title: Lactoferrin promote primary rat osteoblast proliferation and differentiation via up-regulation of insulin-like growth factor-1 expression.
abstract: The aim of this study was to explore the effect of lactoferrin (LF) in primary fetal rat osteoblasts proliferation and differentiation and investigate the underlying molecular mechanisms. Primary rat osteoblasts were obtained from the calvarias of neonatal rats. Osteoblasts were treated with LF (0.1-1000 μg/mL), or OSI-906 [a selective inhibitor of insulin-like growth factor 1 (IGF-1) receptor and insulin receptor]. The IGF-1 was then knocked down by small hairpin RNA (shRNA) technology and then was treated with recombinant human IGF-1 or LF. Cell proliferation and differentiation were measured by MTT assay and alkaline phosphatase (ALP) assay, respectively. The expression of IGF-1 and IGF binding protein 2 (IGFBP2) mRNA were analyzed using real-time PCR. LF promotes the proliferation and differentiation of osteoblasts in a certain range (1-100 μg/mL) in time- and dose-dependent manner. The mRNA level of IGF-1 was significantly increased, while the expression of IGFBP2 was suppressed by LF treatment. Knockdown of IGF-1 by shRNA in primary rat osteoblast dramatically decreased the abilities of proliferation and differentiation of osteoblasts and blocked the proliferation and differentiation effect of LF in osteoblasts. OSI906 (5 μM) blocked the mitogenic and differentiation of LF in osteoblasts. Proliferation and differentiation of primary rat osteoblasts in response to LF are mediated in part by stimulating of IGF-1 gene expression and alterations in the gene expression of IGFBP2.
authors.affiliation: Endocrinology Department, Fujian Provincial Hospital, No. 134 Dong Jie Road, Fuzhou, 350001, China, hjm996@126.com.
authors.forename: Jian-ming
authors.initials: JM
authors.lastname: Hou
authors.forename: Man
authors.initials: M
authors.lastname: Wu
authors.forename: Qing-ming
authors.initials: QM
authors.lastname: Lin
authors.forename: Fan
authors.initials: F
authors.lastname: Lin
authors.forename: Ying
authors.initials: Y
authors.lastname: Xue
authors.forename: Xu-hua
authors.initials: XH
authors.lastname: Lan
authors.forename: En-yu
authors.initials: EY
authors.lastname: Chen
authors.forename: Mei-li
authors.initials: ML
authors.lastname: Wang
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Yang
authors.forename: Feng-xiong
authors.initials: FX
authors.lastname: Wang