贺燕婷#,李振凤#,崔星慧,等.岩藻黄素在振荡剪切应力致EPCs功能改变及衰老中的影响作用[J].中国海洋药物,2020,39(5):38-44.
岩藻黄素在振荡剪切应力致EPCs功能改变及衰老中的影响作用
The role of fucoxanthin on alteration of EPCs cell function and cellular senescence induced by oscillating shear stress
投稿时间:2020-07-23  修订日期:2020-09-09
DOI:
中文关键词:  岩藻黄素  振荡剪切应力  早衰  活性氧  内皮祖细胞
English Keywords:fucoxanthin  oscillating shear stress  cell senescence  reactive oxygen species  endothelial progenitor cells
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作者单位邮编
贺燕婷# 潍坊医学院 基础医学院 261053
李振凤# 潍坊医学院 基础医学院 261053
崔星慧 潍坊医学院 基础医学院 261053
于洁 潍坊医学院 基础医学院 261053
王美月 潍坊医学院 基础医学院 261053
成敏 潍坊医学院 基础医学院 261053
张晓芸 潍坊医学院 基础医学院 261053
崔晓栋* 潍坊医学院 基础医学院 261053
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中文摘要:
      目的 探讨岩藻黄素(Fucoxanthin, FUCO)对振荡剪切应力作用下内皮祖细胞(Endothelial progenitor cells,EPCs)生物学功能及细胞衰老的影响作用。方法 采用Flexcell STR-4000平行板流动系统模拟体内扰动流剪切应力(振荡剪切应力,Oscillatory shear stress,OSS,± 4 dynes/cm2,1 Hz)并加载于EPCs;利用结晶紫染色法观察OSS作用后EPCs的形态学改变;应用Boyden小室、自发性细胞贴壁及Matrigel基质胶分别检测细胞迁移、粘附和体外血管形成能力的变化;采用Western Blot检测促沉默信息调节因子1(Silent mating type information regulation 2 homolog-1,SIRT1)蛋白和β-半乳糖苷酶法检测细胞衰老程度的改变;通过Dihydroethidium(DHE)荧光探针法检测细胞内超氧化物阴离子。结果 与静态培养组相比,OSS导致EPCs形态学发生明显改变,表现为细胞长宽比降低,细胞之间间距增加;而FUCO对OSS导致的EPCs形态学改变无显著影响作用;与OSS处理组相比,FUCO可显著促进EPCs的迁移(10.00±1.73 vs 21.00±2.65个/视野)、粘附(8.00±1.00 vs 20.00±1.73个/视野)及体外血管形成能力(8538.67±1231.47 vs 11009.33±570.08像素/视野)。进一步研究发现,FUCO可降低OSS导致的β-半乳糖苷酶阳性细胞比例(21.18±1.38% vs 12.69±3.94%)以及促进SIRT1蛋白表达上调;同时,FUCO可减弱OSS诱导的EPCs胞内超氧阴离子的产生(P<0.05)。结论 FUCO可缓解OSS环境下引发的EPCs细胞功能降低、细胞衰老的发生,提示其可有效提升内源性EPCs的血管修复能力。
English Summary:
      Objective To investigate the role of fucoxanthin on the biological function and cell senescence of endothelial progenitor cells (EPCs) under oscillating shear stress (OSS). Methods The Flexcell STR-4000 parallel plate flow system was used to simulate the disturbed flow shear stress (represented by OSS, ±4 dynes/cm2, 1 Hz) and loaded on the EPCs; Alters of morphological on EPCs by oscillating shear stress were observed by crystal violet staining; Changes in migration, adhesive and angiogenesis properties of EPCs were be detected by Boyden chamber, spontaneous cell adherence and Matrigel matrix gel respectively. Cellular senescence detection is characterized by the expression of Silent mating type information regulation 2 homolog-1 (SIRT1) and β-Galactosidase activity. Dihydroethidium(DHE) as a fluorescent probe was used for the determination of intracellular superoxide radical anion. Results Compared with the static group, the morphology of EPCs were significantly changed in OSS group, which showed that the aspect ratio of EPCs was decreased while the spacing between cells was increased; Moreover, compared with the OSS treatment group, FUCO could significantly promote the migration (10.00±1.73 vs. 21.00±2.65 cells per field of vision), adhesion (8.00±1.00 vs. 20.00±1.73 cells per field of vision), and angiogenesis ability (8538.67±1231.47 vs 11009.33±570.08 pixel value per field of vision) of EPCs in vitro. Furthermore, we found that FUCO could reduce OSS-induced increase of β-Galactosidase positive cells (21.18±1.38% vs. 12.69±3.94%); Besides, FUCO also could promote the up-regulation of SIRT1 protein expression and attenuate intracellular superoxide anion production in EPCs induced by OSS (P<0.05). Conclusion FUCO could promote the cell function and down-regulation of cell senescence of EPCs in the OSS environment, which indicated that it would effectively improve the ability of endogenous EPCs to repair blood vessels.
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