薯蓣皂苷元丝素蛋白纳米粒的制备、表征及其对前列腺癌荷瘤裸鼠肿瘤的抑制作用OA

Objective To prepare diosgenin silk fibroin nanoparticles(Doi-SF-NPs)and evaluate their physicochemical properties,tumor targeting and in vivo anti-tumor effects.Methods The main influencing factors of Doi-SF-NPs were investigated by single factor experiments,and Box-Behnken design-response surface methodology(BBD-RSM)was employed to optimize prescription process of Doi-SF-NPs.Doi-SF-NPs was characterized by transmission electron microscopy(TEM)and Fourier transform infrared spectroscopy(FT-IR).Drug release behavior of Doi-SF-NPs in phosphate buffer saline solution with different pH values was investigated.MTT method and Annexin V/PI double staining method were used to investigate the inhibitory and proapoptotic effects of Doi-SF-NPs on PC3 cells,respectively.The nude mouse of prostate cancer was established,compared the concentrations of diosgenin and Doi-SF-NPs in tumor tissues,and then evaluated the tumor targeting ability of Doi-SF-NPs using the relative uptake rate and ratio of peak concentration.The nude mouse model of prostate cancer was divided into model(normal saline)group,positive control(cisplatin,2.0 mg/kg)group,diosgenin(10 mg/kg)group,low-dose Doi-SF-NPs(5 mg/kg)group and high-dose Doi-SF-NPs(10 mg/kg)group,the in vivo anti-tumor effects of Doi-SF-NPs was compared with diosgenin.Safety evaluation of Doi-SF-NPs was also performed.Results Optimal prescription process of Doi-SF-NPs nanoparticles as follows:The mass ratio of silk fibroin to diosgenin was 11.94∶1,the volume ratio of the organic phase to aqueous phase was 5.99∶1,and the preparation time was 2.75 h.Average envelopment efficiency,drug loading,particle size and ζ potential were(84.07±0.70)％,(6.41±0.05)％,(164.29±4.43)nm and(-23.01±1.17)mV,respectively.Diosgenin formed Doi-SF-NPs with silk fibroin through hydrogen bonding.The appearance of Doi-SF-NPs was spherical nanoparticles.Doi-SF-NPs existed higher cumulative release in phosphate buffer solution of pH 5.5 and 6.0,which demonstrated that Doi-SF-NPs had obvious pH-sensitivity.MTT method results showed that IC50 of diosgenin and Doi-SF-NPs on PC3 cells were 72.44 and 55.68 μmol/L,respectively.Doi-SF-NPs more effectively induced apoptosis of PC3 cells than that of diosgenin.The relative uptake rate of Doi-SF-NPs in in tumor tissue was 3.23,and the peak concentration ratio was 2.60,which demonstrating that Doi-SF-NPs could significantly increase the accumulation in tumor tissue.Doi-SF-NPs(10 mg/kg)increased the tumor inhibition rate of diosgenin(10 mg/kg)from 29.47％to 66.18％.Doi-SF-NPs did not affect the major organs of tumor-bearing nude mouse,which showed that the safety of Doi-SF-NPs was high.Conclusion Doi-SF-NPs could improve the tumor targeting of diosgenin and significantly enhanced anti-tumor efficacy.Therefore,Doi-SF-NPs had the potential to be developed into an antitumor drug.