Intervention Effect Of Polysaccharides From Different Processed Products Of Angelica Sinensis On Chicken Liver Injury Induced By Ceftiofur Sodium Combined With LPS

Ceftiofur sodium is a kind of cephalosporin antibiotics used in veterinary clinic to prevent and treat chicken colibacillosis.When ceftiofur sodium kills E.coli,it releases endotoxin(also known as lipopolysaccharide,LPS).Whether the accumulation of endotoxin can cause complex liver injury is unclear.After liver injury,ceftiofur sodium may accumulate in livestock and poultry,resulting in drug residues,and then lead to food safety problems in animals.Firstly,the methods of establishing chicken liver injury model induced by ceftiofur sodium combined with LPS were screened,and the chicken liver injury model induced by ceftiofur sodium combined with LPS was established.The protective effects of different processed polysaccharides of Angelica sinensis on liver injury were studied by using different processed polysaccharides of Angelica sinensis.The details are as follows:1.Preparation and analysis of polysaccharides from different processed products of Angelica sinensis: Different processed products of Angelica sinensis were prepared according to the Processing Specification of Gansu Province Traditional Chinese Medicine.Crude polysaccharides from different processed products of Angelica sinensis were prepared by water extraction and ethanol precipitation,and refined polysaccharides were obtained by protein removal and dialysis.The yield of crude polysaccharides from different processed products of Angelica sinensis was wine-processed Angelica sinensis 6.46%>oil-processed Angelica sinensis 5.36%> unprocessed Angelica sinensis 5.25%>soil-processed Angelica sinensis 5.04%>charred Angelica sinensis 4.44%.After refining,the highest yield of polysaccharides was wine-processed Angelica sinensis 3.24%,the lowest yield of polysaccharides was unprocessed Angelica sinensis 1.92%.2.Screening of Chicken Liver Injury Model Induced by Ceftiofur Sodium and LPS: 350 1-day-old laying hens were purchased in 5 batches and fed freely and drank water.Screening of the dose and time of liver injury induced by Ceftiofur Sodium,screening of the dose of LPS-induced liver injury,single and combined use of Ceftiofur Sodium and LPS-induced liver injury,and combined use of Ceftiofur Sodium with different sources of LPS-induced liver injury were carried out respectively.The comparative study of injury and repetitive screening of time point and dose of liver injury induced by ceftiofur sodium combined with self-made LPS in chickens were conducted in 5 batches of animal experiments.After the experiment,the mental state of chickens in each group was observed,blood was collected and serum was prepared.The biochemical indexes related to liver injury,such as aspartate aminotransferase(AST)and glutamate pyruvate transaminase(ALT),were detected.The liver was dissected and fixed in 10% formalin for histopathological observation.The contents of AST and ALT in serum were detected.The results showed that the levels of ALT and AST in different doses of ceftiofur sodium groups increased significantly(P < 0.05),and there were different degrees of hepatocyte pathological changes,including vacuolar degeneration and inflammatory cell infiltration.When the doses of LPS were 2,4 and 6 mL/kg,the levels of ALT and AST increased significantly(P < 0.05).The hepatocyte injury in LPS(2 mL/kg)group was milder,and the hepatocytes in LPS(4 mL/kg)group had obvious vacuolar degeneration and steatosis.In LPS(6 mL/kg)group,hepatocyte necrosis was serious and there were almost no intact cells.Chickens died.Therefore,based on the commonly used dosage of ceftiofur sodium and the results of LPS screening,we determined that the dosage of ceftiofur sodium combined with LPS to replicate liver injury model was 5 mg/kg/d and 4 mL/kg,respectively.The results of screening the time point and dose of liver injury induced by ceftiofur sodium combined with LPS showed that compared with the normal control group,when the dose of LPS was 4 mL/kg,the liver injury was more obvious at 6 h and 48 h.3.Intervention effect of polysaccharides from different processed products of Angelica sinensis on chicken liver injury induced by ceftiofur sodium combined with LPS3.1 Study on dosage screening and intervention effect of raw Angelica polysaccharide: 1-day-old laying hens were randomly divided into normal control group,model group,low-dose and high-dose intervention group.Each intervention group was given different doses of unprocessed Angelica sinensis polysaccharide once a day for 10 consecutive days.According to the pre-screening method,blood was collected and liver was dissected 6 hours and 48 hours after the establishment of the model.The results showed that the high-dose intervention group of Angelica sinensis polysaccharide(0.2 g/kg/d)had better effect on preventing and treating liver injury.3.2 Effects of different processed polysaccharides of Angelica sinensis on liver injury induced by ceftiofur sodium combined with self-made LPS: 1-day-old laying hens were randomly divided into normal control group,model group,low and high dose intervention groups of processed polysaccharides(0.125,0.25 g/kg/d).Each intervention group was given different doses of polysaccharides from Angelica sinensis processed products once a day for 10 consecutive days.According to the pre-screening method,blood was collected 6 hours and 48 hours after the model establishment to prepare serum and liver was dissected.Total bilirubin,triglyceride,total cholesterol,high density lipoprotein and low density lipoprotein in serum were detected.ALT,AST,SOD and MDA were detected by the kit.The results showed that the polysaccharides from different processed products of Angelica sinensis could improve the liver injury induced by ceftiofur sodium combined with LPS in varying degrees.The main manifestations were reducing the contents of AST,ALT,γ-GT and MDA,increasing the levels of SOD,T-AOC and repairing the liver cell injury.In a word,polysaccharides from charred Angelica sinensis and soil-processed Angelica sinensis have better effect in preventing and treating liver injury.