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V Sibilia
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AE Rigamonti
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F Pagani
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N Lattuada
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F Guidobono
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WB Wehrenberg
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EE Muller
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C Netti
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The effects of neonatal passive immunization against GHRH on bone was examined in male and female rats. Pups were treated subcutaneously with GHRH-antiserum (GHRH-Ab) from day 1 to day 10 of age. Bone mineral content (BMC) and bone mineral density (BMD) were evaluated at monthly intervals until 7 months. Markers of bone resorption (urinary lysylpyridinoline, LP), bone formation (serum osteocalcin, OC) and serum IGF-I were measured at 2, 3 and 7 months. In male rats, GHRH-Ab did not modify BMC and BMD when compared with controls. In contrast, female rats demonstrated lower whole body and femoral BMC and BMD from 2 to 7 months of age. Reduced bone growth in the females was associated with lower IGF-I levels than controls at 2 and 3 months of age, whereas in males IGF-I titers did not change during the period of the study. LP excretion was higher in GHRH-Ab-treated rats at 2 and 3 months in both sexes. In females, no difference in OC values was recorded, whereas in GHRH-Ab-treated males, there was an increase in OC levels at 2 and 3 months. These data indicate that transient GHRH deprivation induces an osteopenic effect in female rats which is not evident in male rats.

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E Mrak Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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I Villa Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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R Lanzi Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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M Losa Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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F Guidobono Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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A Rubinacci Bone Metabolic Unit,
Endocrine Unit, Scientific Institute San Raffaele, Via Olgettina, 60, 20132 Milano, Italy
Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

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It is presently thought that osteoprotegerin (OPG) is a cytokine involved in the regulation of osteoblast/osteoclast crosstalk and maintenance of bone mass. Recent studies showed that GH replacement therapy in GH-deficient patients was able to induce a significant increase of OPG in the plasma, as well as in the cortical and the trabecular bone. In order to determine whether GH could directly modulate OPG secretion, the effect of GH on human osteoblast-like cells (hOB) in primary culture was studied. After detecting the presence of the mRNA for the GH receptor (GHR) by RT-PCR, hOB were exposed to increasing concentrations of GH, from 0.1 to 25 ng/ml, for 24 h. The results showed that GH exposure was able to stimulate OPG secretion in a concentration-dependent manner. In addition, the OPG mRNA levels were increased, indicating that the hormone has a stimulatory effect on gene expression. The stimulatory effect on OPG expression and production was prevented by exposing the cells to tyrphostin AG490 (10 μM), an inhibitor of Janus kinase 2, which is one of the kinases involved in the intracellular pathway activated by the binding of GH to its receptor. Similar results were obtained when the cells were exposed to a receptor antagonist of GH, pegvisomant at 50 nM. GH exposure neither induced an increase in IGF-I expression nor secretion in hOB. These results suggest that the stimulation of OPG production induced by GH in hOB is specific and receptor mediated and further support the view that GH is able to modulate bone remodeling by directly influencing osteoblast–osteoclast crosstalk.

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