Effect of Eucommia Ulmoids Leaf on SD rat osteoblast proliferation and expression levels of osteocalcin

Objective To study different concentrations of Eucommia extract effect on osteoblast proliferation and osteocalcin expression. Methods Newborn SD rats( 24 ~ 48 h,6) are isolated by enzymatic digestion to get primary neonatal rat osteoblasts,osteoblasts were identified by alkaline phosphatase staining; First each groups were added with serum-free medium without phenol red starved SD rats osteoblasts at 2h,then give five groups concentrations Eucommia extract(0mg/ml,1mg/ml,10 mg/ml,50 mg/ml,100 mg/ml) interfere in SD rat osteoblasts and cells were incubated for 48 hours,set the control group(0mg/ml),then MTT method detect cell proliferation; ibid steps,Western blot detect the expression levels of osteocalcin. Results After alkaline phosphatase staining,SD rats osteoblasts was purple,MTT method showed that the leaf extract on the proliferation of osteoblasts have a role in promoting(P 0. 05) Compared with the control group,proliferation of osteoblas tsassociated with the drug concentration,wherein the most significant concentration was 50 mg/ml(P 0. 01). Western bolt results are shown that each dosing groups of osteocalcin protein levels were significantly elevated(P 0. 05) compared with the control group,and also has a consistent concentration-dependent. Conclusion Eucommia extract can promote osteoblast proliferation through affecting the expression of osteocalcin and osteoblast proliferation has a concentration-dependent nature.