| Objective: To explore the mechanism of Drynaria Rhizome on invigorating the kidney and building up bones on different levels. To identify the optimum conditions of the SD rat BMSCs cultured with the active component Naringin of Drynaria Rhizome in vitro. To observe the effects on proliferation, differentiation to osteoblast of BMSCs cultured by various concentrations of Naringinn and confirm the best concentrations and duration of action.To investigate the expression of BMP-2 and Cbfal mRNA in induced system with Naringin on molecular biology level. Methods:1. BMSCs from SD rats were isolated and purified by using different attachment method, then identified according to morphology and induced-differentiation potentiality. So we can establish a stable culture system in viro;2. Different concentrations of Naringin were added to BMSCs with high, medium and low concentrations. Proliferation between Naringin groups (different concentrations) and blank control group (standard medium) were detected by means of CFU-F and MTT( CFU-F: 2μg/ml～500μg/ml; MTT: 0.06μg/ml～500μg/ml);3. ALP-positive cell counts by the improvement Gomori's calcium-cobalt staining was explored to observe the osteoblastic differentiation potentiality of BMSCs induced by different-concentration Naringin (2μg/ml～500μg/ml). Blank control is standard medium without Naringin and positive control is the same one with dexamethasone, sodium β-glycerophosphate and vitamine C. Through these we could confirm the most effective concentrations and duration of action with Naringin(2μg/ml, 15 days);4. 4 groups were divided: Naringin group ( induced by the most effective concentrations), blank control, classic group (classic induce bone formation group, positive control), Naringin +classic group; The ALP activity and the amount of osteocalcin in cells were measured to indicate the osteoblast function of different groups;... |
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