Fig. 6. Effect of P. japonicus leaf extract on Runx2 and Osterix mRNA expressions in MC3T3-E1 cells.

MC3T3-E1 cells were plated at 2 × 10⁴ cells/well in 48-well plates and incubated for 24 h. After incubation for 24 h, cells were incubated for 3 days in ObDIM containing various concentrations of AL and E2. Total RNA in cells was extracted, reverse transcribed, and real-time PCR was conducted. Relative mRNA expression levels of Runx2 and Osterix in MC3T3-E1 cells were analyzed. Targeted mRNA expression was normalized to that of GAPDH and is presented relative to the ObDIM-treated group. Each bar represents the mean ± standard error of the mean (n = 4).

Runx2, Runt-related transcriptional factor 2; ObDIM, osteoblast differentiation-inducing medium; AL, the aqueous extract of P. japonicus leaves; E2, 100 nM 17-estradiol; PCR, polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

^***P < 0.001 significantly different from the ObDIM-untreated group. Means without a common letter differ significantly at P < 0.05.