Influence of ovariectomy on bone metabolism in very old rats.

Twenty-five 30-month-old Lou rats fed a diet (6 g/100 g BW/day) containing 0.9% Ca and 0.8% Pi were divided into five groups. Four groups were surgically ovariectomized. From day 2 until day 29 after ovariectomy, they were S.C. injected either with 17 beta estradiol (E2; 10 micrograms/kg BW/48 hours) or progesterone (P; 140 micrograms/kg BW/48 hours), or 17 beta estradiol + progesterone (E2P) at the same doses, or solvent alone (OVX). The fifth group was sham operated (SH) and injected with solvent. Urine was collected in metabolic cages from day 24 to 29 after ovx, and urinary pyridinoline (PYD) and deoxypyridinoline (DPD) excretion (markers of bone resorption) was measured by HPLC. All animals were killed 30 days after ovariectomy. Serum was then collected for measurement of osteocalcin (OC), alkaline phosphatase (ALP), parathyroid hormone (PTH), and calcitonin (CT). At necropsy, the success of ovariectomy was checked by marked atrophy of the uterine horns. Left and right femur were harvested for densitometric and mineral analysis, respectively. Ovariectomy had no significant effect upon plasma calcium and PTH concentrations. E2 or E2P treatment significantly increased plasma PTH and calcitonin concentrations. Plasma OC concentrations and ALP were not different in any of the groups. In contrast, urinary excretion of PYD and DPD was higher in OVX than in SH rats. Bone mineral density (BMD) of the distal femur was decreased by OVX, but was not different in the E2P and SH groups. A similar pattern was observed for the mineral or Ca content of whole femur. Thus, OVX decreased BMD and bone mineral content (BMC) in very old female rats. Plasma OC concentration and ALP activity failed to demonstrate any significant effect of OVX, whereas PYD and DPD were elevated. These results suggest that bone resorption is increased in OVX rats, even when supplemented with E2 or P alone. However, no significant difference was observed between SH and OVX rats treated with supplementation of both E2 and P. Thus, in very old rats, a combination of E2 and P is much more effective than E2 or P alone to prevent bone loss following ovariectomy.

[Parathyroid hormone related peptide (PTHrP) and bone metabolism]. / Peptide apparenté à l'hormone parathyroïdienne (PTHrP) et métabolisme osseux.

The discovery of PTHrP was the result of research on the mechanisms by which some cancers cause hypercalcemia (humoral hypercalcemia of malignancy) without necessarily metastasizing to bone. PTHrP is also present in various normal adult and fetal tissues. Its concentration is normally very low (picomolar) in blood, but it is more abundant in milk (nanomolar concentration). PTHrP seems able to exert autocrine/paracrine as well as endocrine effects on bone metabolism. A major role for PTHrP in regulation of fetal bone metabolism has been demonstrated in mice. Homologous recombination has been used in these rodents to remove the major coding exon from one copy of the mouse PTHrP gene in embryonic stem cells. Subsequently generated chimeric mice transmit the mutant PTHrP allele through the germline. Homozygous mutants died immediately after birth and had a multitude of skeletal abnormalities. So PTHrP seems necessary to embryonic development of the skeleton. PTHrP (1-34), like PTH (1-34) fragments, might be responsible for both bone resorption and formation. Although the effects of the carboxyl-terminal fragments are still controversial, PTHrP (107-111) fragment seems able to inhibit osteoclast activity. PTHrP (1-34), whose 8 of the first 13 amino-acids are identical with those in PTH (1-34), acts through the same receptor as PTH on osteoblasts and renal cells membrane. The PTHrP/PTH receptor sequence is now well established. PTHrP-receptor coupling is mediated by cyclic AMP and/or inositols-phosphate. The consequent activation of protein kinase A and intracellular calcium or protein kinase C, respectively, locally induces growth factors or cytokines secretion, responsible for the observed effects. The role of PTHrP appears important during pregnancy and lactation, when it stimulates fetal bone growth by increasing calcium transport from the dam to its fetus and maternal bone resorption allowing calcium supply for milk production, respectively. Such a role would be particularly important in domestic ruminants, which are often simultaneously pregnant and lactating. The role of PTHrP during aging (especially in post-menopausal women in which bone loss may induce osteoporosis) remains unknown and might be of peculiar interest since PTHrP (1-34) and (107-111) are able to restore bone loss induced by ovariectomy in rats.

