黄芪多糖对 MC3T3-E1 细胞增殖及成骨分化的影响.

Effects of Astragalus polysaccharides on the proliferation and osteogenic differentiation of MC3T3-E1 cells.

Objective To study the effects and mechanisms of Astragalus Polysaccharides (AP) on the proliferation and osteogenic differentiation of MC3T3-E1 cell line. Methods MC3T3-E1 cells were cultured in different concentrations of AP (0, 5, 10μmol/L) for the intervention. The cell viability was detected by MTT method on the first, third, fifth, and seventh day; and cells were stained with ALP and Alizarin Red, and real-time PCR were used to detect the mRNA levels of osteogenic differentiation-related markers, including Runt-related transcription factor 2 (Runx2), β-catenin (β-catenin), and osteocalcin (Osteocalcin), Collagen I (Collagen I); Western blot was used to detect the expression levels of β-catenin, Osteocalcin, and Runx2 proteins, and immunofluorescence was used to detect the nuclear translocation of β-catenin. Results The addition of 5 and 10μM AP could effectively promote the proliferation of MC3T3-E1; at the same time, it could also increase the positive rate of ALP and Alizarin Red staining, as well as β-catenin and Osteocalcin, Runx2, Collagen I mRNA expression levels (P<0.05); Western blot results also showed that AP can effectively promote the expression of β-catenin, Osteocalcin, Runx2 protein (P<0.05), and promote β-catenin into the nucleus. Conclusion AP can promote the proliferation and osteogenic differentiation of MC3T3-E1. The potential mechanism may be related to the up-regulation of osteogenic markers and the promotion of β-catenin into the nucleus. [ABSTRACT FROM AUTHOR]

目的 分析黄芪多糖（AP）对小鼠成骨细胞系MC3T3-E1的成骨分化作用机制。 方法 MC3T3-E1细胞经成骨诱导后分别加入浓度为0、5、10 μmol/L 的AP进行干预，持续1、3、5、7 d后使用MTT法观察AP对细胞增殖的影响；干预14 d后，通过ALP及茜素红染色观察AP的促成骨分化作用；成骨相关mRNA表达水平经实时荧光定量PCR测定，其中包括Runt相关转录因子2(Runx2)、β-连环蛋白(β-catenin)、骨钙素（osteocalcin）、I型胶原蛋白（collagen I）；并使用Western blot法检测β-catenin、osteocalcin、Runx2蛋白表达水平，以及免疫荧光检测β-catenin的核易位情况。 结果 加入5、10 μmol/L 的AP后可有效促进MC3T3-E1增殖，且药物浓度越高效果越明显；同时AP亦可提高ALP、茜素红染色的阳性率，以及β-catenin、osteocalcin、Runx2、collagen I mRNA的表达水平（P<0.05）；Western blot结果也显示AP可有效促进β-catenin、osteocalcin、Runx2蛋白表达（P<0.05），以及促进β-catenin入核。 结论 AP可促进MC3T3-E1细胞增殖及成骨分化，机制可能涉及促进相关成骨标志物表达以及β-catenin入核。 [ABSTRACT FROM AUTHOR]

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