Nanoparticle Delivery Of Artesunate Enhances The Anti-tumor Efficiency By Activating Mitochondria-Mediated Cell Apoptosis

ObjectiveLiver cancer is a common malignant tumor,which has the characteristics of strong invasion,rapid growth of tumor,high rate of anti-chemotherapy and postoperative recurrence,which affects human health worldwide.Therefore,it is urgent and necessary to develop a safe and effective method to combat tumor.Recent studies have shown that artemisinin and its derivatives are considered to have a wide range of anti-tumor activities,and that artemisinin and its derivatives have obvious anticancer effects on tumor cells.Because its low concentration of artesunate mainly depends on tumor-induced tumor cell death,its antitumor effect is unfavorable and limited.In order to obtain better anti-tumor effect,artesunate from plant was used in this experiment.Ester,using bovine serum albumin as the carrier,preparation of artesunate bovine serum albumin nanoparticles.Artesunate bovine serum albumin nanoparticles promote the rapid release of cytochrome C by up regulation of related apoptotic proteins,destroy the integrity of mitochondria and activate mitochondria mediated apoptosis.SMMC-7721 and PLC cell lines are used as the research object,Conclusion artesunate bovine serum albumin nanoparticles can significantly promote cell apoptosis.MethodsThe optimal prescription was 500 ?g/mL of artesunate solution,35 mg/mL of bovine serum albumin solution and 30 drops /min of artesunate solution.The encapsulation rate and drug loading rate of the nanoparticles obtained by the optimal prescription were 82.4% and 13.3%,respectively.After characterization and analysis of Ats-BSA-NPs,the Ats-BSA-NPs under TEM presented a complete spherical shape with a dispersion index of 0.016 and an average particle size of 99.9 2.3nm.The results showed that the nanoparticles were uniformly dispersed in a good state.The Zeta potential of the nanoparticles was negative,with a magnitude of-25.6 4.3 mV.Compared with the rapid release of free drugs in vitro,the release rate of Ats-BSA-NPs is relatively slow,and due to the continuous degradation of Ats-BSA-NPs,it shows a stable and lasting release mode.MTT assay showed that compared with free artesunate,Ats-BSA-NPs had stronger cytotoxicity and higher sensitivity to smmc-7721 tumor cells and PLC tumor cells,and had stronger inhibitory effect on tumor cells.The distribution and localization of nanoparticles in the two tumor cells were observed by confocal microscopy,and the results showed that Ats-BSA-NPs could target the mitochondria and seriously damage the mitochondrial membrane,leading to depolarization of the mitochondrial membrane.After tumor cells were treated with free drugs and Ats-BSA-NPs for a certain period of time,the intracellular ROS content also increased,showing a time-dependent relationship.In addition,the cells treated with Ats-BSA-NPs produced significantly more ROS,indicating that Ats-BSA-NPs had stronger lethality.To investigate the correlation between cell death and Ats-BSA-NPs induced apoptosis,the expression of apoptotic proteins was detected by western-blot.The results showed that artesunate bovine serum albumin nanoparticles accumulated in mitochondria,Activated mitochondrial mediated apoptosis effect,and increased the expression of caspase-3 and caspase-9 related apoptosis proteins,leading to obvious apoptosis.ResultsArtesunate bovine serum albumin nanoparticles obtained by the method of dissolvation were spherical,uniform in size,and slow in release under electron microscope.The results of MTT and flow cytometry showed that artesunate bovine serum albumin nanoparticles had stronger cytotoxicity and higher sensitivity than those of free artesunate.According to the results of laser confocal scanning microscopy,the results of BSA NPS mediated transport of Ats to mitochondria.Western-Blot and Ros showed that,compared with the free artesunate bovine serum albumin(BSA),it was possible to transport its products to mitochondria.Western Blot and Ros.Rice grains have more obvious effect on promoting apoptosis.ConclusionsArtesunate bovine serum albumin nanoparticles prepared by the method of desolvation have the advantages of uniform particle size,good stability,good slow-release effect and no sudden release effect.Compared with free artesunate,artesunate bovine serum albumin nanoparticles have higher cytotoxicity.Artesunate loaded bovine serum albumin nanoparticles promote the rapid release of cytochrome C by up-regulating apoptosis-related proteins.Damage the integrity of mitochondria and activate apoptosis mediated by mitochondria.