Short communication: parathyroid hormone-related peptide does not stimulate phosphate placental transport.

It has been demonstrated that parathyroid hormone-related peptide (PTHrP) stimulates calcium (Ca) placental transport. Ca and inorganic phosphorous (P) metabolisms are intimately linked. Thus we have studied the influence of PTHrP on P placental transport in ewes. In our experimental paradigm, synthetic human PTHrP (1-86) fragment intravenously injected in six single chronically catheterized fetuses (3 x 1 micrograms/fetus/day, from day 129 to day 140 of gestation) had no significant effect upon P placental transport which was 131 +/- 6 and 129 +/- 9 mg/kg fetal weight/day in treated and control fetuses, respectively.

Preserved bone mass in ovariectomized rats treated with parathyroid-hormone-related peptide (1-34) and (107-111) fragments.

The effect of synthetic human PTHrP (1-34) or (107-111) fragments on bone loss was studied in rats, one month after ovariectomy (OVX). Four groups of 7-8-month-old rats were treated sc daily for 13 d with PTHrP (1-34) or (107-111) at the dose of 1 or 3 nmol/100 g body weight. Sham-operated (SHO) and control OVX rats received solvent alone. In our conditions, at the lowest dose, neither (1-34) nor (107-111) fragments had any significant effect. However, at the dose of 3 nmol/100 g daily for 13 d both treatments significantly increased femoral dry weight, ash weight, Ca content and densitometry of the femur. The effect of PTHrP (1-34) mainly resulted from increased cortical and trabecular bone (% recovery: 98.25 and 105.23%, respectively). For the PTHrP (107-111) fragment, a positive effect was only demonstrated on the cortical bone (98.25% recovery). The results of this study demonstrate that both hPTHrP (1-34) and (107-111, osteostatin) fragments are positive for bone when administered at the dose of 3 nmol/100 g body weight/d for 13 d to adult OVX rats.

Parathyroid hormone-related peptide may increase mammary blood flow.

Amino-terminal fragments of PTHrP were previously shown to increase regional blood flow in laboratory animals. Since PTHrP is produced in the lactating mammary gland and associated nutrient vessels, we examined the effects of peptide fragments of PTHrP on the hemodynamics of the mammary gland of dried sheep. The left arterial mammary blood flow measured using ultrasonic flow probes in four dried Lacaune ewes was 233 +/- 11 ml/minute. It was significantly increased when synthetic human PTHrP-(1-34) or (1-86) fragments were injected into the mammary artery. The effect was dose dependent for PTHrP-(1-34), varying between 0.0075 and 0.3 nmol/kg body weight. PTHrP-(140-173) fragment lacked any vasorelaxant activity. Synthetic human endothelin (ET1) decreased arterial blood flow in a dose-dependent manner. This decrease was inhibited by PTHrP-(1-34), and this inhibition was PTHrP dose related. When ET1 (10 pmol/kg body weight) was injected together with PTHrP-(1-86) (100 pmol/kg body weight), only a significant increase in mammary blood flow was observed. Thus, PTHrP produced by the lactating mammary gland may be involved in the regulation of mammary blood flow.

Parathyroid hormone-related peptide and milking-induced phosphaturia in dairy cows.

Five intact and four thyroparathyroidectomized (TPTX) lactating dairy cows were fitted with polyvinyl catheters implanted into the left carotid artery and the left mammary vein (for blood sampling) and into the right external jugular vein (for injections), and with a Foley catheter inserted into the urinary bladder. In these nine cows, daily morning and evening milking induced a prompt and transient rise in mammary venous blood plasma parathyroid hormone-related peptide (PTHrP)(1-34) concentrations, followed by an increase in both urinary phosphate concentration and phosphate renal clearance. Such a milking-induced phosphaturia was not observed when (tyr)34-bPTH(7-34)-NH2 (5 pmol/kg body wt) was infused during milking into the TPTX cows. These results indicate that PTHrP released systematically by the mammary gland during milking might be responsible for the observed phosphaturia.