Nanomanipulation And Its Application In Pharmacodynamics Of Chinese Medicines

Traditional Chinese medicine plays an important role in improving the immune function,inducing the apoptosis of tumor cells,inhibiting the tumor cell metastasis and enhancing the myocardial function.It is essential to find effective Chinese medicine efficacy evaluation methods for the development of Chinese medicine.However,there are some disadvantages when evaluating with optical microscopy,electron microscopy and other traditional biotechnologies due to the low resolution,complicated sample preparation and damage to the internal structure of cells.Nanomanipulation technology has the advantages of precision positioning,high resolution and real-time monitoring of cell dynamics in physiological environment.The interaction between drugs and cells is studied according to the cellular physics,which provides a new way for the drug efficacy evaluation.Cells are the most basic structural unit in the body.Biophysical properties can be used as novel markers because they are closely related to cell states.Furthermore,it can be used to characterize the cellular function and the response to the drug.Based on the nanomanipulation technology,multiple physical parameters such as cell morphology(dynamic changes of cell ultrastructure),elasticity and adhesion can be obtained under the stimulation of drugs,and then the behavior of drugs can be evaluated.The work is described in the following.1.Based on the AFM nanomanipulation technology,the interaction between the tip and the single cell is studied.The cantilever calibration and deformation detection methods are investigated.The mechanical contact models of AFM probe and cell interaction are established to perform the detection and characterization of the cells,which lays the foundation for the evaluation of drug efficacy.2.The evaluation method of mechanical properties for living cells is studied.A nanomanipulation system is developed to carry out the tasks for data acquisition,light spot position detection,coarse positioning,scanned imaging,contact point determination and acquisition of force curves.The changes in the elastic modulus of cells at different degrees of malignancy and at different loading rates are analyzed.3.The nanomanipulation technology is employed to investigate the effects of the American ginseng on cellular mechanical properties.The optimized hybrid mechanical model of probe-cancer cell is proposed to accurately determine the mechanical properties.The relationship between the dose and the time response of liver cancer cells to the drugs is quantitatively studied,and the changes in mechanical properties such as the cell morphology,roughness,height and cellular elastic modulus are obtained.The mechanical model of interaction between the probe and the single cardiomyocyte is established.The contractile force and beating frequency of cardiomyocytes after adding American ginseng water extract are acquired.The efficacy of the American ginseng is then evaluated with the nanomanipulation technology.4.The effects of cucurbitacin E on esophageal cancer cells with different differentiation stages are studied by nanomanipulation technology.The adhesion and elastic modulus of the esophageal cancer cells with different differentiation stages are obtained to evaluate the efficacy of cucurbitacin E.5.Nanomanipulation technology is used to implant TRAIL plasmids into the colon cancer cells.The process of the probe into the cells is analyzed.The green fluorescence is observed with a fluorescent inverted microscope.The results show that the TRAIL plasmids are implanted successfully using the nanomanipulation technology.Furthermore,the effect of TRAIL plasmid on colon cancer cells is evaluated.With the developed nanomanipulation system,the mechanical contact models of AFM probe and cell interaction are established.The quantitative changes in cell physical properties are detected after drug-cell interactions and the work lays the foundation for evaluating the efficiency of drugs.The results show that the nanomanipulation technology provides an important tool for drug efficiency evaluation,diagnosis,and drug and cell interaction at the molecular level,and it is helpful for the development of Chinese medicine and the treatment of diseases.