Preparation Of Resveratrol Liver Targeted Liposomes And Its Preventive Effect On Hepatic Fibrosis In Rats

Objective:resveratrol is a natural anti-tumor,anti-cardiovascular disease,anti-mutation,antibacterial,anti-inflammatory,antioxidant,induction of apoptosis and estrogen regulatory substance,it is one of the main active components of Chinese traditional medicine knotweed.Hepatic fibrosis is a slow abnormal proliferation of connective tissue in the liver caused by a variety of pathogenic factors,which is a significant feature of more serious liver disease.If liver fibrosis exists for a long time and continues to develop,it may eventually become severe liver cirrhosis and liver cancer.According to theory,liver fibrosis can be reversed,but if the pathogenic factors always exist,it will eventually develop into irreversible liver cirrhosis.Resveratrol can protect the liver by inhibiting acute liver injury and anti-fibrosis to treat these reversible early liver diseases and prevent them from developing into irreversible liver cirrhosis.The prevention and treatment of liver fibrosis before liver cirrhosis should be studied.Recent studies have found that resveratrol plays an anti-fibrosis effect by reducing oxidative stress.Therefore,early anti-fibrosis therapy can effectively reverse this process and become a research hotspot.Methods:(1)Through single factor experiment,the encapsulation rate was used as the measurement index,and three key factors were selected to optimize the prescription and ultrasonic processing technical parameters.The optimal prescription was selected to prepare liver targeted liposomes,common liposomes,and free solutions.The liver target liposomes of resveratrol were characterized.(2)In the targeting experiment in vitro,the qualitative study of cell uptake was carried out by flow cytometry,and the cell activity was detected by CCK-8 method.(3)in the in vivo targeting experiment,a small animal imager was used to verify the distribution of resveratrol in rats at different time points.(4)The preventive effect of resveratrol liver targeted liposome was verified by the rat model of hepatic fibrosis induced by compound factors.That is,except for the normal control group,the other four groups were subcutaneously injected with 40%CCL4soybean oil solution,the first dose was 5ml/kg,and the rest were injected with 3ml/kg twice a week.At the same time,the rat model of liver fibrosis was made by feeding simple corn flour(80%corn flour+20%lard)in the first two weeks,mixed with 0.5% cholesterol and 30% ethanol as the only drink for four weeks.Resveratrol liver targeted liposomes,resveratrol ordinary liposomes and free resveratrol solution were injected into the tail vein in the process of modeling.Liver function index(ALT,AST,ALP,TP,ALB,GLOB,A/G),SOD,MDA,HYP,HA,LN,PC?,C-IV and pathological sections of three groups of different administration solutions were selected as the examination indexes.The preventive effects of resveratrol liver targeted liposome,resveratrol liposome and free resveratrol solution on liver fibrosis in rats were compared.Results:the optimal prescription was determined as follows:drug-lipid ratio 1:25,phosphorus-bile ratio 10:1,ultrasound power 150 W for 10 min.The average particle size of targeted liposomes was 120 nm,and the average entrapment efficiency was more than 83%.Resveratrol liposome has no hemolysis,meets the requirements of Pharmacopoeia and can be used for intravenous injection.In the in vitro targeting study,the fluorescence intensity of resveratrol liver targeted liposomes was the highest,indicating that the target modified liposomes did increase cell uptake compared with the other two groups.In the experiment of detecting cell activity by CCK-8 method,resveratrol liver-targeted liposomes can effectively bind to cells due to the modification of galactoside liposomes,and the liposomes release the drug slowly,giving full play to the long-term and effective inhibitory effect of resveratrol on HSC-T6 cells.In the in vivo targeting verification experiment,the distribution of drugs in animals was clearly seen by small animal imager,in which the longest duration of fluorescence signal was observed in rats injected with resveratrol liver targeted liposomes.it was observed that the fluorescence intensity of the liver of rats injected with resveratrol liver targeted liposomes was the highest compared with the other two groups,indicating that the content of resveratrol in liver tissue was the highest.In the study of the preventive effect of resveratrol liver targeted liposome on hepatic fibrosis model in rats,the results showed that the experimental rat hepatic fibrosis model was successfully established.In the liver function index,compared with the normal control group,the TP,GLOB,AST,ALT in the model control group was significantly higher and the ALB,A/G was significantly lower than that in the normal control group.Compared with the model control group,TP,GLOB,AST,ALT,in resveratrol liver targeted liposome group,resveratrol common liposome group and free resveratrol group decreased significantly and ALB,A/G increased significantly.Compared with resveratrol liver targeted liposome group,AST,ALT,in resveratrol liposome group was significantly higher than that in resveratrol liver targeted liposome group.There was no significant difference in GLOB,TP,but ALB,A/G decreased significantly.GLOB,AST,ALT,in free resveratrol group was significantly higher than that in control group(p<0.05.There was no significant difference in),TP,but ALB,A/G was significantly decreased in free resveratrol group.AST,ALT in free resveratrol group was significantly higher than that in resveratrol liposome(p< 0.05);ALB,).There was no significant difference in TP,A/G,GLOB.On the whole,the liver function indexes of each group were within the range of normal reference value.Compared with the normal control group,the biochemical indexes of liver tissue in the model group showed a significant decrease in SOD and an increase in MDA,HYP in the model control group,and SOD in resveratrol liver targeted liposome group,resveratrol common liposome group and free resveratrol group were significantly higher than those in the model control group(p<0.05).MDA,HYP was significantly decreased in resveratrol liver targeting liposome group and free resveratrol liposome group.Compared with resveratrol liver targeted liposome group,SOD in resveratrol liposome group decreased significantly(p<0.05),HYP increased significantly,but MDA had no significant difference,SOD decreased and MDA,HYP increased significantly in free resveratrol group.Compared with the resveratrol liposome group,the MDA,HYP in the free resveratrol group was significantly higher than that in the resveratrol liposome group,but there was no significant difference in SOD.Compared with the normal control group,HA,LN,PC? and CI-?in the model group increased significantly,while HA,LN,PC? and CI-Vdecreased significantly in the resveratrol liver targeted liposome group,resveratrol common liposome group and free resveratrol group.Compared with resveratrol liver targeted liposome group,HA,LN in resveratrol liposome group was significantly higher than that in resveratrol liver targeted liposome group,while HA,PC? and LN,CI-? in free resveratrol liposome group were significantly higher than those in free resveratrol liposome group.Compared with the resveratrol liposome group,there was no significant difference in HA,LN,PC? and CI-? in the free resveratrol group.In liver tissue,histological sections showed that resveratrol could reduce the deposition of collagen fibers and protect the liver,especially the liver targeting liposome of resveratrol.Conclusion:the liver targeting liposomes of resveratrol with liver targeting function were successfully prepared by this preparation process with good encapsulation rate and drug loading and no hemolytic activity.It has a preventive effect on experimental liver fibrosis in rats.