Osthole Enhances Osteogenesis in Osteoblasts by Elevating Transcription Factor Osterix via cAMP/CREB Signaling In Vitro and In Vivo

Abstract

Anabolic anti-osteoporotic agents are desirable for treatment and prevention of osteoporosis and fragility fractures. Osthole is a coumarin derivative extracted from the medicinal herbs Cnidium monnieri (L.) Cusson and Angelica pubescens Maxim.f. Osthole has been reported with osteogenic and anti-osteoporotic properties, whereas the underlying mechanism of its benefit still remains unclear. The objective of the present study was to investigate the osteopromotive action of osthole on mouse osteoblastic MC3T3-E1 cells and on mouse femoral fracture repair, and to explore the interaction between osthole-induced osteopromotive effect and cyclic adenosine monophosphate (cAMP) elevating effect. Osthole treatment promoted osteogenesis in osteoblasts by enhancing alkaline phosphatase (ALP) activity and mineralization. Oral gavage of osthole enhanced fracture repair and increased bone strength. Mechanistic study showed osthole triggered the cAMP/CREB pathway through the elevation of the intracellular cAMP level and activation of the phosphorylation of the cAMP response element-binding protein (CREB). Blockage of cAMP/CREB downstream signals with protein kinase A (PKA) inhibitor KT5720 partially suppressed osthole-mediated osteogenesis by inhibiting the elevation of transcription factor, osterix. In conclusion, osthole shows osteopromotive effect on osteoblasts in vitro and in vivo. Osthole-mediated osteogenesis is related to activation of the cAMP/CREB signaling pathway and downstream osterix expression.

Psoralen accelerates bone fracture healing by activating both osteoclasts and osteoblasts

“…For instance, root bark of Sambucus williamsii Hance, a plant from the family of Caprifoliaceae, promotes rat femoral fracture healing by promoting bone formation (41). Osthole, a bioactive coumarin derivative, promotes bone fracture healing by enhancing osteogenesis by osteoblasts (42, 43). Heart‐wood extract from Dalbergia sissoo promotes fracture healing by stimulating osteoblast function and inhibiting osteoclastic gene expression (44).…”

Osthole‐loaded N‐octyl‐O‐sulfonyl chitosan micelles (NSC‐OST) inhibits RANKL‐induced osteoclastogenesis and prevents ovariectomy‐induced bone loss in rats

“…In recent years, the anti‐osteoporotic effect of OST has gradually been expounded. It has been found that OST stimulates osteoblast differentiation and bone formation via the β‐catenin‐bone morphogenetic protein (BMP) signalling pathway or the cyclic adenosine 3′,5′‐monophosphate (cAMP)/response element‐binding protein (CREB) signalling network . Consistent with this stimulation, Zhao et al reported that OST inhibited OC production and prevented bone loss in an ovariectomized animal model of osteoporosis.…”

Phytochemistry, Ethnopharmacology, Pharmacokinetics and Toxicology of Cnidium monnieri (L.) Cusson

“…The bone morphogenetic protein was enhanced by cyclic adenosine monophosphate/cAMP response element-binding protein signaling pathway which targeted the transcription factor osterix. The expression of osterix was up-regulated by osthole at 20 mg/kg [173]. From in vitro studies, osthole (1 × 10 −6 mol/L and 1 × 10 −8 mol/L) upregulated osteoprotegerin (OPG) mRNA expression level but did not influence on receptor activator of NF-κB ligand (RANKL) mRNA expression level.…”

“…5 g/kg, 6 times per week In vivo [169] Anti-osteoporosis effect Total coumarins 2.5 mL/kg, 8 weeks via oral gavage In vivo [170] Anti-osteoporosis effect Imperatorin and Osthole Imperatorin: 0.1 µmol/L, Osthole: 10 µmol/L In vitro [171] Anti-osteoporosis effect Osthole 6.7 mg/kg, 6 times per week for 12 weeks In vivo [172] Anti-osteoporosis effect Osthole 200 mg/kg for 5 weeks via oral gavage In vivo [173] Anti-osteoporosis effect Osthole 20 mg/kg via local injection In vivo [174] Anti-osteoporosis effect Osthole 20 mg/kg for 7 days via oral gavage In vivo+ In vitro [175] Anti-osteoporosis effect Osthole 1 × 10 -6 and 1 × 10 −8 mol/L for 48 h and 72 h In vitro [175] Anti-osteoporosis effect Osthole 1 × 10 −1 , 1 × 10 −2 , 1 × 10 −3 , 1 × 10 −4 mol/l. In vitro [176] Anti-osteoporosis effect Osthole 1 × 10 −4 , 1 × 10 −5 , 1 × 10 −6 , 1 × 10 −7 mol/L.…”