Aim: Excessive apoptosis of osteoblasts is the major cause of low bone mass, and bovine lactoferrin (bLF), an iron-binding glycoprotein, might protect osteoblastic cells from apoptosis induced by serum withdrawal. The aim of this study was to elucidate the mechanisms underlying the anti-apoptotic action of bLF in rat osteoblasts in vitro.
Methods: Primary rat osteoblasts were incubated in the presence of varying concentrations of bLF for 24 h. The expression of insulin-like growth factor I (IGF-I) and IGF-I receptor (IGF-IR) was measured uisng RT-PCR and Western blotting. Cell apoptosis was examined with flow cytometry. siRNAs targeting IGF-I was used in this study.
Results: Treatment of bLF (0.1-1000 μg/mL) dose-dependently increased the expression of IGF-I and IGF-IR in the osteoblasts. Treatment with bLF (10, 100 μg/mL) markedly inhibited the osteoblast apoptosis (with the rate of total apoptosis of 70% at 10 μg/mL), but the high concentration of bLF (1000 μg/mL) significantly promoted the osteoblast apoptosis. Knockdown of the IGF-I gene in osteoblasts with siRNA markedly increased the osteoblast apoptosis.
Conclusion: Lactoferrin (10 and 100 μg/mL) effectively inhibits apoptosis of primary rat osteoblasts by upregulating IGF-I expression.