Osteoblasts Proliferation and Differentiation Stimulating Activities of the Main Components of Epimedii folium

BACKGROUND Osteoporosis is a disease of bones that leads to an increased risk of fracture. Epimedii Folium is commonly used for treating bone fractures and joint diseases for thousands of years in China. METHODS This study was aimed to screen active components, which might have the potency to stimulate osteoblasts proliferation and differentiation in Epimedii Folium. An HPLC method was established to analyze the main components in Epimedii Folium. The MTT and ALP methods were utilized for the assay of osteoblasts proliferation and differentiation activity. Bavachin, a flavonoid compound was treated as the positive control. RESULTS Totally eight compounds have been identified by comparing their retention time with correspondent standard substances. Icariside I and icariside II significantly stimulated cell proliferation and osteoblasts differentiation. All these compounds were found with a characterized flavonoid structure in each of their molecule backbones. CONCLUSION These results lead to a hypothesis that flavonoid monoglycoside structure might be crucial to exhibit the activity. The structure-effect relationship of these compounds with flavonoid monoglycoside structure in mouse primary calvarial osteoblasts needs to be explored in further research. SUMMARY Eight compounds were identified by comparing their retention time with correspondent standard substances.Icariside I and icariside II significantly stimulated cell proliferation and osteoblasts differentiation.Flavonoid monoglycoside structure might be crucial to exhibit the osteoblasts proliferation and differentiation activity. Effects of the main components of Epimedii Folium on osteoblasts proliferation after treating 48 h. Abbreviations used: HPLC: High performance liquid chromatography, MTT: Methylthiazolyldiphenyl - tetrazolium bromide, ALP: Alkaline phosphatase.