摘要
目的从细胞生物学角度观察葛根素对犬骨髓基质细胞的成骨诱导作用,探讨葛根素的促成骨效果。方法原代培养犬骨髓基质细胞,第三代时加入葛根素,实验分四组:实验组3组DMEM培养基+葛根素(葛根素液浓度分别为0.01、0.1、1mmol/L);对照组1组为DMEM培养基(无葛根素)。用倒置显微镜观察细胞形态,CCK-8检测药物毒性,茜素红染色观察钙结节,最后用SPSS20.0软件对数据行统计分析。结果葛根素液诱导细胞后,能明显促进细胞增殖,且葛根素浓度为0.1 mmol/L时促增殖最明显,与空白对照组比较差异有显著性(P<0.05),药物诱导后各组碱性磷酸酶活性增加,其中药物浓度为0.1 mmol/L的实验组与另两组比较有明显差异(P<0.05),葛根素诱导培养16 d后可观察到钙结节形成。结论葛根提取物葛根素能明显促进犬骨髓基质细胞的增殖和成骨分化,其中浓度为0.1 mmol/L时效果最好。
Objective This paper observes the induced function of puerarin on the canine bone marrow stromal cells from a biological point and investigates the osteogenic effect of puerarin on the bone. Methods This experiment is divided into 4 groups. At first, we culture some canine bone marrow stromal cells. And then during the third generation, add some of the puerarin. The experimental group 3 ： DMEM medium ＋ puerarin （ puerarin night concentra- tions were 0.01mmol / 1,0. lmmol / 1, lmmol / 1） ,the control group（ the first group）DMEM medium（ no puerarin）. After that, we invert the microscope to observe the cell morphology, CCK - 8 detection of drug toxicity ; measuring the activity of alkaline calcium nodules ; Alizarin red staining of calcium nodules and finally we use SPSS20.0 statistical software to analyze data. Results After the induction of cell puerarin solution, we found that it can significantly promote cell proliferation and when the concentration of puerarin is 0.1 mmol/1, the proliferation is the most obvious. Whereas in the case of the control group, there was a significant difference （ P 〈 0.05 ）. The activity of Alkaline phosphatase increased in each group after the induction of the drug, among which the concentration of drug is 0.1 mmol / 1 is significantly different in the experimental group by comparison with the control group （P 〈 0.05 ）. After induction culture of 16d puerarin ,we can observe the formation of calcium nodule. Conclusion The Kudzu extract from puerarin can significantly promote the proliferation of canine bone marrow stromal cells and osteoblasts differentiation, and the best effect takes place wherein the concentration is 0.1 mmol / 1.
出处
《青海医学院学报》
CAS
2014年第2期123-127,共5页
Journal of Qinghai Medical College
关键词
葛根素
犬骨髓基质细胞
成骨诱导
Puerarin Canine bone marrow stromal cells Osteogenic
