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Third Division, Department of Medicine, Kobe University School of Medicine Kobe 650, Japan
Address correspondence and requests for reprints to: Simeon I. Taylor, M.D., Ph.D., National Institutes of Health, Building 10, Room 8S-243, Bethesda, Maryland 20892.
A sensitive RIA for human GH-releasing hormone-(l–44)-NH2 [hGHRH-(l–44)-NH2] was developed which allows its measurement in human plasma extracts. The assay did not detect hGHRH-(l–37)-OH or hGHRH-(l–40)-OH. A method to extract hGHRH from plasma was developed using silicic acid and acid-acetone, by which recovery of synthetic hGHRH-(l–44)-NH2 from plasma averaged 74.3%. Serial dilutions of plasma extracts gave an inhibition curve parallel with that of synthetic hGHRH-(l–44)-NH2 in the RIA system. On Sephadex G-50 columns, hGHRH-like immunoreactivity (hGHRH-LI) in plasma extracts eluted as a single peak corresponding to hGHRH-(l–44)-NH2. This hGHRH-LI peak, when subjected to reverse phase HPLC, emerged at the position where hGHRH-(l–44)-NH2 was eluted. hGHRH-LI was detectable in the peripherally circulating plasma of all subjects tested. The mean basal level of plasma hGHRH-LI in normal subjects was 9.4 ± 0.7 (±SE) pg/ml (n = 22; range, 2.8–18.1 pg/ml), comparable to the basal plasma hGHRH-LI concentration in patients with hypothalamic lesions (11.3 ± 1.1 pg/ml; n = 7). Oral administration of L-dopa (0.5 g) caused a significant increase in both plasma hGHRH-LI and GH levels in normal subjects, and the plasma hGHRH-LI peak slightly preceded or coincided with that of plasma GH in individual subjects. There was also a significant correlation between plasma hGHRH-LI and the GH rises after L-dopa administration when their net increments were compared. All of the patients with hypothalamic lesions had significant increases in plasma GH after hGHRH-(l–44)- NH2 injection (1 µg/kg BW, iv), indicating the presence of functioning somatotrophs in their pituitaries. When L-dopa was orally administered to these patients, neither plasma hGHRHLI nor GH concentration changed throughout a 120-min observation period.
These findings suggest that 1) hGHRH, immunologically and chromatographically indistinguishable from synthetic hGHRH-(l–44)-NH2, is detectable in peripheral plasma in humans; 2) Ldopa stimulates the release of hypothalamic hGHRH, alterations of which are reflected in changes in peripheral levels; and 3) the source of circulating hGHRH is not restricted to the hypothalamus, since hGHRH-LI is present in the peripheral plasma of patients with hypothalamic lesions in amounts similar to those found in normal subjects.
* This work was supported in part by research grantsfrom the Japanese Ministry of Health and Welfare; the Japanese Ministry of Education, Science, and Culture; and the Growth Science Foundation for 1984.
Received July 17, 1985.
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