To determine whether Ursolic acid and Oleanolic acid could be used as dietary supplements for osteoporosis animals, CCK-8 (cell counting kit -8), ELISA (enzyme-linked immunosorbent assay), FQ-PCR (Real-time Fluorescence Quantitative PCR) and western bolt assays were used on MC3T3-E1 cells. After treatment with Ursolic acid and Oleanolic acid for 48h and 72h, the cell viability showed marked increased. Ursolic acid and Oleanolic acid could up-regulated RANKL protein in supernatant at 48h, and up-regulated both OPG and RANKL protein at 72h, in different pattern. Ursolic acid increased RANKL mRNA at 48h and 72h, OPG mRNA at 72h; Oleanolic acid increased OPG mRNA at 48h and 72h, RANKL mRNA at 72h. Adapt concentrations of Ursolic acid and Oleanolic acid could up-regulated OPG and down-regulated RANKL protein expression. After exposed on Ursolic acid or Oleanolic acid with or without inhibitors for 72h, PD98059, SB203580, SP600125 and LY-294002 could affect Ursolic acid- and Oleanolic acid-induced cell proliferation and the leakage of OPG proteins. Ursolic acid and Oleanolic acid could regulate the phosphorylation of ERK, JNK, p38 and AKT. So Ursolic acid and Oleanolic acid could promote MC3T3-E1 cells proliferation, OPG and RANKL expression dose-dependently and time-dependently, and Ursolic acid had larger range of concentrations and better effect. These osteogenic effects of Ursolic acid and Oleanolic acid related to MAPK and AKT signaling pathways by different patterns.

Key words: MC3T3-E1, Oleanolic acid, OPG, Osteoporosis, RANKL, Ursolic acid