The effect of His-D-Trp-Ala-Trp-D-Phe-Lys-NH
2(GHRP) on growth hormone (GH) release from cultured bovine anterior pituitary (AP) cells was studied in vitro with the interactive effects of GH-releasing factor (GRF: hpGRF (1-29)-NH
2) and somatostatin (SRIF). The AP cells (5×10
4cells per well) were incubated with media, and the media were changed 3 days after plating. After 3.5 days in culture, cells were incubated for 2h with the peptides. GHRP stimulated GH release from cultured cells in a dose-related manner. At doses from 10
-11to 10
-7M GHRP, the amount of GH released was significantly greater than the controls (
P<0.05 to
P<0.001). The amounts of GH released at lower doses of GHRP (10
-14to 10
-12M) were not significantly different from the controls. GH concentrations after treatment with 10
-11and 10
-7M GHRP were 3.98±0.27 and 4.81±0.16ng/ml, respectively. In experiments performed similarly, the 10
-7M GHRH, GHRP, and combined treatment with GHRP plus GHRH increased GH 126, 57, and 139%, respectively (
P<0.001). The GH releasing effects of either GHRH alone or GHRP plus GHRH were significantly more potent than that of GHRP alone (
P<0.001). The additive effect was not significant when compared with GHRH alone. GH release induced by either GHRH or GHRP was significantly inhibited by SRIF (
P<0.01) compared with the untreated control. The inhibitory effect of SRIF in combined treatment with GHRP plus GHRH was significantly less than that of SRIF with GHRH or with GHRP (
P<0.01). The present study suggests that GH-releasing peptide (GHRP) induced GH release in cattle via a direct action on anterior pituitary cells in vitro.
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