The Study Of The Effects Of Radix Puerariae On The Expressions Of OPG And RANKL MRNA In Rat Osteoblasts

ObjectiveThis subject, by establishing experimental model of rat's cultured osteoblast in vitro, investigates the effects of Puerarin on the proliferation and differentiation and gene expression of rat osteoblast in cell and molecular level,in order to approach the mechanism of antiosteoporotic effects of Puerarin on postmenopausal osteoporosis and provide the experimental evidences for its preventing and treating postmenopausal osteoporosis in clinic.Methods1.The osteoblasts are isolated from newborn rats within 24 hours by digesting rat's calvaria repeatedly,and collected and cultured in vitro. Morphologic changes of osteoblasts are observed.Rat osteoblasts are identified by the alkaline phosphatase staining2.To observe the effects of Puerarin on the proliferation of rat osteoblasts by using MTT method3.To explore the effects of Puerarin on the expressions of osteoprotegerin (OPG) and receptor activator of nuclear factor-icB ligand(RANKL) by RT-PCR in rat osteoblasts cultured in vitro.Results1.Experimental model of rat's cultured osteoblast in vitro is established.The cells possesses the morphological and biological properties of osteoblast.2.All of the concerntrations are promoted by the effect of Puerarin with different concentrations on the rat osteoblasts for 24 hours and 48 hours;After cultivated 72 hours and 96 hours,all of the concentrations has no effects on proliferation3.Puerarin can increase OPG mRNA levels remarkably and decrease RANKL mRNA levels in the rat osteoblasts cultured in vitro Conclusions1.Puerarin with different concentrations can promote the proliferation of osteoblasts remarkably.and the mount of proliferation has intimate relations with concentration and time2.Puerarin can upregulate OPG expression and downregulate RANKL expression in rat osteoblasts,which may explain the mechanism of antiosteoporotic effects of Puerarin in preventing and treating postmenopausal osteoporosis.3.Puerarin can promote the proliferation of osteoblasts and the expressions of osteoprotegerin (OPG) and receptor activator of nuclear factor-KB ligand(RANKL) mRNA,which may be the mechanism of the effects of Puerarin on postmenopausal osteoporosis.Our study not only provides the experimental evidences for the application of Puerarin,also gives a new clue on the development of the antiosteoporosis drugs.