pTH (1-84) (rat)

Background: Intermittent PTH administration directly stimulates osteoblasts; however, mechanisms of bone accrual that are independent of the direct actions on osteoblasts may be under-appreciated. Our aims were to decipher (1) whether PTH 1-84 augments vasodilation of the femoral principal nutrient artery (PNA), (2) whether 15 days of intermittent PTH 1-84 augments endothelium-dependent and/or -independent vasodilation of the femoral PNA, and (3) the signaling mechanisms involved. Methods: Experiment 1: Femoral PNAs from male Wistar rats were exposed to cumulative doses of PTH 1-84 with and without an anti-vascular endothelial growth factor antibody and/or the endothelial NO synthase inhibitor l-NAME. Experiment 2: Male Wistar rats were administered PTH and/or the anti-VEGF antibody for 2 weeks. Subsequently, endothelium-dependent vasodilation to acetylcholine and endothelium-independent vasodilation to sodium nitroprusside were assessed. In addition, endothelium-dependent signaling pathways were analyzed by use of l-NAME and/or and the cyclooxygenase inhibitor indomethacin. Results: Cumulative doses of PTH 1-84 induced vasodilation of the femoral PNA, which was reduced by 38% and 87% with the anti-VEGF antibody and l-NAME, respectively. Secondly, 2 weeks of intermittent PTH 1-84 administration doubled trabecular bone volume, augmented bone formation parameters and reduced osteoclast activity. In addition, PTH enhanced endothelium-dependent vasodilation via up-regulation of NO. Co-administration of the anti-VEGF antibody (1) inhibited the PTH-induced increase in bone volume and remodeling parameters and (2) blunted the augmented vasodilator responsiveness of the PNA. Finally, endothelium-dependent vasodilation in PTH-treated rats was highly correlated with trabecular bone volume. Conclusion: As hypothesized, PTH enhanced endothelium-dependent vasodilation of the femoral PNA via augmented NO production and was mediated partially through VEGF signaling. Further, vasodilation to PTH appears independent of vascular smooth muscle cell participation. More importantly, the strong association between vasodilation and bone volume suggests that bone arteriolar function is critical for PTH-induced bone anabolism.

Although daily injections of parathyroid hormone (PTH) can rapidly reverse estrogen-deficiency bone loss in rats, PTH treatment of osteoporotic humans has to date produced more modest increases in bone mass. To explore the reasons for this important difference, we evaluated the dose- and time-dependence of human PTH 1-84 treatment effects on bone mass and biochemical markers of bone metabolism in rats with estrogen-deficiency bone loss. The highest doses of PTH increased spinal, femoral, and total skeletal mass to supra-normal levels and stimulated cortical endosteal bone formation. Spine and whole skeleton mass and density increased rapidly at first, but then increased more slowly; the rate of change decreased significantly (p < 0.01) during continued treatment with the highest doses of PTH. The effects of PTH treatment on biochemical markers also were both dose-dependent and time-dependent. Serum osteocalcin, a marker of osteoblast function, increased with the highest doses of PTH (p < 0.001), but reached an early plateau and later returned toward baseline. Urinary excretion of pyridinolines, a marker of osteoclast function, increased in a time-dependent fashion throughout treatment (p < 0.001). Serum 1,25(OH)2 vitamin D levels increased in a dose-related fashion, but then decreased toward control levels despite continued treatment. We demonstrate that both osteoblast and osteoclast function are increased during daily PTH therapy in the rat. The pattern of response depends on both the dose of PTH and the duration of therapy. These dose- and time-related effects should be taken into account when designing experimental PTH treatments for osteoporosis, and they deserve intensive study.

Parathyroid hormone (PTH) augments bone metabolism and bone mass when given intermittently. Enhanced blood flow is requisite to support high tissue metabolism. The bone arteries are responsive to all three PTH analogs, which may serve to augment skeletal blood flow during intermittent PTH administration. Introduction: PTH augments bone metabolism. Yet, mechanisms by which PTH regulates bone blood vessels are unknown. We deciphered (1) endothelium-dependent and endothelium-independent vasodilation to PTH 1-84, PTH 1-34, and PTHrP 1-34, (2) the signaling pathways (i.e., endothelial nitric oxide synthase [eNOS], cyclooxygenase [COX], protein kinase C [PKC], and protein kinase A [PKA]), and (3) receptor activation. Methods: Femoral principal nutrient arteries (PNAs) were given cumulative doses (10(-13)-10(-8) M) of PTH 1-84, PTH 1-34, and PTHrP 1-34 with and without signaling pathway blockade. Vasodilation was also determined following endothelial cell removal (i.e., denudation), PTH 1 receptor (PTH1R) inhibition and to sodium nitroprusside (SNP; a nitric oxide [NO] donor). Results: Vasodilation was lowest to PTH 1-34, and maximal dilation was highest to PTHrP 1-34. Inhibition of eNOS reduced vasodilation to PTH 1-84 (-80 %), PTH 1-34 (-66 %), and PTHrP 1-34 (-48 %), evidencing the contribution of NO. Vasodilation following denudation was eliminated (PTH 1-84 and PTHrP 1-34) and impaired (PTH 1-34, 17 % of maximum), highlighting the importance of endothelial cells for PTH signaling. Denuded and intact PNAs responded similarly to SNP. Both PKA and PKC inhibition diminished vasodilation in all three analogs to varying degrees. PTH1R blockade reduced vasodilation to 1, 12, and 12 % to PTH 1-84, PTH 1-34, and PTHrP 1-34, respectively. Conclusions: Vasodilation of femoral PNAs to the PTH analogs occurred via activation of the endothelial cell PTH1R for NO-mediated events. PTH 1-84 and PTHrP 1-34 primarily stimulated PKA signaling, and PTH 1-34 equally stimulated PKA and PKC signaling.