatic analysis (Bergmeyer, H.U., Hrsg.) 3rd ed. Verlag, Weinheim. vol. VI, pp. 163–172Google Scholar
  18. Kuzuya M, Ando F, Iguchi A, Shimokata H (2006) Preproghrelin Leu72Met variant contributes to overweight in middle-aged men of a Japanese large cohort. Int J Obes 30:1609–1614CrossRefGoogle Scholar
  19. Larsen LH, Gjesing AP, Sørensen TI, Hamid YH, Echwald SM, Toubro S, Black E, Astrup A, Hansen T, Pedersen O (2005) Mutation analysis of the preproghrelin gene: no association with obesity and type 2 diabetes. Clin Biochem 38:420–424PubMedCrossRefGoogle Scholar
  20. Little RR, England JD, Wiedmeyer HH, Goldstein DE (1983) Effects of whole blood storage on results of glycosylated hemoglobin as measured by ion exchange chromatography, affinity chromatography and colorimetric. Clin Chem 29:1113–1115PubMedGoogle Scholar
  21. Mager U, Kolehmainen M, Lindström J, Eriksson JG, Valle TT, Hämäläinen H, Ilanne-Parikka P, Keinänen-Kiukaanniemi S, Tuomilehto JO, Pulkkinen L, Uusitupa MI (2006) Association between ghrelin gene variations and blood pressure in subjects with impaired glucose tolerance. Am J Hypertens 19:920–926PubMedCrossRefGoogle Scholar
  22. Matthews DR, Hosker JP, Rudenski AS (1985) Homeostasis model assessment: insulin resistance and β-cell function from fasting plasma glucose and insulin concentration in man. Diabetologia 28:412–419PubMedCrossRefGoogle Scholar
  23. Pacifico L, Poggiogalle E, Costantino F, Anania C, Ferraro F, Chiarelli F, Chiesa C (2009) Acetylated and non-acetylated ghrelin levels and their association with insulin resistance in obese and normal weight children with metabolic syndrome. Eur J Endocrinol 161(6):861–870PubMedCrossRefGoogle Scholar
  24. Porstmann T, Kiessig ST (1992) Enzyme immunoassay techniques. An overview. J Immunol Methods 150:5–21PubMedCrossRefGoogle Scholar
  25. Pulkkinen L, Ukkola O, Kolehmainen M, Uusitupa M (2010) Ghrelin in diabetes and metabolic syndrome. Int J Pept: pii: 248948Google Scholar
  26. Rantela GS, Liedtke RJ (1978) Automated enzymatic measurement of total cholesterol in serum. Clin Chem 24:112–115Google Scholar
  27. Rifai N, Warnick GR, Dominiczak MH (1997) Measurement of high density lipoprotein cholesterol. In Handbook of lipoprotein testing, Washington AACC Press pp. 221–245Google Scholar
  28. St-Pierre DH, Karelis AD, Coderre L, Malita F, Fontaine J, Mignault D (2007) Association of acylated and non-acylated ghrelin with insulin sensitivity in overweight and obese postmenopausal women. J Clin Endocrinol Metab 92:264–269PubMedCrossRefGoogle Scholar
  29. Takezawa J, Yamada K, Morita A, Aiba N, Watanabe S (2009) Preproghrelin gene polymorphisms in obese Japanese: association with diabetes mellitus in men and with metabolic syndrome parameters in women. Ob Res Clin Prac 3:179–191CrossRefGoogle Scholar
  30. Tang NP, Wang LS, Yang L, Gu HJ, Zhu HJ, Zhou B, Sun QM, Cong RH, Wang B (2008) Preproghrelin Leu72Met polymorphism in Chinese subjects with coronary artery disease and controls. Clin Chim Acta 387:42–47PubMedCrossRefGoogle Scholar
  31. Ukkola O (2009) Ghrelin and metabolic disorders. Curr Prot Pept Sci 10(1):2–7CrossRefGoogle Scholar
