| Alcohol is a common drink in our daily work,but unreasonable drinking will cause ace-r tain degree of damage to the body and lead to many negative effects.In recent years,diseases s-u ch as hepatitis,liver cirrhosis and acute liver injury are on the rise all over the country,which pose a serious threat to people’s lives and health.Curcumin(Cur)has hepatoprotective effect.In thispaper,Cur was used as an experimental drug to explore the protective effect of Cur/m PEG-PBL A micelle on alcoholic liver injury in vitro.The specific research contents and results are as foll-o ws:1.The preparation methods of Cur/m PEG-PBLA micelles were screened and the prescription was optimized.It was determined to be prepared by dialysis method,the organic solvent was dimethyl sulfoxide(DMSO),the volume ratio of water to solvent(DMSO)was 2.5:1,and the feed ratio was 20:1.2.The morphology of micelles was observed by transmission electron microscope(TEM),and its stability and hemolysis were investigated.And verify whether the preparation of Cur into micelles can improve the problem of low solubility and poor stability of the drug.The results showed that the average particle size of Cur/m PEG-PBLA micelles was 120～130 nm,Zeta potential was(-8.17 ± 1.26)m V,PDI was 0.20±0.014,and the shape was spherical.Cur/m PEG-PBLA micellar sol-ution can improve the stability of curcumin and reduce the degradation rate of the drug.The micellar solution has good dilution,storage and plasma stability.Through the observation of c olor andthe calculation of hemolysis rate,it is confirmed that the polymer micelle does not have h emolysis,does not destroy the red blood cells in the blood,and has good blood compatibility.3.A method for the determination of Cur content was established.The drug loading(DL)and entrapment efficiency(EE),of Cur/m PEG-PBLA micelles were calculated and their release invitro was tested.The standard curve is Y=208727X-78088(R~2 = 0.9999),and there is a good linear relationship in the linear range.The precision,stability,method recovery and sample recovery are all in line with the requirements.The drug loading is(4.87 ± 0.42)% and the entrapment efficiency is(73.26 ± 3.16)%.In the in vitro release experiment,the Cur/m PEG-PBLA micelle showed a slow cumulative release of the drug,with a cumulative release of only 28.94% in 10 h and 48.25% in 48 h,which had a good sustained release effect.4.Establishment of cell injury model.Two kinds of hepatoma cells(Hep G-2)and normal hepatocytes(7702)were selected and anhydrous ethanol was used as inducer.The cell survival rat e was measured and calculated by MTT method.The Hep G-2 cell injury model was initially selected with 400mmol/L anhydrous ethanol for 12 h,and the cell survival rate was 70.82%.7702 the cell survival rate was 73.13% when the cell model was made at 400 mmol/L for 12 h.In order t ofurther verify the reliability of the modeling conditions,the values of ALT and AST in the cell s upernatant were determined by kit under these conditions.The results showed that the ALT and A ST values of the Hep G-2 cell model group were 1.7 times and 1.95 times of that of the controlgr oup,and the ALT and alt values of the 7702 cell model group were 2.27 times and 2.75 timeshig her than those of the control group.It was verified that the model of hepatocyte injury was su-ces sful,and 400 mmol/L anhydrous ethanol for 12 hours was selected as the best condition for mode ling.5.The protective effect of Cur/m PEG-PBLA micelles on alcoholic liver injury was studiedfrom two aspects of prevention and repair.The results showed that curcumin solution and Cur/m PEG-PBLA micelle solution had no toxic effect on the two kinds of cells when the concentration of curcumin solution and cur/m PEG-PBLA micelle solution were 1～10 μg/ml.6.Hep G-2 cells: in terms of preventive effect,when the micelle concentration was 2～10 μg/ml,there was significant difference between the model group and the model group.When the c oncentration was 10 μg/ml,the cell survival rate reached 108.04%,which was more significant than that of the model group,and the prevention effect of cell damage was the most obvious;when the concentration of curcumin solution was 1～10 μg/ml,there was obvious difference between the model group and the curcumin solution concentration of 10 μg/ml The cell survival rate was 109.87%,which was more significant than that of the model group(P < 0.001);When the concentration of curcumin solution was between 2～10 μg/ml,the cell survival rate was 93.78%,which was moresignificant than that of the model group;when the concentration of curcumin solution was 2～10 μg/ml,the cell survival rate was increased in a dosedependent manner;when the concentration of Cur solution was in the range of 4～10 μg/ml,compared with the model group The cell survivalrate was 97.36% when the concentration was 10 μg/ml,which was more significant than that oft he model group(P < 0.001).In conclusion,the two solutions can prevent and repair Hep G-2 cell injurymodel,and the higher the concentration,the more obvious the effect.7.7702 cells: in terms of preventive effect,when the concentration was 2～10 μg/ml,the c ell survival rate was significantly improved compared with the model group,and there were significant differences between the two groups in a dose-dependent manner.When the micelle concentration was 10 μg/ml,the cell survival rate reached 101.95%,which was more significant compared with the model group;when the concentration of curcumin solution was 2～10 μg/ml,the cell survival rate was improved,and there was a significant difference compared with the model group.I n the repair effect,when the micelle concentration was 4～10 μg/ml,the cell survival rate was significantly improved compared with the model group,and the difference was significant and in a d osedependent manner.When the micelle concentration was 10 μg/ml,the cell survival rate reached90.23%,which was significantly different from the model group.When the concentration of curcu min solution was 4～10 μg/ml,the cell survival rate was significantly different from that of the model group(P < 0.001).In conclusion,the two solutions can prevent and repair 7702 cell injury m-odel,and the higher the concentration,the more obvious the effect. |
