Effects of sika pilose antler type I collagen on osteoclast and its molecular mechanism / 中草药

ABSTRACT

Objective: To explore the effect of sika pilose antler type I collagen (SPC-I) on osteoclast and its molecular mechanism. Methods: The osteoclasts and osteoblasts were cultured by the induction method of whole bone marrow cells. The control (with full medium), osteoclasts (with HG-DMEM inducing medium), and SPC-I (2.5, 5, and 10 g/L) groups were set up. Except the control group, others were given the HG-DMEM inducing medium with each 40 ng/mL of both RANKL and macrophage colony-stimulating factor (M-CSF), then conditioned cultured for 7 d, every other 3 d to replace medium for the complement of the drug concentration. By HE and tartrate-resistant acid phosphatase (TRAP) stainings, the cell morphology was observed under inverted microscope. The TRAP activity was detected using spectrophotometer, the gene expression of TRAP, receptor activator of NF-κB (RANK), receptor activator of NF-κB lig and (RANKL), and osteoprotegerin (OPG) was measured by RT-PCR, and the RANK protein expression was detected by Western blotting. Results: Compared with the osteoclast group, SPC-I (5 and 10 g/L) groups could make TRAP positive cells and TRAP activity decreased, TRAP, RANK, and RANKL expression in gene level reduced, and RANK expression in protein level down-regulated also (P<0.01); Compared with the control group, SPC-I (2.5 and 10 g/L) could make the OPG expression in gene level increased and the RANKL/OPG ratio declined (P<0.01). The effect of 5 g/L SPC-I was the most significant (P<0.01). The effect of 2.5g/L SPC-I was not significant. Conclusion: SPC-I has the inhibitory effect on the osteoclast formation and differentiation; The effect of implementation is through RANKL/OPG signal transduction pathway to regulate the expression of TRAP and RANK genes.