吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (03): 491-495.doi: 10.13481/j.1671-587x.20150311

• 基础研究 • 上一篇    下一篇

鹿茸多肽对人骨髓间充质干细胞BMP-2和Runx2表达的影响

张洪长1, 张莹2, 刘明昕3, 潘志1, 律广富1, 陈港1, 姜鸿远1, 郭骏骐4   

  1. 1. 长春中医药大学中医药与生物工程研究开发中心药理研究室, 吉林 长春 130117;
    2. 吉林大学药学院药理学教研室, 吉林 长春 130021;
    3. 长春中医药大学图书馆参考咨询部, 吉林 长春 130117;
    4. 长春中医药大学科技处, 吉林 长春 130117
  • 收稿日期:2014-10-09 发布日期:2015-08-01
  • 通讯作者: 郭骏骐,副研究员(Tel:0431-86045421,E-mail:kjcchgk@163.com) E-mail:kjcchgk@163.com
  • 作者简介:张洪长(1961-),男,吉林省长春市人,副研究员,医学博士,主要从事分子生物化学与药理学方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(81374025);吉林省科技厅科技发展计划项目资助课题(20140414051GH)

Influence of velvet antler polypeptides in expressions of BMP-2 and Runx2 in human bone marrow mesenchymal stem cells

ZHANG Hongchang1, ZHANG Ying2, LIU Mingxin3, PAN Zhi1, LYU Guangfu1, CHEN Gang1, JIANG Hongyuan1, GUO Junqi4   

  1. 1. Department of Pharmacology, Center of Traditional Chinese Medicine and Bioengineering, Changchun University of Chinese Medicine, Changchun 130117, China;
    2. Department of Pharmacology, School of Pharmacy, Jilin University, Changchun 130021, China;
    3. Department of Reference, Library of Changchun University of Chinese Medicine, Changchun 130117, China;
    4. Department of Technology, Changchun University of Chinese Medicine, Changchun 130117, China
  • Received:2014-10-09 Published:2015-08-01

摘要:

目的:探讨鹿茸多肽(VAP)对人骨髓间充质干细胞(hBMSCs)中骨形态发生蛋白2(BMP-2) 和骨特异性转录因子2(Runx2)基因和蛋白表达的影响,阐明其对骨合成及hBMSCs向成骨细胞方向分化的作用机制。方法:体外培养的hBMSCs分为对照组和5、10、15、20 g·L-1 VAP组,通过检测各组hBMSCs中碱性磷酸酶(ALP)活性,确定体外用药最佳药物浓度;CCK-8法检测对照组和10 g·L-1 VAP组hBMSCs增殖情况;荧光定量 PCR 法与Western blotting法测定对照组和10 g·L-1 VAP组hBMSCs中BMP-2和Runx2基因与蛋白的表达。结果:VAP各组细胞中ALP活性均高于对照组,其中以10 g·L-1 VAP组细胞中ALP活性最高(P<0.01);CCK-8法,10 g·L-1 VAP组hBMSCs增殖率明显高于对照组(P<0.05),VAP诱导培养第9天hBMSCs增殖率最高,且进入平台期,之后hBMSCs的增殖率开始下降;荧光定量 PCR和Western blotting检测,10 g·L-1 VAP组hBMSCs中BMP-2和Runx2基因及蛋白表达水平明显高于对照组,差异有统计学意义(P<0.05)。结论:VAP能够提高hBMSCs中ALP活性,促进hBMSCs增殖分化,上调hBMSCs中BMP-2及其下游Runx2基因和蛋白的表达,这可能是VAP对hBMSCs骨形成及骨代谢影响的分子调控机制之一。

关键词: 鹿茸多肽, 人骨髓间充质干细胞, 骨形态发生蛋白2, 骨特异性转录因子2

Abstract:

Objective To investigate the influence of velvet antler polypeptides (VAP) in the expressions of bone morphogenetic protein-2 (BMP-2) and runt-related transcription factor 2 (Runx2) gene and protein in human bone marrow mesenchymalstem cells (hBMSCs), and to clarify the mechanism of VAP in bone synthesis and bone differentiation direction. Methods The cultured hBMSCs were divided into control group and 5, 10, 15, 20 g ·L-1 VAP groups.By detecting the alkaline phosphatase (ALP) activities in hBMSCs in various groups, the optimal concentration of the drug in vitro was selected;CCK-8 assay was used to detect the proliferation of hBMSCs in control group and 10 g·L-1 VAP group;the BMP-2 and Runx2 gene and protein expressions in control group and 10 g·L-1 VAP group were determined with fluorescence quantitative PCR and Western blotting method. Results The ALP activities in VAP groups were higher than that in control group, and the ALP activity in 10 g ·L-1 VAP group was the highest (P<0.01).The CCK-8 assay Results showed that the proliferation rate of the hBMSCs in 10 g ·L-1 VAP group was higher than that in control group (P<0.05), the VAP-induced proliferation of cultured hBMSCs was the highest at the 9th day, then went into the plateau and began to decline;the quantitative PCR and Western blotting Results showed that the expression levels BMP-2 and Runx2 gene and protein in the hBMSCs in 10 g ·L-1 VAP group were significantly higher than those in control group, and the differences were statistically significant(P<0.05). Conclusion VAP can improve the ALP activity in hBMSCs and promote the proliferation and differentiation of hBMSCs, and increase the expressions of BMP-2 gene and its Runx2 gene and protein, which may be one of the molecular regulation mechanisms of VAP for bone formation and bone metabolism of hBMSCs.

Key words: velvet antler polypeptides, human bone marrow mesenchymal stem cells, bone morphogenetic protein-2, runt-related transcription factor 2

中图分类号: 

  • R961