title: Glucocorticoid-induced bone loss in mice can be reversed by the actions of parathyroid hormone and risedronate on different pathways for bone formation and mineralization.
abstract: OBJECTIVE
  Glucocorticoid excess decreases bone mineralization and microarchitecture and leads to reduced bone strength. Both anabolic (parathyroid hormone [PTH]) and antiresorptive agents are used to prevent and treat glucocorticoid-induced bone loss, yet these bone-active agents alter bone turnover by very different mechanisms. This study was undertaken to determine how PTH and risedronate alter bone quality following glucocorticoid excess.
  
  
  METHODS
  Five-month-old male Swiss-Webster mice were treated with the glucocorticoid prednisolone (5 mg/kg in a 60-day slow-release pellet) or placebo. From day 28 to day 56, 2 groups of glucocorticoid-treated animals received either PTH (5 microg/kg) or risedronate (5 microg/kg) 5 times per week. Bone quality and quantity were measured using x-ray tomography for the degree of bone mineralization, microfocal computed tomography for bone microarchitecture, compression testing for trabecular bone strength, and biochemistry and histomorphometry for bone turnover. In addition, real-time polymerase chain reaction (PCR) and immunohistochemistry were performed to monitor the expression of several key genes regulating Wnt signaling (bone formation) and mineralization.
  
  
  RESULTS
  Compared with placebo, glucocorticoid treatment decreased trabecular bone volume (bone volume/total volume [BV/TV]) and serum osteocalcin, but increased serum CTX and osteoclast surface, with a peak at day 28. Glucocorticoids plus PTH increased BV/TV, and glucocorticoids plus risedronate restored BV/TV to placebo levels after 28 days. The average degree of bone mineralization was decreased after glucocorticoid treatment (-27%), but was restored to placebo levels after treatment with glucocorticoids plus risedronate or glucocorticoids plus PTH. On day 56, RT-PCR revealed that expression of genes that inhibit bone mineralization (Dmp1 and Phex) was increased by continuous exposure to glucocorticoids and glucocorticoids plus PTH and decreased by glucocorticoids plus risedronate, compared with placebo. Wnt signaling antagonists Dkk-1, Sost, and Wif1 were up-regulated by glucocorticoid treatment but down-regulated after glucocorticoid plus PTH treatment. Immunohistochemistry of bone sections showed that glucocorticoids increased N-terminal Dmp-1 staining while PTH treatment increased both N- and C-terminal Dmp-1 staining around osteocytes.
  
  
  CONCLUSIONS
  Our findings indicate that both PTH and risedronate improve bone mass, degree of bone mineralization, and bone strength in glucocorticoid-treated mice, and that PTH increases bone formation while risedronate reverses the deterioration of bone mineralization.
authors.affiliation: University of California Davis Medical Center, Sacramento.
authors.forename: Wei
authors.initials: W
authors.lastname: Yao
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Cheng
authors.forename: Aaron
authors.initials: A
authors.lastname: Pham
authors.forename: Cheryl
authors.initials: C
authors.lastname: Busse
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Zimmermann
authors.forename: Robert O
authors.initials: RO
authors.lastname: Ritchie
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Lane
mesh_terms: D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects*; D012968:Etidronic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D015870:Gene Expression; D005938:Glucocorticoids / Q000494:pharmacology*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D015675:Osteocalcin / Q000097:blood; D010012:Osteogenesis / Q000187:drug effects*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D011239:Prednisolone / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000068296:Risedronic Acid; D014057:Tomography, X-Ray Computed; D051153:Wnt Proteins / Q000502:physiology
chemical_list: D005938:Glucocorticoids; D010281:Parathyroid Hormone; D051153:Wnt Proteins; D015675:Osteocalcin; D011239:Prednisolone; D000068296:Risedronic Acid; D012968:Etidronic Acid