| Ginsenoside compound K(CK)is a metabolite of protopanaxadiol ginsenosides,which is the main active compound in ginseng.It has many pharmacological properties and exhibits significant anticancer activity against various cancer cells,moreover,it can be used in the development of functional foods.However,ginsenoside CK has poor water solubility and low bioavailability,which limits its application to some extent.In order to solve this problem,we firstly prepared a chitosan nano delivery system based on amphiphilic deoxycholic acid-O-carboxymethyl chitosan and ginsenoside CK as a model drug.Subsequently,A54 was used as a targeting peptide to prepare a novel chitosan nano delivery system with liver cancer targeting.The structure and morphology of the two drug-loading micelles were characterized by particle size,zeta potential,dispersion coefficient,drug loading,encapsulation efficiency,and morphological observation.The in vitro release behavior of the two polymer micelles was studied by dialysis method,and the release mechanism of polymer micelles targeted with hepatocellar carcinoma was analyzed by curve fitting.The cytotoxicity of polymer micelles in vitro was determined by MTT assay.The apoptosis rate was detected by Annexin V-FITC apoptosis detection kit,and the cell uptake capacity of polymer micelles was evaluated by Cy5.5 as a fluorescent probe.The results are as follows:(1)The deoxycholate-O-carboxymethyl chitosan polymer micelles(CK-NPs)encapsulated with ginsenoside CK prepared by ultrasonic dialysis had a particle size of 172.1～192.3 nm,and the zeta potential of-24.7～-21.8 mV,the drug loading of 3.03～10.65%,and the encapsulation efficiency of 20.27～42.65%.Its shape was spherical structure,which met the requirements of nano delivery system and could be effectively delivered into tumor cells.Due to hydrogen bonding and electrostatic interaction,the CK-NPs showed good sustained-release behavior and pH-responsive release.In vitro experiments showed that ginsenoside CK and CK-NPs had a dose-dependent inhibition effect on HepG2 cells with IC50 values of 23.33 and 16.58 μg/mL,respectively.Apoptosis experiments showed that the apoptosis rates of HepG2 cells were 39.02%and 47.57%,respectively,after treatment with high concentrations of CK and CK-NPs,indicating that CK-NPs could promote apoptosis of tumor cells.Microscopic fluorescence imaging showed that CK-NPs could be taken up by HepG2 cells and had time-dependent uptake.(2)The A54 peptide modified PEGylated deoxycholate-O-carboxymethyl chitosan polymer micelle(APD-CK)targeted with hepatocellular carcinoma was successfully prepared by ultrasonic dialysis.The prepared APD-CK had a particle size of 156.8～171.4 nm,a zeta potential of-20.8～-18.4 mV,a drug loading of 1.61～3.18%,and an entrapment efficiency of 61.78～76.56%.The morphology was similar to spherical shape and the dispersion was good.The release of ginsenoside CK from APD-CK showed that the pH-responsive in vitro release behavior could be rapidly released at acidic pH.The drug release amount reached 73.49%within 144 h,and the sustained release effect was obvious.The release mechanism of the drug was non-Fickian diffusion.In vitro experiments showed that APD-CK significantly increased the uptake rate and apoptosis rate of human hepatoma HepG2 cells and Huh-7 cells,and the cytotoxicity of APD-CK was significantly higher than that of ginsenoside CK in the low concentration range(2.5-20 μg/mL).In this study,both novel drug-loaded micelles were showed higher cytotoxicity,apoptotic rate and cell uptake rate than ginsenoside CK.The results indicated that these two nano drug-loaded systems can be used as a potential nanocarrier to deliver hydrophobic anticancer drugs and improve the water solubility and permeability of drugs,which can enhance drug targeting and antitumor activity for clinical treatment. |
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