高脂饮食对大鼠主动脉内皮细胞形态及刚度的影响
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1.北京航空航天大学生物与医学工程学院 生物力学与力生物学教育部重点实验室;2.北京航空航天大学生物医学工程高精尖创新中心

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国家自然科学基金项目(面上项目,重点项目,重大项目)


Effect of high fat diet on Rat aortic endothelial cell morphology and stiffness
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1.Key Laboratory for Biomechanicsand Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University;2.Advanced Innovation Center for Biomedical Engineering;3.Beihang University;4.Advanced Innovation Center for Biomedical Engineering, Beihang University

Fund Project:

The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)

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    摘要:

    摘要:目的:研究高脂饮食对大鼠主动脉内皮细胞形态与刚度的影响。方法:利用植块法获得大鼠胸主动脉内皮细胞,在倒置显微镜下观察细胞形态,通过荧光染色比较两组细胞F-actin平均荧光强度,利用原子力显微镜(AFM)检测细胞刚度。结果:细胞在植块后第7天有少量细胞爬出,第14天细胞融合成单层,VIII因子染色呈阳性,证实为内皮细胞。高脂饮食引起细胞周长(AS: 145.4±65um vs. CON: 111.9±44um, P<0.01)、长轴/短轴比率(Aspect ration,AR)(AS: 3.2±1.8 vs. CON: 2.4±1.3,P<0.01)、面积(AS: 554±259um2 vs CON: 542±224um2,P>0.05)显著增大;细胞圆度(AS:0.4±0.18 vs. CON: 0.6±0.19,P<0.01)则相对减小。细胞F-actin平均荧光强度为:AS:60±2.9 vs. CON: 54±1.6,P<0.01。AFM显示高脂饲养引发内皮细胞刚度显著增大(AS: 5.2±2.2Kpa vs. CON:4.3±2.2Kpa,p<0.01)。结论:高脂饮食将引发内皮细胞形态细长化改变,刚度增大,这可能与高血脂导致的血管功能异常直接相关,使其成为动脉粥样硬化的重要诱因。

    Abstract:

    Abstract: Objective : In the present study, we are attempting to investigate the high fat diet effect on endothelial cell morphology and stiffness. Methods: Rat aorta endothelial cells (RAECs) were obtained from rat thoracic aorta by explants adherent method and identified by factor VIII immunofluorescence staining. Cell morphology was observed under inverted microscopy and quantified by shape parameters including circularity, aspect ratio, perimeter, and area. The mean fluorescent intensity of F-actin was calculated to see the alteration of cell cytoskeleton. Cell stiffness was measured by atomic force microscopy (AFM). Results: The endothelial cells migrated from tissue plant and form confluence after 14days cultivation. The morphology results indicated that cells from high fat diet rats show enhanced perimeter (AS: 145.4±65um vs. CON: 111.9±44um, P<0.01), aspect ratio (AS: 3.2±1.8 vs. CON: 2.4±1.3,P<0.01), and area (AS: 554±259um2 vs CON: 542±224um2,P>0.05), while less circularity (AS: 0.4±0.18 vs. CON: 0.6±0.19,P<0.01) compared with the control cells. This may be resulted from more F-actin filaments (AS:60±2.9 vs. CON: 50±1.6,P<0.01) expressed in AS cells. The cell stiffness of the high fat feeding group was significantly higher than that of the normal feeding group (5.2±2.1Kpa VS 4.3±2.2Kpa, p<0.01). Conclusions: High fat diet will change the morphology and stiffness of endothelial cells, which may subsequently affect their normal function, being more vulnerable to atherosclerotic risk factors.

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  • 收稿日期:2019-09-20
  • 最后修改日期:2019-11-18
  • 录用日期:2019-11-19
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