| The development of new medicine for promoting bone repair has been one of the focuses of biomedical research. Therefore, it is of great significance to develop new drugs from the traditional Chinese medicinal materials with limited side effect and to study the mechanism. Polygonum amplexicaule var. sinense is the commonly used medicine by Tujia ethnic group as folk medicine to invigorate blood and so on. Especially, Polygonum amplexicaule var. sinense has remarkable curative effect in the treatment of bone fractures. However, what is the effective components? And how they work? these questions are now unkown.In this study, extracts from Polygonum amplexicaule var.sinensewere used as the material, mice osteoblast cultured in vitro was used as cell model , the effect of extract on osteoblast activity in vitro was studied in order to find the active components in the plant and provide more scientific information to understand the mechanism.Chinese traditional medicine Polygonum amplexicaule var. sinense was extracted with methanol, ethyl acetate, respectively. Extracts of methanol part, ethyl acetate part and water part were obtained. Mouse osteoblasts cultured in vitro was treated with each part of extracts of different concentration, and then, the influence of the these parts on the osteoblast growth and osteogenesis activity of osteoblast were studied.Osteoblasts were seperated from the calvaria of Kuming mice of 2-3 days old. Osteoblast was cultured in DMEM with 10% of newborn calf serum. All the extracts from Polygonum amplexicaule var. sinense was diluted to a final concentration of 10-2, 10-3, 10-4, 10-5mg/ml and their effect on the osteoblast proliferation, cell cycle and the alkaline phosphatase activity were studied. CCK-8 method was used to study the influence of extracts from Polygonum amplexicaule var. sinense on activity of the proliferation osteoblast. Result showed that methanol extracts, ethyl acetate extracts, water extracts with the concentration range of 1.0 x 10-3, 10-4, 10-5mg/ml can promote the proliferation of osteoblast (p<0.05). Flow Cytometry technique was used to study the effect of the extracts on cell cycle. The results showed that the methanol extracts in 1.0 x10-3mg/ml,ethyl acetate extract in 1.0 x10-3,10-4,10-5 mg/ml, water extracts in 1.0 x10-2,10-3,10-4 mg/ml concentration range could promote the osteoblast into the cell cycle S period, which meant that cell proliferation index was enhanced. Higher concentration would inhibit the cells into the cell cycle S period, thus, cell proliferation index was lower.The effect of extract on alkaline phosphatase activity of osteoblasts was studied using alkaline phosphatase kit and the result showed that methanol part in 1.0 x 10-3, 10-4mg/ml, ethyl acetate part in 1.0 x10-3,10-4,10-5 mg/ml, and water parts in 1.0 x 10-3mg/ml of Polygonum amplexicaule var. sinense could promote the activity of alkaline phosphatase in osteoblasts.Conclusion: to sum up, methanol extracts, ethyl acetate extracts, water extracts from Polygonum amplexicaule var. sinense can promote the proliferation and differentiation of osteoblast. |
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