title: Puerarin enhances proliferation and osteoblastic differentiation of human bone marrow stromal cells via a nitric oxide/cyclic guanosine monophosphate signaling pathway.
abstract: Puerarin, a major active isoflavone extracted from the Traditional Chinese Medicine Radix Puerariae, has been studied for its comprehensive biological effects. However, to date, its effect on bone formation and the underlying mechanism of action have not been well investigated. The present study investigated the effect of puerarin on cell proliferation and osteoblastic maturation in cultured human bone marrow stromal cells (hBMSC) in vitro. Puerarin (2.5-100 µM) increased hBMSC growth in a dose-dependent manner, as indicated by an MTT assay, and stimulated osteoblastic maturation as indicated by assessment of alkaline phosphatase (ALP) activity, as well as calcium deposition into the extracellular matrix detected by alizarin red S staining. Furthermore, polymerase chain reaction analysis showed that the expression of osteoblastic markers, including Runt-related transcription factor 2/core-binding factor alpha 1, osterix and osteocalcin, were increased in hBMSCs following incubation with puerarin. Further experiments indicated that puerarin increased the nitric oxide (NO) production and cyclic guanosine monophosphate (cGMP) content in hBMSCs. The effects of puerarin were mimicked by 17β-estrodiol (10(-8) M) and were abolished in the presence of estrogen receptor antagonist ICI182780 (10(-7) M). A NO synthase inhibitor, Nx-nitro-L-arginine methylester (6 x 10(-3) M), significantly attenuated puerarin-induced increases in NO production and cGMP content, in parallel with a reduction of cell proliferation and osteoblastic differentiation as well as the expression of osteoblastic markers. These results suggested that puerarin may prevent osteoporosis by exerting stimulatory effects on bone formation and the NO/cGMP pathway, which has an important role in puerarin-induced hBMSC proliferation and osteoblastic differentiation.
authors.affiliation: Department of Endocrinology and Metabolism, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Haihong
authors.initials: H
authors.lastname: Lv
authors.affiliation: Baiyuan Gene Technology Co. Ltd, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Tuanjie
authors.initials: T
authors.lastname: Che
authors.affiliation: Department of Endocrinology and Metabolism, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Xulei
authors.initials: X
authors.lastname: Tang
authors.affiliation: Department of Endocrinology and Metabolism, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Endocrinology and Metabolism, The First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Cheng
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D005260:Female; D000077267:Fulvestrant; D005786:Gene Expression Regulation; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D008297:Male; D008875:Middle Aged; D009569:Nitric Oxide / Q000819:agonists / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D019335:Nitroarginine / Q000494:pharmacology; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D015398:Signal Transduction; D000076182:Sp7 Transcription Factor; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D007529:Isoflavones; C493709:RUNX2 protein, human; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors; D015675:Osteocalcin; D019335:Nitroarginine; D000077267:Fulvestrant; D009569:Nitric Oxide; D004958:Estradiol; D000469:Alkaline Phosphatase; D006152:Cyclic GMP; D002118:Calcium; C033607:puerarin