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
  
DOI:10.3969/j.issn.1006-7108.2021.12.012
中文关键词:  黄芪多糖  成骨分化  骨质疏松
英文关键词:astragalus polysaccharides  osteogenic differentiation  osteoporosis
基金项目:国家自然科学基金(81860151);江西省重点研发计划(20192BBG70027)
作者单位
胡俊1 杨盼盼1 袁忠1 谈荣珍1 张楚焌1 张斌1 沈云峰2* 1.南昌市洪都中医院骨伤七科,江西 南昌 330008 2.南昌大学第二附属医院内分泌代谢科,江西 南昌 330006 
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中文摘要:
      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。
英文摘要:
      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.
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