Study On The Preparation Of COST@MSN-GA Nanoparticles And Their Activity In Treating Drug-induced Liver Injury

The incidence of liver disease is yearly growing and has become a major threatening to human health.In recent years,natural drugs have been widely used in the treatment of liver injury due to their advantages of multiple targets and low toxic side effects.Mesoporous silica nanoparticles（MSN）are widely used in drug delivery systems due to their strong stability,good biocompatibility,controllable particle size and pore size.Chitosan oligosaccharides with a molecular weight of 1000 Da（COST）has the potential of treating drug-induced liver injury due to its antioxidant and anti-inflammatory effect.In this study,MSN were used as an excellent carrier of COST in order to improve the mechanical strength,stability,targeted release ability and drug efficacy of COST.Dendritic MSN was synthesized by oil-water two-phase stratification method,and then modified with amino group（MSN-NH₂）.Finally,Glycyrrhetinic acid（GA）was modified on MSN to construct MSN-GA.The morphology characterization and thermodynamic properties were analyzed by a variety of instruments,such as transmission electron microscopy,scanning electron microscopy.The results showed that:the average particle size and Zeta potential of MSN,MSN-NH₂ and MSN-GA were 157.9 nm,164.4 nm,184.1 nm and-50.7 m V,-18.9 m V,-40.5 m V.The pore size and pore volume of MSN and MSN-GA were 12.54 nm,11.00 nm and 0.71 cm³/g,0.65 cm³/g.Compared with the infrared spectrum,the surface modification process of mesoporous silicon nanoparticles is feasible.MSN-GA nanoparticles loaded with COST（COST@MSN-GA）were prepared by solution diffusion adsorption method using MSN-GA as carrier and COST as model drug.Single factor investigation was carried out on the possible influencing factors of drug loading method.Expert-Design software was used to design the response surface method for the factors and optimize the optimal preparation scheme.The release of COST@MSN-GA was studied in vitro.The biocompatibility of COST@MSN-GA and COST was investigated by hemolysis test and cell scratch experiments.The results showed that the optimal drug loading process were 50℃reaction temperature,35h reaction time,M_COS:M_MSN-GA=1:1.Under such conditions,the EE%and DL%of drug loading nanoparticles were 44.1%and 78.2%.MSN-GA carrier has p H response performance,and has sustained release effect.COST@MSN-GA has good blood compatibility and cell compatibility,indicating that it has good biocompatibility.The activity of COST@MSN-GA in treating drug-induced liver injury was studied by cell experiment.First,LO2 cells were modeled for APAP-induced injury,and the low,medium and high doses of COST@MSN-GA were respectively determined to be 200μg/m L,400μg/m L and 800μg/m L.The intervention of COST,MSN and COST@MSN-GA on the model cells showed that COST@MSN-GA and COST increased the survival rate of LO2 cells.ALT and AST levels in the medium were measured.COST and COST@MSN-GA could reduce ALT and AST levels,and COST@MSN-GA had a better effect than COST.Flow cytometry was used to quantitatively analyze the uptake of MSN-GA and MSN by LO2 cells.The results showed that MSN-GA enhanced the uptake of drugs by LO2 cells compared with MSN,showing the receptor-mediated targeting effect.The mechanism of COS in the treatment of drug-induced liver injury was analyzed by network pharmacology method.By constructing the‘COS-targets-Drug-induced liver injury’network and‘Protein-Protein Interaction’network,we get the main core targets were ALB,AKT1,VEGFA,MMP9,CASP3,IGF1,SRC,EGFR,HRAS and NOS3.Through enrichment analysis of GO and KEGG pathways,it is concluded that COS is mainly involved in the regulation of cell adhesion,reaction to nutrient level,metabolism of reactive oxygen species and other biological processes,and may play a role in the treatment of drug-induced liver injury through lipids and atherosclerosis,PI3K/Akt signaling pathway,Rap1signaling pathway and other ways.Through organ localization analysis of the main core targets,the results showed that the core targets were mainly distributed in the heart,lung and liver,suggesting that chitosan oligosaccharides can be made into liver targeted drugs to play a better therapeutic effect of drug-induced liver injury.