论著·基础研究

小分子抗菌肽 KR -12-a5促进人骨髓间充质干细胞成骨分化的作用

  • 李晖 * ,
  • 张术涛 * ,
  • 岳冰
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  • 上海交通大学医学院附属仁济医院骨关节外科,上海 200011
李晖(1993—),男,硕士生;电子信箱: jiayou1101@163.com。张术涛(1993—),男,硕士生;电子信箱: shutao.zhang@me.com。*为共同第一作者。

网络出版日期: 2018-09-11

基金资助

国家自然科学基金( 81472119,81672196);上海市教育委员会高峰高原学科建设计划( 20161432)

Small peptide KR-12-a5 promotes the osteogenic differentiation of human bone marrow mesenchymal stem cells

  • LI Hui* ,
  • ZHANG Shu-tao* ,
  • YUE Bing
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  • Department of Bone and Joint Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China

Online published: 2018-09-11

Supported by

National Natural Science Foundation of China, 81472119, 81672196; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Support, 20161432

摘要

目的 ·验证抗菌肽 KR-12的类似物 KR-12-a5对人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,HBMSCs)成骨分化功能的影响。方法 ·使用 HBMSCs进行成骨分化相关实验,使用 KR-12-a5作为成骨诱导条件下的附加刺激,并通过碱性磷酸酶(alkaline phosphatase,ALP)染色实验、ALP定量实验、茜素红染色和洗脱定量分析以及成骨相关基因实时荧光定量 PCR检测,验证 KR-12-a5对 HBMSCs成骨分化功能的影响。结果 ·随着 KR-12-a5刺激浓度的增加, ALP和茜素红的染色强度也随之增加;在最高浓度( 50 μg/mL)的 KR-12-a5刺激下显示了最强的染色效果,定量测试也显示出类似的结果。在 KR-12-a5刺激 3 d后,RUNX2的 mRNA水平以药物浓度依赖的方式增加;在第 7日,KR-12-a5处理组中的 ALP、COL1A1、BSP和 BMP2的表达水平表现出明显的变化;到第 14日,OSX、OCN和 OPN水平显著增加。结论 · KR-12-a5能以药物浓度依赖的方式促进 HBMSCs的成骨分化功能。

本文引用格式

李晖 * , 张术涛 * , 岳冰 . 小分子抗菌肽 KR -12-a5促进人骨髓间充质干细胞成骨分化的作用[J]. 上海交通大学学报(医学版), 2018 , 38(8) : 881 . DOI: 10.3969/j.issn.1674-8115.2018.08.005

Abstract

Objective · To investigate the effect of KR-12 analog, KR-12-a5, on osteogenic differentiation of human bone marrow mesenchymal stem cells (HBMSCs). Methods · Osteogenic differentiation-related experiments were performed using HBMSCs. KR-12-a5 was used as additional stimulation under osteogenesis inducing environment.alkaline phosphatase (ALP) staining assay, ALP quantitative assay, alizarin red staining and elution quantitative analysis, and real-time quantitative polymerase chain reaction (RT-qPCR), the effect of KR-12-a5 on osteogenic differentiation of HBMSCs was verified. Results · As the concentration of KR-12-a5 increased, the staining intensity of ALP and alizarin red also increased. The strongest staining effect was exhibited at the highest concentration (50 μg/mL) of KR-12-a5. Quantitative tests showed similar results. After 3 days of stimulation with KR-12-a5, the mRNA level of RUNX2 increased in a dose-dependent manner. On the 7th day, the levels of ALP, COL1A1, BSP and BMP2 in the KR-12-a5-treated group showed significant changes. On the 14th day, OSX, OCN and OPN levels increased significantly. Conclusion · KR-12-a5 promoted the osteogenic differentiation of HBMSCs in a dose-dependent manner.
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