Effects Of Plateau Propolis On The Proliferation And Differentiation Of Osteoblast In Vitro

[Objective]To study the diffusion method of plateau propolis and its effect on the proliferation and differentiation of osteoblast in vitro.[Materials and Methods]1.The raw extraction of propolis were prepared with 95% ethanol,and two experimental groups,hydrosoluble component group and alcohol-soluble component group were extracted by reversed phase high performance liquid chromatogram and diphase solvent extraction.The flavanoid content of propolis extration were detected by ultraviolet spectrophotometry(UV spectrophotometry).2.Osteoblast derived from fetal rat calvarial were cultured on 24-well and 96-well plates with propolis extraction of two experimental groups in the concentration of 180μg/ml,150μg/ml,120μg/ml,90μg/ml,60μg/ml and 30μg/ml respectively.The effects of hydrosoluble component group and alcohol-soluble component group on the proliferation and differentiation of osteoblast were measured by MTT assay,AgNOR,PNPP,CBB and FCM.[Results]1.The UV spectrophotometry results showed the content of flavanoid in hydrosoluble component group and alcohol-soluble component group were 22.73% and 25.27%respectively.2.From the results of MTT assay,the OD value increased in alcohol-soluble component group,while decreased in hydrosoluble component group.AgNOR analysis results showed two experimental groups had no effect on the DNA\RNA synthesis,abnormal proliferation and chromosome aberration of osteoblasts the alcohol-soluble group showed inhibitory effect on the ALP synsthesis of osteoblasts,while hydrosoluble component group had no such effect.The 150μg / ml,120μg / ml,90μg / ml,60μg / ml and 30μg /ml of alcohol-soluble component group showed significant effect on the differentiation and protein synthesis of osteoblasts from the results of CBB test.There is statistic difference between 60μg/ml of alcohol-soluble component group and control group.The FCM results indicated that 90μg/ml,60μg/ml and 30μg/ml of hydrosoluble component group and 30μg/ml of alcohol-soluble component group could prompt osteoblasts from stage Go(G₁)to stage S,G₂.[Conclusions]1.The content of flavanoid in hydrosoluble component group and alcohol-soluble component group were 22.73%and 25.27%respectively.2. Propolis extraction showed two-ways regulation effect on the proliferation and differentiation of osteoblast.3.AgNOR analysis results proved that propolis extraction did not result in chromosome aberration and had no cytotoxicity.4. Alcohol-soluble component of propolis could reduce the cytotoxic effect of ethanal on osteoblast.