| At present,the research of nanoscale drug delivery system is still a hot spot.Loading chemotherapy drugs into nanocarriers can reduce the side effects of drugs,increase their water solubility,prolong the circulation time in vivo,and increase the efficacy of drugs.In this paper,the Bovine Serum Albumin Nanoparticles(BSA NPs)loaded with Doxorubicin(DOX)and loaded with Gambogic Acid(GA)were prepared by desolvation-chemical cross-linking method.A series of physical and chemical characterization and optimization were carried out,and the anticancer activities of the DOX-BSA NPs(DNP)and GA-BSA NPs(GNP)were characterized in vitro and in vivo.The main work of this paper is as follows:Initially,DNP and GNP were prepared by desolvation-chemical crosslinking method.Using Minitab software,2~3 full factor experiments were designed.Taking the drug loading as the index,the preparation process of DNP and GNP was optimized by software analysis of the experimental results.In vitro drug release experiments showed that DNP and GNP had the characteristics of acid stimulated responsive release,which indicated that DNP and GNP could respond to release drugs in the more acidic environment of tumor cells.Secondly,the anticancer activities of DNP and GNP in vitro were investigated.The experimental results showed that the combination of DNP and GNP had synergistic antitumor effect.Firstly,the cytotoxicity of DOX and GA was investigated.DOX and GA could significantly inhibit the survival of HepG2 hepatoma cells,and the combination of the two drugs had synergistic effect according to different proportions.When the combination ratio of the two drugs was 1:1,it had the strongest synergistic effect,and the combination index(CI)was 0.22.Compared with the control group,the number of cells in G2/M phase increased from 7.73%to 16.01%and 10.22%in DOX group and GA group,respectively.The proportion of cells in G2/M phase increased from 7.73%to 21.36%in combination of two drugs.The results showed that the cell cycle could be blocked in G2/M phase by DOX and GA and their combination.The blank BSA NPs had almost no cytotoxicity.The combination of DNP and GNP had synergistic antitumor activity,and the synergistic effect was strong when the drug proportion was 1:1,CI was 0.38.Finally,the anticancer activities of DNP and GNP in vivo were investigated.It is further proved that the combination of DNP and GNP had synergistic effect in the treatment of tumor.The liver tumor model was established in nude mice.The fluorogram of the drug in each organ showed that the DOX was metabolized completely in each organ and tumor after 24 hours,but the content of DNP in the tumor site was increased compared with 4 hours.In vivo antitumor experiments showed that the combination of DNP and GNP had the greatest antitumor effect,and the combination of DNP and GNP was still better than the single drug when the dose was reduced to 1/3,indicating that the combination of DNP and GNP could enhance the antitumor ability.It was found that the systemic toxicity of DNP and GNP was lower in the experiment of body weight change of nude mice.HE staining showed that DNP,GNP and combination group had more tumor necrosis than DOX and GA group.DOX and DNP had obvious organ damage to heart,liver and spleen.c-Caspase-3 and Ki-67 staining showed that the comination of GNP and DNP could promote the apoptosis of tumor cells and inhibit the proliferation of tumor cells.In conclusion,DNP and GNP were successfully prepared by desolvation-chemical crosslinking method.DNP and GNP with high drug loading were prepared by using the optimized process with drug loading as the index.The combination of the two could synergistically inhibit the survival of tumor cells.In vivo experiments also showed that the combination of DNP and GNP could significantly inhibit the growth of tumor,with less toxicity.It provided the basis and foundation for further development and utilization of albumin nano drug carrier system for synergistic chemotherapy,and had a potential application prospect. |
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