左归丸含药血清调节Runx2激活ALP、OPN蛋白促进BMSCs增殖和分化的研究
Study on the promotion of proliferation and differentiation of BMSCs by Runx2 activation of ALP and OPN proteins with Zuoguiwan containing serum
  
DOI:10.3969/j.issn.1006-7108.2022.05.006
中文关键词:  左归丸  Runt相关转录因子2  骨髓间充质干细胞  骨质疏松
英文关键词:Zuoguiwan  runt-related transcription factor 2  bone marrow mesenchymal stem cells  osteoporosis
基金项目:国家自然科学基金项目(81573874,81873229);江苏高校中西医临床医学品牌专业建设工程资助项目(二期)
作者单位
刘飞祥 樊巧玲* 谭峰* 李星 麦聪英 叶素敏 张明玥 柴毅 吴丹 南京中医药大学中医学院·中西医结合学院江苏 南京210023 
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中文摘要:
      目的 探讨左归丸(Zuoguiwan,ZGW)的补肾填精、益髓壮骨的作用,促进骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的增殖和向成骨细胞方向分化。方法 制备ZGW含药血清,设置10% FBS DMEM组、成骨诱导液组、2.5%空白大鼠血清-成骨诱导液组、2.5% ZGW含药血清-成骨诱导液组。采用MTT法观察各组对BMSCs的增殖,使用碱性磷酸酶钙钴染色法检测BMSCs向成骨细胞方向的分化能力,碱性磷酸酶(alkaline phosphatase,AKP)法检测细胞培养液中AKP的含量,Western blot检测BMSCs中Runt相关转录因子2(Runx2)、骨钙蛋白(osteocalcin,OPN)和碱性磷酸酶(alkaline phosphatase,ALP)蛋白的表达。结果 2.5% ZGW含药血清-成骨诱导液组能够促进BMSCs的快速增殖,较早地进入平台期,促进BMSCs中AKP的分泌,提高ALP含量,上调BMSCs细胞中Runx2、ALP和 OPN的蛋白表达水平。结论 ZGW抗骨质疏松的分子机制,可能与调节Runx2激活ALP、OPN蛋白表达,促进BMSCs增殖和向成骨细胞方向分化有关。
英文摘要:
      Objective To study the effects of Zuoguiwan (ZGW) on nourishing kidney essence, strengthening bone and marrow, and promotion of the proliferation of bone marrow mesenchymal stem cells (BMSCs) and differentiation into osteoblasts. Methods The ZGW containing serum was prepared. 10% FBS DMEM group, osteogenic liquid group, 2.5% serum of health rats with osteogenic liquid group, and 2.5% serum containing ZGW osteogenic liquid group were set up. MTT method was used to observe the proliferation of BMSCs in each group. The differentiation of BMSCs into osteoblasts was tested using alkaline phosphatase calcium cobalt staining method. Alkaline phosphatase (AKP) method was used to detect the content of AKP in cell culture medium. The expression of runt related transcription factor 2 (Runx2), osteocalcin (OPN), and alkaline phosphatase (ALP) protein in BMSCs were detected using Western blotting. Results Rapid proliferation of BMSCs was showed in 2.5% serum containing ZGW osteogenic liquid group. The plateau stage appeared earlier. It promoted the secretion of AKP in BMSCs, increased the content of ALP, and up-regulated the protein expression levels of Runx2, ALP, and OPN in BMSCs cells. Conclusion The molecular mechanism of ZGW against osteoporosis may be related to the regulation of Runx2, activation of ALP and OPN protein expression, promotion of BMSCs proliferation and differentiation into osteoblasts.
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