论文部分内容阅读
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(△f)与动态电阻变化(△R)响应。在所试H9C2浓度范围(5×10~4-4×10~5 cells/m L),△f与H9C2浓度呈线性关系,△R与H9C2浓度呈幂函数关系。我们用细胞粘弹性指数(CVI=△R/△f)来表征细胞的粘弹性。H9C2在异丙肾上腺素作用下,△f与△R增加、细胞-QCM表面黏附加强,细胞变硬;在维拉帕米作用下,△f与△R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。
Objective: In this study, real-time monitoring of the dynamic adhesion of rat cardiac myocytes (H9C2) to RGD self-assembled membrane containing cell adhesion recognition peptides by QCM was performed with two kinds of cardiovascular drugs Force, another negative muscle force) interaction. METHODS: Monolayers of 3-mercaptopropionic acid (MPA) monolayers were assembled on the gold electrode surface and the molecular adhesion molecules (KRGDs) were covalently coupled by amidation to form a dense self-assembled monolayer with specific adhesion to rat cardiomyocytes. QCM dynamically monitored the self-assembly of MPA / RGD and the cell adhesion of H9C2 cells on self-assembled membrane electrode in a dynamic and continuous manner. In addition, 20,000 H9C2 cells and the inotropic agent isoproterenol, the negative inotropic agent verapamil, were used to assess cell-to-cardiovascular drug interactions using QCM. RESULTS: Compared with the bare gold electrode, the gold electrode modified by MPA / RGD increased the QCM frequency shift (△ f) and the dynamic resistance change (△ R) response induced by H9C2 cell adhesion. The concentration of H9C2 in the concentration range of H9C2 (5 × 10 ~ 4-4 × 10 ~ 5 cells / m L) showed a linear relationship, △ R and H9C2 concentration showed a power function relationship. We used the cell viscoelasticity index (CVI = ΔR / Δf) to characterize the cell’s viscoelasticity. Under the action of isoproterenol, H9C2 increased △ f and △ R, the cell-QCM surface adhesion strengthened and the cells hardened; △ f and △ R decreased under the action of verapamil, the adhesion of QCM decreased, soft. CONCLUSION: QCM can be used to monitor the dynamic cell adhesion of rat cardiomyocytes in different concentrations, and can be used for the screening of cardiovascular drugs based on the ability of cell adhesion and cell viscoelasticity to distinguish positive and negative inotropic drugs.