title: Osthol, a coumarin isolated from common cnidium fruit, enhances the differentiation and maturation of osteoblasts in vitro.
abstract: The effect of osthol on osteoblasts was investigated in primary osteoblastic cells isolated from newborn Wistar rats. Osthol was supplemented into cultured medium at 10⁻⁷, 10⁻⁶, 10⁻⁵ and 10⁻⁴ mol/l, respectively. No stimulating effect was found on cell proliferation, but 10⁻⁵ mol/l osthol caused a significant increase in alkaline phosphatase (ALP) activity. Osteogenic differentiation markers were examined over a period of time at this concentration, and compared with control cells that were not supplemented with osthol. The results showed that the ALP activity, osteocalcin secretion and calcium deposition level in cells treated with osthol were 1.52, 2.74 and 2.0 times higher, respectively, than in the control cells. Results of ALP histochemical staining and mineralized bone nodule assays both showed that the number and area achieved in osthol-treated cells were 1.53-fold higher than in control cells. The gene expression of the growth and transcription factors basic fibroblast growth factor, insulin-like growth factor I, bone morphogenetic protein 2 (BMP-2), runt-related gene 2 (Runx-2) and osterix, which are associated with bone development, were also investigated. The increase in mRNA expression was 1.94, 1.74, 1.68, 1.83 and 2.31 times, respectively, higher compared to the control. Furthermore, osthol increased the protein expression of p38 mitogen-activated protein kinase (MAPK) and type I collagen. p38MAPK protein and collagen in osthol-treated cells were 1.42 and 1.58 times higher in osthol-treated cells compared to the control. The results of these studies support the conclusion that osthol significantly enhances the osteogenic differentiation of cultured osteoblasts. The results also indicated that osthol could stimulate the osteoblastic differentiation of rat calvarial osteoblast cultures by the BMP-2/p38MAPK/Runx-2/osterix pathway and that osthol may be used as an important compound in the development of new antiosteoporosis drugs.
authors: Institute of Orthopaedics, Lanzhou General Hospital, Lanzhou Command of CPLA, Lanzhou, China.
authors: Lei-Guo
authors: LG
authors: Ming
authors: Jian
authors: J
authors: Zhou
authors: Guo-Zheng
authors: GZ
authors: Cheng
authors: Hui-Ping
authors: HP
authors: Ma
authors: Ke-Ming
authors: KM
authors: Chen
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis / Q000502:physiology; D000818:Animals; D050071:Bone Density Conservation Agents / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002113:Calcification, Physiologic / Q000502:physiology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation; D031048:Cnidium / Q000737:chemistry*; D003094:Collagen / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D003374:Coumarins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D005347:Fibroblasts / Q000502:physiology; D005638:Fruit; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D023421:Models, Animal; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D015675:Osteocalcin / Q000378:metabolism; D010024:Osteoporosis / Q000188:drug therapy / Q000517:prevention & control*; D008517:Phytotherapy; D028321:Plant Preparations / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012886:Skull / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D050071:Bone Density Conservation Agents; D055396:Bone Morphogenetic Protein 2; D002121:Calcium Channel Blockers; D024042:Collagen Type I; D003374:Coumarins; D028321:Plant Preparations; D016212:Transforming Growth Factor beta; D015675:Osteocalcin; D007334:Insulin-Like Growth Factor I; D003094:Collagen; D048051:p38 Mitogen-Activated Protein Kinases; D000469:Alkaline Phosphatase; C046627:osthol