Preparation,characterization And Activity Evaluation Of Resveratrol-loaded Glycyrrhizin Conjugated To Human Serum Albumin Nanoparticles With Liver Targeting

Resveratrol(RES)is a non-flavonoid polyphenols containing stilbene structure,which has certain anticancer activity to hepatocellular carcinoma cells.However,resveratrol has poor water solubility,poor stability,low bioavailability,and short half-life in vivo characteristics limit its exhibit excellent biological activity in vivo.Studies have shown that glycyrrhizic acid has a biological target on the surface of the liver,because the surface of the liver cells have glycyrrhizin receptor,so the glycyrrhizic acid and human serum albumin coupling into nanoparticle carrier(GL-HSANPs),can increase the drug's active target delivery.(GL-HSA-RESNPs)conjugated with glycyrrhizic acid was prepared by the method of high-pressure homogenization emulsification.The purpose of this study was to prepare the recombinant medium for the expression of resveratrol.The characterization of the RES nano-targeting formulation improves the RES bioavailability.In this study,GL-HSA-RESNPs nanoparticles were prepared by nanotechnology.In the preparation process optimization,the particle size was an important criterion for selecting the optimal conditions.The effects of the volume of human serum albumin,the volume ratio of aqueous phase to organic phase,the ratio of homogenization speed and homogenization time,homogenization pressure and homogeneity on particle size were investigated.The laser particle size analyzer was used to detect GL-HSA-RESNPs particle size;Scanning electron microscopy and transmission electron microscopy(TEM)were used to investigate the surface morphology of nanoparticles;The encapsulation efficiency,drug loading and in vitro release properties of resveratrol nanometer preparation were detected by high performance liquid chromatography;The solid characterization of nanoparticles was investigated by Fourier transform infrared spectroscopy,X-ray diffraction,differential scanning calorimetry and thermogravimetric analysis;The inhibitory effect of GL-HSA-RESNPs on HepG2 cells and the hepatic targeting of in vivo nanoparticles were investigated by MTT and in vivo targeting experiments;Study on the bioavailability and in vivo tissue distribution of resveratrol nanoparticles in rat tail vein.The results are as follows:1?The glycyrrhizin-conjugated human serum albumin nanoparticles were detected by infrared spectroscopy and the characteristic peaks,and the absorption peak of amide bond increased,indicating that glycyrrhizic acid and human serum albumin coupled successfully.The glycyrrhizin-conjugated human serum albumin nanoparticles and the conjugated amount of glycyrrhizic acid were 112.56 ?g/mg.2?Orthogonal test and single factor experiment were used to select the optimal conditions:human serum albumin concentration was 4 mg/mL,the volume ratio of water phase to organic phase was 9:1,the homogenization speed was 8000 rpm,homogenization time was 5min,homogeneous pressure to 800bar,homogeneous number of 9 times.The average particle size of GL-HSA-RESNPs prepared under the optimal conditions was 108.1 ± 5.3 nm,and the encapsulation efficiency and drug loading were 83.6%and 11.5%,respectively.After SEM observation,nanoparticles showed uniform dispersion of spherical.Resveratrol was expressed in glycosic acid-conjugated human serum albumin nanoparticles in an amorphous form by solid state analysis.In vitro studies on the release of GL-HSA-RESNPs showed a sustained release of the drug,and the water solubility was significantly improved.3?The effect of resveratrol nanoparticles on the inhibition and targeting of hepatocellular carcinoma cells in vitro and the validation of liver targeting experiments in vivo showed that GL-HSA-RESNPs and RES drugs were treated with HepG2 cells for 48 hours of the lethal doses of 62.5 ?g/mL and 95.5 ?g/mL,respectively,so GL-HSA-RESNPs will have a better therapeutic effect.The fluorescence intensity of the samples was GL-HSA-RESNPs>HSA-RESNPs>GL and HSA-RESNPs>RES,and the GL-HSA-RESNPs have certain targeted active;and distributed to the trend detected in vivo drug murine model of orthotopically transplanted H22 by small animal in vivo imaging device.Cy5-reactive dye-labeled GL-HSA-RESNPs was injected into the tail vein of the liver cancer model mice,and the fluorescence intensity of the liver was large and persistent compared with the healthy mice injected with the same dose,and the GL-HSA-RESNPs were significantly of liver cancer targeting.4?Pharmacokinetics of resveratrol nano-drug in rat tail vein injection:The concentration of resveratrol in the rats in the experimental group was higher than that in the control group.The half-life of the resveratrol nanoparticles in the experimental group was 2.997 h after the tail vein injection,and the biological half-life t1/2? was 8.778 h.The distribution half-life tl/2a of resveratrol powder in the control group was 1.794h and the biological half-life tl/2? was 5.501 h.The average residence time(MRT)of the nano-drug group in rats was 1.01 times of that of the original drug group.Resveratrol nanoparticles in the body to eliminate the rate is higher than resveratrol powder.The AUC value of resveratrol nanoparticles in rats was 1.7 times that of resveratrol.From the experimental results,the bioavailability of resveratrol nanoparticles was significantly improved.5?The concentration of resveratrol was measured at the time of 0.5,1,3,6,12 and 48 h at different time.The results showed that the absorption of nano-drugs in the main organs was significantly higher than the original drug,especially in the liver of the drug accumulation is obvious,compared with the original drug in the liver drug absorption is significantly different,nano drugs in the liver The concentration of accumulation and accumulation of far more than the original drug,indicating that resveratrol nanoparticles have significant liver targeting.The MRT of resveratrol nanoparticles in the liver was 1.5 times that of resveratrol.Resveratrol nano-powder in the liver AUC value of resveratrol powder is 4.17 times.Resveratrol nano-powder has a targeting to rat liver tissue,which means that resveratrol nanotechnology is expected to become a new type of injection for liver disease.