Bacillus subtilis Fermentation of Malva verticillata Leaves Enhances Antioxidant Activity and Osteoblast Differentiation

Malva verticillata, also known as Chinese mallow, is an herbaceous plant with colorful flowers and has been used as a medicine for thousands of years. This study investigated this herb for potential antioxidant activity or an association with osteoblast differentiation. M. verticillate leaves were fermented with B. subtilis MV1 at 30 °C for 7 days to enhance their biological activities. The resultant aqueous extract (MVW) and the fermented leaves (MVB) were measured for antioxidant and osteoblast differentiation. The results showed that the total phenolic, flavonoid, and antioxidant activity, as well as the osteoblast differentiation of the MVB increased (2 to 6 times) compared with those of the MVW. MVB induced phosphorylation of p38, extracellular signal-regulated kinase in C3H10T1/2 cells, and the phosphorylation was attenuated via transforming growth factor-β (TGF-β) inhibitors. Moreover, runt-related transcription factor 2 and osterix in the nucleus increased in a time-dependent manner. The messenger RNA expression of alkaline phosphatase and bone sialoprotein increased about 9.4- and 65-fold, respectively, compared to the non-treated cells. MVB stimulated C3H10T1/2 cells in the osteoblasts via TGF-β signaling. Thus, fermented M. verticillata extract exhibited enhanced antioxidant activity and osteoblast differentiation.