  32. Ukkola O, Ravussin E, Jacobson P, Pérusse L, Rankinen T, Tschöp M, Heiman ML, Leon AS, Rao DC, Skinner JS et al (2002) Role of ghrelin polymorphisms in obesity based on three different studies. Obes Res 10:782–791PubMedCrossRefGoogle Scholar
  33. Van der Lely A (2009) Ghrelin and new metabolic frontiers. Horm Res 71(1):129–133PubMedCrossRefGoogle Scholar
  34. Vartiainen J, Rajala U, Jokelainen J, Keinanen-Kiukaanniemi S, Kesaniemi YA, Ukkola O (2010) Serum ghrelin and the prediction of the development of impaired glucose regulation and type 2 diabetes in middle-aged subjects. J Endocrinol Invest 33:496–500PubMedGoogle Scholar
  35. Vivenza D, Rapa A, Castellino N, Bellone S, Petri A, Vacca G, Aimaretti G, Broglio F, Bona G (2004) Ghrelin gene polymorphisms and ghrelin, insulin, IGF-I, eptin and anthropometric data in children and adolescents. Eur J Endocrinol 151:127–133PubMedCrossRefGoogle Scholar
  36. Weickert MO, Loeffelholz CV, Arafat AM (2008) Euglycemic hyperinsulinemia differentially modulates circulating total and acylated ghrelin in humans. J Endocrinol Invest 31(2):119–124PubMedGoogle Scholar
  37. Williams DL, Cummings DE (2005) Regulation of ghrelin in physiologic and pathophysiologic states. J Nutr 135:1320–1325PubMedGoogle Scholar
  38. World Medical Association (2008) Declaration of Helsinki. Ethical principles for medical research involving human subjects, the 59th WMA General Assembly Seoul, South KoreaGoogle Scholar
  39. Wren AM, Seal LJ, Cohen MA, Brynes AE, Frost GS, Murphy KG, Dhillo WS, Ghatei MA, Bloom SR (2001) Ghrelin enhances appetitie and increases food intake in humans. J Clin Endocrinol Metab 86(12):5992PubMedCrossRefGoogle Scholar
  40. Xu LL, Xiang HD, Qiu CC, Xu Q (2008) Association of ghrelin polymorphisms with metabolic syndrome in Han Nationality Chinese. Biomed Environ Sci 21:188–192PubMedCrossRefGoogle Scholar
  41. Yada T, Dezaki K, Sone H (2008) Ghrelin regulates insulin release and glycemia: physiological role and therapeutic potential. Curr Diabetes Rev 4(1):18–23PubMedCrossRefGoogle Scholar
  42. Zavarella S, Petrone A, Zampetti S, Gueorguiev M, Spoletini M, Mein CA, Leto G, Korbonits M, Buzzetti R (2008) A new variation in the promoter region, the −604 CNT, and the Leu72Met polymorphism of the ghrelin gene are associated with protection to insulin resistance. Int J Obes (Lond) 32(4):663–668CrossRefGoogle Scholar
  43. Zou CC, Huang K, Liang L, Zhao ZY (2008) Polymorphisms of the ghrelin/obestatin gene and ghrelin levels in Chinese children with short stature. Clin Endocrinol (Oxf) 69:99–104CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Limited 2011

Authors and Affiliations

  • Nadida A. Gohar
    • 1
  • Dina F. Elgayar
    • 1
    • 4
  • Ayat I. Hassan
    • 2
  • Samar H. Aboulsoud
    • 3
  • Mona A. Hegazy
    • 3
  1. 1.Department of Clinical and Chemical Pathology, Faculty of MedicineCairo UniversityCairoEgypt
  2. 2.Department of Clinical Pathology, National Institute of Diabetes and EndocrinologyCairo UniversityCairoEgypt
  3. 3.Department of Internal Medicine, Faculty of MedicineCairo UniversityCairoEgypt
  4. 4.GizaEgypt

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