Pro-differentiative, Pro-adhesive and Pro-migratory Activities of Isorhamnetin in MC3T3-E1 Osteoblasts via Activation of ERK-dependent BMP2-Smad Signaling.

ABSTRACT

Osteoporosis (OP) is an epidemic bone remodeling disorder of growing relevance with the aging population. Considering that isorhamnetin (ISO), a flavonoid derived from plant, has been newly reckoned as an active ingredient in treating OP, our paper was conducted to investigate the regulatory role and mechanism of ISO in OP. CCK-8 method detected cell activity. Alkaline phosphatase (ALP) assay kit, ALP staining and alizarin red S staining measured osteogenic differentiation. RT-qPCR and Western blot examined the expressions of osteoblast-related proteins. Wound healing and cell adhesion assays severally detected cell migration and adhesion. Also, Western blot tested the expressions of extracellular signal-regulated kinase (ERK) signaling-associated proteins. As illustrated, after MC3T3-E1 pre-osteoblasts were stimulated to differentiate to osteoblasts, ISO markedly promoted the differentiation, mineralization, migration and adhesion of MC3T3-E1 osteoblasts in a concentration-dependent manner. In addition, administration of ISO functioned as an activator of ERK-dependent BMP2-Smad signaling in MC3T3-E1 osteoblasts and pretreatment with ERK inhibitor PD98059 partially compensated the impacts of ISO on MC3T3-E1 osteoblasts differentiation, mineralization, migration as well as adhesion. To be summarized, ISO might activate ERK-dependent BMP2-Smad signaling to facilitate the differentiation, mineralization, migration and adhesion of MC3T3-E1 osteoblasts, suggesting the protective potential of ISO in OP.