Protective Effect Of Angelica Polysaccharide On Copper Induced Liver Injury And Behavioral Changes

As one of the essential nutrients in animals,copper plays an important role in physiology.In modern animal production and breeding,copper,as an important metal element in animal diet,is used to promote animal growth and development,improve animal production performance,improve feed absorption and conversion rate,and achieve good economic benefits.At the same time,excessive use of copper leads to copper poisoning in livestock and poultry from time to time.Copper poisoning not only affects the physiological and mental state of animals,but also affects the health of animals,And it pollutes the environment of animal husbandry.The daily use of copper in different animal diets should be strictly controlled,the behavior of livestock and poultry should be observed,combined with liver function and serological indicators to determine whether it is copper poisoning,and timely use of drugs for treatment.Angelica polysaccharide,as a widely used traditional Chinese medicine,has the effects of improving anemia,anti-tumor,improving immune function,protecting liver and anti-oxidation.Copper is mainly distributed in the liver.Excessive copper will cause toxicity to liver and other organs and cause oxidative stress reaction.It can not only protect the liver and protect the liver,but also relieve the oxidative stress caused by excessive copper,reduce the anxiety of animals,improve the mental state,and reduce the loss of modern animal husbandry due to copper poisoning.Therefore,in this study,the rat and rabbit models of copper induced liver injury were established by feeding rats and rabbits with copper loaded diet,and then the rats and rabbits were given different doses of Angelica polysaccharide solution by gavage,and then two behavioral experiments,Barnes maze and elevated plus maze,were carried out to detect the spatial discrimination cognitive ability,learning memory and anxiety degree of copper poisoning rats,The blood biochemical indexes,liver peroxidase activity and liver tissue structure of rats and rabbits were detected to explore the reasonable intake of copper in the diet of rabbits,the changes of behavior,blood biochemical indexes and liver peroxidase activity when copper was excessive,the protective effect of Angelica polysaccharide on copper induced liver injury in rats and rabbits,and the intervention effect of behavior,Objective to provide a scientific basis for the use of Angelica polysaccharide to alleviate copper induced liver injury and behavioral cognitive changes.The test is divided into three parts:Experiment 1The effect of Angelica polysaccharide on the behavior of copper induced rats In order to study the effect of Angelica polysaccharide on copper induced behavioral changes in rats,40 1-month-old SD rats were randomly divided into normal control group,model control group,Angelica polysaccharide 1 group(ASP1),Angelica polysaccharide 2 group(ASP2)and Angelica polysaccharide 3 group(ASP3).The rats in control group and ASP1,2 and 3 groups were fed with powder feed containing 1 g / kg copper sulfate and deionized water containing 0.185% copper sulfate respectively.The normal control group was fed with the same dose of common feed and deionized water for12 weeks.From the 7th week,ASP1,2 and 3 groups were fed with copper sulfate powder feed and deionized water each time,The rats in the normal control group and the model control group were given the same volume of normal saline by gavage with different doses of ASP solution(100mg / kg / d,200 mg / kg / d,400 mg / kg / d).After the completion of copper loading diet and Angelica polysaccharide gavage,the rats in each group were subjected to elevated cross maze and Barnes maze test.Before Barnes test,the rats were given 6-day adaptive training,and the behavioral indexes were recorded;After 12 weeks of copper loading diet and Angelica polysaccharide gavage,the elevated plus maze test and Barnes maze test were continued to record behavioral indicators.The results showed that the percentage of open arm residence time and the percentage of open arm entry times in the model control group were significantly lower than those in the normal control group.The number of errors,latency and distance of target hole in ASP3 group were significantly higher than those in normal control group,and the treatment effect of ASP3 group was the best.The results showed that Angelica polysaccharide could improve the behavioral changes caused by copper poisoning,improve the spatial cognitive ability and memory of copper poisoning rats,and reduce the anxiety of copper poisoning rats.Experiment 2 The effect of Angelica polysaccharide on blood biochemistry and liver tissue of rats induced by copper In order to study the effect of Angelica polysaccharide on blood