| The contamination of food and feed by aflatoxin B1(AFB1)is widespread in the world and has become a global problem endangering food safety and livestock and poultry farming.At present,there is a lack of safe and effective drugs for the prevention and treatment of AFB1 poisoning.Curcumin is a plant polyphenol extracted from turmeric,which has various pharmacological effects such as anti-inflammatory,antioxidant,and anti-tumor.Its medicinal value and therapeutic significance have become a research hotspot in recent years.However,the solubility of curcumin is poor,which greatly reduces the oral bioavailability,which seriously restricts the application scope of curcumin.Liposome is an emerging drug embedding technology in recent years.It has the advantages of high drug loading efficiency,high stability in biological environment,high bioavailability and controllable quality,which makes it better than other carriers in clinical research.application prospects.In this study,curcumin liposomes were first prepared by thin film dispersion method combined with homogenization technology,and the encapsulation efficiency was used as an indicator to optimize the formulation process by single-factor investigation and response surface methodology.properties,structural properties,appearance,stability,etc.;then,based on the establishment of high performance liquid chromatography(HPLC)to detect the content of curcumin in rabbit plasma,taking ordinary curcumin as the reference preparation,and using the curcumin prepared in this experiment Curcumin liposomes were tested preparations,and the blood pharmacokinetic characteristics and relative bioavailability of the two preparations were studied in rabbits;it provided a theoretical basis for evaluating the clinical effectiveness and rational drug use of curcumin liposomes;In this study,an in vitro AFB1-induced AML-12 hepatocyte injury model was established,and the toxicity of AFB1 on AML-12 cells and the intervention effect of curcumin liposomes were investigated by group comparison method.The results of the study are as follows:(1)Preparation and evaluation of curcumin liposomes:Taking the encapsulation rate as the evaluation index,the curcumin liposomes were successfully prepared by thin film dispersion method combined with homogenization technology.After optimization and verification,the best prescription is:cholesterol content of 25 mg,curcumin content of 12 mg,and buffer volume of 25 m L;the encapsulation efficiency of the prepared liposomes is 96.6%;the appearance is spherical or ellipsoid,which is a uniform A stable dispersion system with a particle size of 128 ± 1.5 nm and a potential of +1.10 ± 0.09 m V;the structure and properties of curcumin liposomes were explored by infrared spectroscopy,and it was found that the liposomes were formed by hydrophobic interaction of phospholipid bilayers.Curcumin was embedded;the in vitro release results showed that compared with free curcumin,curcumin liposomes had a sustained release effect(free curcumin 24 h release was 75.92%,curcumin liposome 24 h release was 75.92%)42.02%).The stability results of curcumin liposomes showed that liposome entrapment significantly improved the stability of curcumin at 4 ℃ in the dark.(2)Bioavailability of curcumin liposomes in rabbits:An HPLC method was established for the determination of curcumin content in rabbit plasma.Plasma samples were extracted with ethyl acetate,detected by HPLC UV detection method,and quantified by external standard method.The methodological results showed that the standard working solution of curcumin in rabbit plasma had a good linear relationship in the range of 0.05-10 μg/m L(correlation coefficient>0.99),and the detection limit and quantification limit were 0.05 μg/m L and 0.1 μg/m L,respectively.,the standard addition method of curcumin with three concentrations(0.05,0.5,10 μg/m L)was used to investigate the precision and accuracy of the method.The intra-assay and inter-assay coefficients of variation were 6 % and 11 %,respectively.The average recovery rate in the range of 84.4 % to 117.3 % with good stability.12 mature male rabbits were randomly divided into 2 groups,and the reference preparation and the test preparation were orally administered to the two groups of rabbits at a dose of 100 mg/kg body weight,respectively.Blood was collected from ear vein at 1,1.5,2,3,4,and 6h.After pretreatment,the plasma samples were determined by the HPLC method established above,and analyzed by Winnonlin 5.2.1 software.The results showed that the Cmax of curcumin liposomes was significantly higher than that of the curcumin group,indicating that the absorption effect of the liposomes was significantly improved after encapsulating the drug;at the same time,the Tmax(2h)of the curcumin liposomes group was significantly later than that of the curcumin group.The AUC of curcumin liposome group(1906.31±111.87 ng·h/m L)was significantly higher than that of curcumin group(288.33±32.19 ng·h/m L),and the relative bioavailability in rabbits was 680.02 %.(3)In vitro pharmacodynamics study of curcumin liposomes:An in vitro AFB1-induced AML-12 hepatocyte injury model was established,and the cells were divided into AFB1 group,curcumin liposome control group,curcumin liposome intervention group,The curcumin intervention group,the curcumin control group and the blank control group were analyzed and compared with the supernatant transaminase(ALT,AST)activity,ROS content,inflammatory factors(TNF-α,INOS,IL-6,IL-1β)expression,to study the toxicity of AFB1 to AML-12 cells and the effect of curcumin liposome intervention.Compared with the blank control group,the serum transaminases of the AFB1 group were significantly increased(P < 0.01),the ROS content was significantly increased(P < 0.01),and the inflammatory factors(TNF-α,INOS,IL-6,IL-1β)The expression level of AML-12 was significantly up-regulated(p value),indicating that AML-12 hepatocytes were damaged and inflamed under the action of AFB1.Compared with the AFB1 control group,the serum transaminase levels in the curcumin intervention group and the curcumin liposome intervention group were significantly decreased by 25.94 % and 31.54 %,respectively(P<0.01),and the ROS content was significantly decreased(P<0.01).The expression levels of factors(TNF-α,INOS,IL-6,IL-1β)were all decreased,indicating that curcumin liposomes can significantly alleviate or antagonize AFB1-induced cell damage and inflammation,and its intervention effect is comparable to that of ordinary curcumin.Slightly better.In summary,the preparation method and prescription of curcumin liposomes were determined in this study,and curcumin liposomes with uniform particle size,stable system and high encapsulation efficiency were obtained;the relative biological properties of the liposomes in rabbits were obtained.The utilization degree was 680.02 %,and it had a good protective effect on AFB1-induced hepatocyte injury.The research results provided a solid theoretical basis for the development and clinical application of curcumin liposomes. |
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