| 周珍 周铁安 单梅 黄复深 周书岳 苏招红.QCM 监测自组装膜上心肌细胞黏附及其与心血管药物作用[J].,2016,16(1):58-62 |
| QCM 监测自组装膜上心肌细胞黏附及其与心血管药物作用 |
| Real Time Monitoring of Cardiomyocytes Adhesion and Cells-CardiovascularDrug Interactions Using RGD-Modified QCM |
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| DOI： |
| 中文关键词: 心肌细胞 细胞黏附 石英晶体微天平 细胞黏弹性 心血管药物 |
| 英文关键词: Cardiomyocytes Cell adhesion Quartz crystal microbalance Cell viscoelasticity Cardiovascular drugs |
| 基金项目:国家自然科学基金面上项目（21275048）；湖南省科技厅重点项目(2013TT1009) |
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| 中文摘要: |
| 目的：本试验采用石英晶体微天平（QCM）实时监测大鼠心肌细胞（H9C2）在含有细胞黏附识别多肽RGD自组装膜上的动
态黏附过程及随后与两种心血管药物(一种正性肌力、另一种负性肌力)相互作用。方法：在金电极表面自组装3-巯基丙酸（MPA）
单层膜，并经酰胺化共价耦合细胞黏附分子KRGD，形成对大鼠心肌细胞有特异性黏附的致密分子自组装膜。QCM以动态持续
的方式实时监测MPA/RGD 自组装及其不同浓度梯度H9C2细胞在自组装膜金电极上的细胞黏附过程。此外，选用20,000 个
H9C2 细胞和正性肌力药物异丙肾上腺素、负性肌力药物维拉帕米，用QCM评估了细胞-心血管药物的相互作用。结果：与裸金
电极相比，MPA/RGD修饰金电极增大了H9C2细胞黏附所引起的QCM频移(Df)与动态电阻变化(DR)响应。在所试H9C2 浓度
范围(5× 104 - 4× 105 cells/mL)，Df 与H9C2 浓度呈线性关系，DR 与H9C2 浓度呈幂函数关系。我们用细胞粘弹性指数(CVI=
DR/Df)来表征细胞的粘弹性。H9C2 在异丙肾上腺素作用下，Df与DR增加、细胞-QCM表面黏附加强，细胞变硬；在维拉帕米作
用下，Df与DR 降低、细胞QCM表面黏附减弱，细胞变软。结论：QCM可用于不同浓度大鼠心肌细胞的动态细胞黏附监测，并可
基于其细胞黏附与细胞黏弹性测定能力区分正性与负性肌力药物而可望用于心血管药物的筛选。 |
| 英文摘要: |
| Objective:Quartz crystal microbalance (QCM) was used for the dynamic monitoring of cell adhesion in real time of rat
myocardial cells (H9C2) onto self-assembled film containing cell recognition peptide RGD, as well as the subsequent interactions with
two cardiovascular drugs (one positively inotropic and the other negatively inotropic).Methods:A self-assembling 3-mercaptopropionic
acid (MPA) monolayer was formed on the gold QCMelectrode surface, allowing for subsequent covalent coupling with the cell adhesion
peptide KRGD, to form a dense molecular self-assembled monolayer with specific recognition motif for myocardial cells. QCMwas successfully
used for the dynamic monitoring of the MPA/RGD self-assembly process and the subsequent myocardial cell adhesion process
in the concentration range of 5× 104 - 4× 105 cells/mL of H9C2 cells in real time. Additionally, QCMwas used to assess the cells-cardiovascular
drug interactions using 20,000 H9C2 cells and positive inotropic drug, isoprenaline and negative inotropic drug, verapamil.Results:Compared to bare gold electrode, the MPA/RGD-modified gold electrode yielded enhanced frequency shift (Df) and motional resistance
change (DR) with the adhesion of H9C2 cells. In the tested H9C2 concentration range (5× 104 - 4× 105 cells/mL), Df was linearly related
to the cell concentration, whereas DR was a power function of the cell concentration. Cell Viscoelastic Index (CVI= DR/Df) was used to
characterize the cells' viscoelasticity. Under the effect of positive inotrope, isoprenaline, cell-QCMsurface adhesion became stronger with
increased Df and DR; CVI increased and the cells became stiffer. Whereas under the effect of negative inotrope, verapamil, cell-QCM
surface adhesion became weaker with decreased Df and DR; CVI dropped and the cells became softer.Conclusion:QCMcan be used to
monitor dynamic cell adhesion of different concentrations of H9C2 rat myocardial cells; and differentiate the effects of positive and negative
inotropic drugs based on changes in cell adhesion strength and cells' viscoelasticity, demonstrating its potential as a screening tool for
cardiovascular drugs. |
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