Study On The Real-time Monitoring And Drug Protection Of Vascular Endothelial Cell Injury

Cells are the basic structure and functional units of organism,cellular research is one of the basic links to conquer disease and reveal the mysteries of life.Aiming at the cells cultured in vitro,this paper of cellular behaviors and morphological changes of cell adhesion,growth,proliferation and injury in different interfaces was carried out,using piezoelectric sensing technology and electrochemical methods.The main contents are as follows:1.Cysteine and methionine were immobilized on QCM gold electrode,respectively,to construct two different electrode surfaces:the hydrophilic and hydrophobic surface.The adhesion,growth and H2O2-induced oxidation injury of human umbilical vein endothelial cells(HUVEC-C)on the two electrode surfaces were investigated by using the amino acid-modified electrodes based on cyclic voltammetry(CV),electrochemical impedance spectroscopy(EIS)technique,microscope observation and MTT measurement.The results indicate that the hydrophilic surface was helpful to promote the cell adhesion and proliferation but the hydrophobic surface did just the opposite.The strong oxidizability of H2O2 could result in the serious oxidation injury of the adherent cells.The cell-damage extent on the hydrophilic surface was greater than that on the hydrophobic surface,which was closely related to the worse adhesion and growth performance of HUVEC-C on the methionine-modified electrode.2.The modified glassy carbon electrode with poly(L-lysine)and graphene quantum dots(GCE/PLL/GODs)was prepared by using the electropolymerzation of L-lysine and the drop casting of GQDs.Electrochemical behavior of quercetin on this electrode was investigated based on cyclic voltammetry(CV)and the experimental results show that the current peaks could reach the maximum when 100-cycle CV scan,200-s quiet time and pH 6.0 B-R buffer solution were used.The electrochemical detection of quercetin with different concentrations was performed via differential pulse voltammetry(DPV).It is found that the DPV anodic peak current was proportional to the quercetin concentration from 0.05 to 5.0 ?M.The proposed electrochemical sensor was successfully applied in the determination of quercetin in Ginkgo dipyridamole injection and the satisfactory recovery values were obtained.3.The adhesion,spreading,and proliferation of human umbilical vein endothelial cell line(HUVEC-C)cells,on a gold electrode were monitored using quartz crystal microbalance(QCM)measurements.The viscodensity effect caused by the normal action of the cells led to a decrease of the resonant frequency and increase of the motional resistance.The oxidative injury of HUVEC-C cells appeared immediately with the addition of H2O2,exhibiting the decline of cellular spreading area and cell coverage on the electrode surface and resulting in inverted QCM responses.The injured extent of the cells was found to be related to the content of H2O2.It is found that 0.05 mM quercetin added beforehand in the growth medium could remove completely the oxidative action of 1.0 mM H2O2.Quercetin with increased dosage still exerted a partial protective effect on HUVEC-C cells against oxidative injury induced by 2.5 mM H2O2.The microscope observations,electrochemical measurements,and MTTanalysis validate the QCM assay results,indicating that quercetin is a valuable flavonoid anti-oxidant in the precaution and treatment for the oxidative injury of vascular endothelium.