Preparation And Biological Study Of Folic Acid-targeted Ginsenoside Rg5-loaded Bovine Serum Albumin Nanoparticles

Ginsenoside Rg5(Rg5),a triterpene saponins,extracted from natural herbal plant Ginseng,is one of the most potent anticancer drugs acting on various carcinoma cells.However,the therapeutic potential of Rg5 was limited by its poor bioavailability,and non-target delivery.Research workers are keen on the combination of nanomedicine and oncology under the context of nanotechnology.The abovementioned deficiencies of Rg5 could be overcome by targetable,biodegradable and biocompatible nanoscale drug delivery systems.So,we prepared folic acid(FA)functionalized Rg5 loaded Bovine serum albumin(BSA)nanoparticles(FA-Rg5-BSA NPs)to improve efficacy,and targetable delivery of Rg5.Based on this,the main experimental research results of this paper are as follows:(1)Rg5-BSA NPs were prepared by desolvation method,and folate molecules were modified with the active amino group on the surface of Rg5-BSA NPs by amidation reaction,to get FA-Rg5-BSA NPs.The preparation process of Rg5-BSA NPs and FA-Rg5-BSA NPs were optimized by single factor experiment.The final optimization results were:BSA concentration of 15 mg/mL,BSA solution pH of 9,water anhydrous ethanol volume ratio of 1:3,Rg5 and BSA molar ratio of 20:1,and ethanol addition rate of 1.5-2.0 mL/min,stirring speed 600 r/min,stirring time 24 h,the molar ratio of folic acid to BSA reaction was 15:1,after the addition of folic acid reaction time 6 h,centrifugal speed 10000 r/min.(2)The prepared FA-Rg5-BSA NPs had a hydrated diameter of 201.4 nm,a PDI of 0.081,and a zeta potential of negative.It has a small spherical shape and a smooth appearance.The drug loading was 12.64±4.02%,the encapsulation efficiency was 73.59±5.50%,and the folate coupling amount was 94.6 ?g/mg BSA NPs.The aqueous solution of FA-Rg5-BSA NPs can maintain stability for about 8 weeks at 4 ?.Under acidic(pH 5)conditions,Rg5 is released from the particles faster than in a neutral environment.(3)In vitro cell MTT toxicity assays demonstrated that FA-Rg5-BSA NPs have higher ability to inhibit cancer cell proliferation in MCF-7 cells compared to Rg5-BSA NPs and Rg5,whereas in normal L929 cells High cell survival rate.This indicates that FA-Rg5-BSA NPs have certain selectivity for different kinds of cells.By Hoechst 33342 and AO/EB fluorescence staining experiments,FA-Rg5-BSA NPs induced apoptosis in breast cancer MCF-7 cells compared with Rg5-BSA NPs and Rg5.Observing the uptake profile of FITC-labeled Rg5-BSA NPs and FA-Rg5-BSA NPs into MCF-7 cells,the amount of FITC-labeled FA-Rg5-BSA NPs was higher than that of FITC-labeled Rg5-BSA NPs.When free folic acid was added,the amount of FITC-labeled FA-Rg5-BSA NPs uptake by the cells was significantly reduced.This suggests that folate-modified nanoparticles can promote cellular uptake via the folate receptor-mediated endocytic pathway.(4)In the MCF-7 xenograft tumor model,in vivo pharmacodynamic studies showed that FA-Rg5-BSA NPs inhibited tumor growth more than Rg5-BSA NPs and Rg5,and had better antitumor activity in vivo.In vivo real-time bioimaging shows that FA-Rg5-BSA NPs have better tumor targeting than Rg5-BSA NPs.