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Journal of Clinical Endocrinology & Metabolism , doi:10.1210/jc.2007-2235
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The Journal of Clinical Endocrinology & Metabolism Vol. 93, No. 5 1980-1987
Copyright © 2008 by The Endocrine Society

Novel Ghrelin Assays Provide Evidence for Independent Regulation of Ghrelin Acylation and Secretion in Healthy Young Men

Jianhua Liu1, Catherine E. Prudom1, Ralf Nass, Suzan S. Pezzoli, Mary C. Oliveri, Michael L. Johnson, Paula Veldhuis, David A. Gordon, Andrew D. Howard, Derrick R. Witcher, H. Mario Geysen, Bruce D. Gaylinn and Michael O. Thorner

Departments of Medicine (J.L., R.N., S.S.P., M.C.O., B.D.G., M.O.T.), Chemistry (C.E.P., H.M.G.), and Pharmacology (M.L.J., P.V.), University of Virginia, Charlottesville, Virginia 22908; Bristol-Myers Squibb Pharmaceutical Research Institute (D.A.G.), Princeton, New Jersey 08543; Merck Research Laboratories (A.D.H.), Rahway, New Jersey 07065; and Lilly Research Laboratories (D.R.W.), Indianapolis, Indiana 46285

Address all correspondence and requests for reprints to: Michael O. Thorner, University of Virginia Health System, Endocrinology and Metabolism, Box 801411, Charlottesville, Virginia 22908. E-mail: MOT{at}virginia.edu.

Context: Ghrelin, an acylated peptide hormone secreted from the gut, regulates appetite and metabolism. Elucidating its pattern of secretion in the fed and fasted states is important in the face of the obesity epidemic.

Objective: Our objective was to examine changes in circulating ghrelin and des-acyl ghrelin in response to meals and fasting using newly developed two-site sandwich assays and sample preservation protocols to allow specific detection of full-length forms.

Design: Ten-minute sampling was done for 26.5 h during a fed admission with standardized meals and on a separate admission during the final 24 h of a 61.5-h fast and continuing for 2.5 h after terminating the fast.

Setting: The study was conducted at the University Hospital General Clinical Research Center.

Participants: Eight male volunteers participated, mean ± SD age 24.5 ± 3.7 yr and body mass index 24 ± 2.1 kg/m2.

Main Outcome Measures: Ten-minute sampling profiles were assessed for ghrelin and des-acyl ghrelin, fed and fasting.

Results: In the fed state, ghrelin and des-acyl ghrelin showed similar dynamics; both were sharply inhibited by meals and increased at night. During fasting, ghrelin decreased to nadir levels seen postprandially, and des-acyl ghrelin remained near peak levels seen preprandially. Total full-length ghrelin (acyl plus des-acyl) levels remained unchanged.

Conclusions: Meals inhibited secretion of both ghrelin and des-acyl ghrelin, yet long-term fasting inhibited acylation but not total secretion. Acylation may be regulated independently of secretion by nutrient availability in the gut or by esterases that cleave the acyl group. These studies highlight the importance of stringent conditions for sample collection and evaluation of full-length ghrelin and des-acyl ghrelin using specific two-site assays.




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