叶亚平，李娜，向威，王锐，郭风劲，许涛.周期性张应力联合淫羊藿苷促进脂肪干细胞成骨分化的实验研究[J].中华物理医学与康复杂志,2017,39(1):5-10
扫码阅读全文
|
| 周期性张应力联合淫羊藿苷促进脂肪干细胞成骨分化的实验研究 |
|
| |
| DOI： |
| 中文关键词: 淫羊藿苷 周期性张应力 脂肪干细胞 成骨分化 |
| 英文关键词: Icariin Cyclic tensile strain Adipose-derived stem cells Osteogenic differentiation |
| 基金项目:国家自然科学基金面上项目（81371915） |
|
| 摘要点击次数: 2480 |
| 全文下载次数: 3308 |
| 中文摘要: |
| 目的探讨周期性张应力（CTS）联合淫羊藿苷促进脂肪干细胞（ASCs）成骨分化的作用。 方法分离10只Sprague-Dawley(SD)大鼠的ASCs，分别按不同的干预方法将收集的ASCs细胞分为对照组（仅用普通培养基干预）、淫羊藿苷组（仅用淫羊藿苷干预）、1000μ组（仅用1000μ的CTS干预）、2000μ组（仅用2000μ的CTS干预）、3000μ组（仅用3000μ的CTS干预）和联合干预组（用2000μ的CTS联合淫羊藿苷进行干预）。淫羊藿苷的作用浓度为10-7mol/L；CTS的设置参数为频率1.0Hz，应力大小分别为1000μ strain（简称1000μ）、2000μ、3000μ，每刺激5s之后休息15s，每日总刺激时间为2h。干预7d后,收获细胞采用Western Blot法检测碱性磷酸酶（ALP）蛋白的表达；用淫羊藿苷联合2000μ的CTS对ASCs进行干预3d后，收获ASCs细胞用Western Blot法检测其Runt相关基因转录因子2（Runx2）、YES相关蛋白（YAP）和结缔组织生长因子（CTGF）蛋白的表达；分别于干预3d和7d后，采用ALP活性定量检测试剂盒对ALP活性进行检测；干预3d后，利用RT-PCR检测YAP下游靶基因CTGF和锚蛋白重复结构域1（Ankrd1）的表达；干预7d后，利用RT-PCR检测成骨分化相关基因骨桥素（OPN）和Ⅰ型胶原的表达，并对各组所得数据进行统计学分析比较。 结果淫羊藿苷和CTS（1000μ、2000μ、3000μ）均能显著促进ALP蛋白的表达。与1000μ和3000μ应力大小的CTS相比，2000μ的CTS具有最强的促ALP蛋白表达能力；且2000μ的CTS联合淫羊藿苷能显著促进ALP的表达及Runx2和CTGF的表达，且其联合作用时效应更明显；但淫羊藿苷和2000μ的CTS单独作用并不能促进YAP蛋白的表达，只有联合作用时才具有促YAP表达作用。淫羊藿苷和2000μ的CTS单独作用3d和7d后均能显著上调ALP的活性，且其联合作用时上调ALP活性的效应更加明显。淫羊藿苷和2000μ的CTS单独作用7d后，能显著促进成骨分化基因OPN和Ⅰ型胶原蛋白的表达，且联合作用时效应更明显。淫羊藿苷和2000μ的CTS单独作用3d后能显著促进YAP靶基因CTGF和Ankrd1的表达，联合作用时效应更明显。 结论CTS和淫羊藿苷联合作用可以通过上调YAP活性来促进ASCs的成骨分化。 |
| 英文摘要: |
| Objective To study the roles of icariin and cyclic tensile strain (CTS) in promoting the osteogenic differentiation of adipose-derived stem cells (ASCs) and the molecular mechanisms involved. MethodsASCs were isolated from Sprague-Dawley rats and treated either with icariin (10-7mol/L) or with 1000μ, 2000μ or 3000μ of CTS for 7 days, or with icariin plus CTS at 2000μ for three days. Alkaline phosphatase (ALP) activity was detected after 3 and 7 days of intervention. Western blotting was performed to detect the expression of Runt-related transcriptional factor 2 (Runx2), Yes-associated protein (YAP) and connective tissue growth factor (CTGF) after the third day of the intervention. A reverse transcription polymerase chain reaction was performed at 7 days to detect the expression of osteopontin (OPN) and collagen 1a and after 3 days to detect the expression of the YAP target gene, CTGF and ankyrin repeating domain 1 (Ankrd1). ResultsIcariin and CTS at 1000μ, 2000μ or 3000μ could all significantly promote the expression of ALP protein. CTS at 2000μ was the most effective. The co-treatment with icariin and CTS significantly promoted ALP protein expression compared with icariin or CTS treatment alone. It also significantly promoted the expression of Runx2 and CTGF protein. Icariin or CTS (2000μ) alone could not promote the expression of YAP protein, but icariin combined with CTS (2000μ) promoted it significantly. Either icariin or CTS (2000μ) could significantly promote ALP activity after 3 and 7 days, but icariin combined with CTS had the most obvious effect. Both icariin and CTS (2000μ) could also significantly promote the expression of the osteogenesis-related genes OPN and collagen 1a, as well as the YAP targeted genes CTGF and Ankrd1. However, the combination of icariin and CTS had the greatest effect in promoting the expression of OPN mRNA, collagen 1a mRNA, CTGF mRNA and ankrd1 mRNA. ConclusionIcariin and CTS co-treatment may promote osteogenic differentiation of ASCs via activating YAP expression. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
