Effect of extract from callus of Saussurea involucrate on proliferation and differentiation of osteosarcoma cell MG-63 / 中草药

ABSTRACT

Objective: To investigate the effect of extract from callus of Saussurea involucrate (ESI) on the proliferation and differentiation of osteosarcoma cells MG-63 and explore its anti-osteoporosis and mechanism. Methods: Using human osteosarcoma cells MG-63 for the study, the effect of ESI on the proliferation was detected by MTT and LDH methods; The activity of alkaline phosphatase activity (ALP) and the level of hydroxyproline (Hyp) were investigated by related reagent kit; The effect of ESI on mineralized nodules was observed by Alizarin red staining and quantified by CPC, the expression of OPG/RANKL was measured by RT-PCR and Western blotting, and the proliferation, differentiation, mineralization, and OPG/RANKL expression levels of MG-63 were assayed when medium contains both p38 inhibitor SB203580 and ESI. Results: MTT and LDH assay showed that ESI (< 125 μg/mL) was non-toxic to MG-63, and it promoted the proliferation at the concentration of 31.25 and 62.5 μg/mL; ESI could significantly increase the ALP activity, Hyp content, and amount of mineralized nodules; At the mRNA level, ESI could significantly up-regulate OPG expression and down-regulate RANKL expression; At the protein level, ESI could significantly increase the ratio of OPG/RANKL; SB203580 could reverse the acceleration of ESI on the proliferation, differentiation, mineralization, and OPG/RANKL expression of MG-63. Conclusion: ESI has the facilitating effect on the osteoblast proliferation, differentiation, and mineralization; The mechanism may be associated with the expression of OPG and RANKL, as well as the signal transduction pathway of p38 Mitogen-activated potein kinase (MAPK).