biochemistry and liver tissue of copper induced rats,the blood of abdominal aorta was taken and the serum was separated.The activities of ALT and AST were measured by automatic biochemical analyzer,and the values of MDA,SOD and GSH-Px were measured by spectrophotometer.Small pieces of liver tissue were cut and washed in PBS buffer(p H 7.3).Paraffin embedded sections were prepared by hematoxylin eosin staining;The activities of antioxidant enzymes(MDA,SOD,GSH-Px)in liver tissue were detected by spectrophotometer;The contents of serum copper and liver copper were detected by atomic absorption spectrophotometer.The results showed that the copper content in serum and liver tissue of the model control group was significantly increased,indicating the success of the model.The activities of ALT and AST in the model control group were significantly increased,while the activities of ALT and AST in the Angelica polysaccharide treatment group were gradually decreased,which were more and more close to the normal control group.The SOD and GSH-Px values in the model group were significantly lower than those in the normal control group,while the MDA content was significantly higher.The SOD and GSH-Px values in the treatment groups ASP1,2 and 3were significantly higher than those in the model control group,while the MDA content was significantly lower.In the normal control group,the structure of liver tissue was complete and the structure of hepatic lobule was clear.In the model control group,the liver tissue was damaged,the structure of hepatic lobule was fuzzy and inflammatory cell infiltration appeared.The structural integrity of liver cells in ASP1,2 and 3 groups was improved compared with that in the model control group.The results showed that copper poisoning inhibited the activities of antioxidant enzymes and led to lipid peroxidation.Angelica polysaccharide could inhibit the lipid peroxidation induced by copper poisoning,improve the activities of antioxidant enzymes in serum and liver,and protect the liver injury caused by copper poisoning.Experiment 3 The effect of Angelica polysaccharide on blood biochemistry and liver function of New Zealand rabbits induced by copper In order to study the effect of Angelica polysaccharide on blood biochemistry and liver function of rabbits induced by copper,20 New Zealand rabbits aged 6 months were randomly divided into normal control group,model control group,Angelica polysaccharide 1 group(ASP1),Angelica polysaccharide 2 group(ASP2),and Angelica polysaccharide group(ASP3).After one week of adaptive feeding,the New Zealand rabbit model control group and ASP1,2,and 3groups were fed with sulfuric acid respectively.After the 4th week,ASP1,2,and 3 groups were given ASP solution of different concentrations(50mg/kg,100mg/kg,200mg/kg)to rabbits after feeding copper sulfate powder feed and deionized water,350mg/kg copper sulfate feed and 0.185% copper sulfate deionized water,and the normal control group for12 weeks.From week 7,ASP1,2,and 3 groups were fed with copper sulfate powder feed and deionized water each time.Normal control group and model control group were given equal volume of normal saline.After abdominal aorta blood was taken,serum was separated,and the activities of ALT and AST were measured by multifunctional automatic biochemical analyzer,MDA,SOD,and GSH-Px were measured by spectrophotometer.Thepartiallivertissuewascutandwashedwith PBS buffer(p H7.3),whilegrindingwith 9timesofhomogenatemedium.Afterwards,themixturewascentrifugedat 3500r/minfor 10 min,preparing 10%livertissuehomogenateandputtingitinrefrigeratorat ﹣ 80 ℃ fordetectionof MDA,SOD,and GSH-Pxinthetissue.The contents of serum copper and liver copper were detected by atomic absorption spectrophotometer.The results showed that the copper content in serum and liver of model control group was significantly higher than that of normal control group,and there was no statistical difference between Angelica polysaccharide treatment group and model control group.The activity of ALT and AST in plasma was significantly higher than that of model control group,and ALT and AST activity in ASP1,2,and 3 groups were significantly lower than that in model control group,and AST value was close to normal value.The SOD and GSH-Px values in the model group were significantly lower than those in the normal control group,while the MDA content was significantly higher.The SOD and GSH-Px values in the treatment groups ASP1,2,and 3 were significantly higher than those in the model control group,while the MDA content was significantly lower.The biochemical indexes of liver and serum in ASP3 group were closer to normal values,indicating that the dosage of 200 ml/kg/d Angelica polysaccharide solution given to the New Zealand rabbits poisoned by copper could protect the liver injury caused by lipid peroxidation.