Preliminary Study Of Chlorogenic Acid On Activity Of Cultured Osteoblasts In Vitro

Sambucus chinensis Lindl belongs to caprifoliaceae and sambucus. Its root, stem andleaf can be used as a medicine. In traditional Chinese medicine, it is often used to treatrheumatism, traumatic injury and fracture. At present，there are many reports of Sambucuschinensis Lindl about preventing osteoporosis and promoting proliferation of osteoblasts,however, the effective component for promoting fracture healing is not clear. The aim of thisstudy is to investigate the influence of chlorogenic acid——one main ingredient of Sambucuschinensis Lindl on osteoblasts cultured in vitro, in order to identify whether Sambucuschinensis Lindl has the effect of promoting osteogenesis and provide theoretical basis forclinical applications of Sambucus chinensis Lindl.The main content of this study is to use osteoblastic cell line MC3T3-E1as the cellmodel in vitro to evaluate the effect of CGA on the proliferation rate, ALP activity andexpression of bone-related genes in osteoblasts with MTT assay, alkaline phosphatase activitytest, real-time PCR technique.The MTT assay showed that11.02~88.19μmol/L of CGA could obviously promotethe proliferation rate of osteoblasts. Thus, we used22.05μmol/L and44.09μmol/L of CGAin the following study.The assay of influence of CGA on the ALP activity of osteoblasts showed that the CGAinhibited the ALP activity of osteoblasts when treated for long time.Real-time PCR revealed the expression of Col-1, OCN and BMP-2were enhancedduring the whole treatment period; the expression of Jun-D, c-Jun, c-fos, Fra-1, Fra-2, ALP,Runx2, and Osterix were promoted when treated for short term (2days). The expression ofBSP, OPN were promoted when treated for long term (6days).In conclusion, CGA has the ability to regulate the proliferation and differentiation ofosteoblasts. It has some inhibitory effect on the expression of genes responsible for earlydifferential stage in osteoblasts in some degree. It is not beneficial for stem cell todifferentiate to osteoblast. However, CGA could promote the maturation of new bone bymeans of accelerating the formation of bone matrix and bone maturation. Therefore, CGA has osteogenic ability, it is one of the effective components inSambucus chinensis Lindl.