Studies On Synergistic Effect Of High Voltage Pulsed Electric Field Combined With EPA On A-549 Cells

Cancer is one of the major diseases endangering human health,especially lung cancer,its incidence rate is extremely high,and the mortality rate ranks first.In recent years,pulsed electric field electroporation technology has achieved remarkable results in tumor treatment.Compared with traditional tumor treatment methods,it has certain advantages and is currently a research hotspot in tumor treatment.Most of the previous studies on electroporation tumor treatment were tumor ablation by irreversible electroporation and electrochemotherapy by reversible electroporation.However,the tumor ablation area of irreversible electroporation is limited,and the traditional chemotherapeutic drugs used in electrochemical therapy have great toxic and side effects on the human body.How to reduce the damage to the human body as much as possible in tumor treatment is of great significance.In recent years,studies have shown that the anticancer components of traditional Chinese medicines can exert specific toxic effects on cancer cells,and are very weak to normal human cells.However,the cell membrane has a selective permeability mechanism,which hinders the exogenous drug molecules,which leads to the low intracellular drug concentration and it is difficult to reach the effective concentration when the anticancer components of traditional Chinese medicine are used for cancer treatment.Based on the above characteristics,this paper chose to use reversible electroporation generated by high-voltage microsecond pulsed electric field combined with traditional Chinese medicine ingredient eicosapentaenoic acid to act on lung cancer A-549 cells to study the synergistic effect of the two.Firstly,the simulation of single-cell dielectric model is carried out.The finite element analysis software COMSOL Multiphysics was used to establish a three-layer dielectric model of a single cell.The simulation revealed the law between the development of the cell membrane transmembrane potential and the electroporation of the cell membrane under the action of the microsecond pulsed electric field with different electric field strengths,as well as the intrinsic relationship between the pore density distribution and the changes in the conductivity and permeability of the cell membrane.Provide a theoretical basis for subsequent experimental research.Secondly,the effect of pulsed electric field combined with eicosapentaenoic acid on A-549 cells was studied.The effects of pulsed electric fields of different field strengths on cells and the effects of different concentrations of eicosapentaenoic acid on cells were respectively studied.Appropriate parameters were screened to study the effect of pulsed electric field combined with eicosapentaenoic acid on A-549 cells.The perforation rate of A-549 cells under the action of pulsed electric field was detected by PI staining method,and the activity and apoptosis rate of A-549 cells were detected by CCK-8 and Annexin V/PI double staining method.The morphological changes of cells under the action of pulsed electric field were observed by transmission electron microscope.It was verified that the microsecond pulsed electric field can significantly enhance the cytotoxic effect of eicosapentaenoic acid on lung cancer A-549 cells.Finally,the protein expression of A-549 cells under the action of pulsed electric field combined with eicosapentaenoic acid was studied.The protein factors with high correlation with lung cancer A-549 cells were selected,and the up-regulation and down-regulation of related protein expressions were detected by Western blot.It is proved that the microsecond pulse electric field combined with eicosapentaenoic acid can regulate the protein expression of lung cancer A-549 cells,and can inhibit the activity of cancer cells and induce the apoptosis of cancer cells at the protein level.In conclusion,this paper studies the mechanism of electroporation induced by microsecond pulsed electric field through simulation analysis.Then,through experimental research,it was verified that the microsecond pulsed electric field significantly enhanced the toxic effect of eicosapentaenoic acid on lung cancer A-549 cells from the cellular level and the protein level,which provided an important promotion method for the subsequent treatment of cancer with traditional Chinese medicine ingredients.