The Improvement Effect Of ASH Water Extract On Liver Lipid Accumulation And Egg Production Performance In Elderly Laying Hens

Artemisia scoparia Herba(ASH),as a traditional Chinese medicine with the same origin as food and medicine,has the effect of protecting the liver and promoting the gallbladder.The excessive accumulation of liver fat in elderly laying hens can lead to a decrease in liver function,leading to a decrease in egg production performance.Network pharmacology plays an important role in analyzing the multi-components of traditional Chinese medicine.This study uses network pharmacology methods to study the treatment of liver fat accumulation in elderly laying hens with ASH water extract,improve egg production rate,and provide a basis for using Artemisia scoparia as a feed supplement.Method: 80 Hy-line Brown laying hens(70 weeks old)and 20 Hy-line Brown laying hens(30weeks old)were selected for the experiment.After one week of adaptive feeding,their weight,physical condition,and mental state were similar,and the experiment lasted for four weeks.There are 20 young chickens in the positive control group(35wk),and the elderly laying hens are randomly divided into four groups: control group(75wk),low-dose intervention group(75wk+L),medium dose intervention group(75wk+M),and high-dose intervention group(75wk+H).In the ASH extract intervention group,the doses were 1 g/chicken,2 g/chicken,and 4 g/chicken,respectively.During the experiment,record the egg production and measure the relevant egg quality.After the experiment,weigh the weight of each group of laying hens and collect serum samples from each group of laying hens.After dissection,observe the ovaries and follicles,separate the follicles and ovarian matrix,and measure the weight of ovarian matrix and grade of follicles.Observe the changes in liver color,morphology,and size,weigh the liver weight,and collect liver samples to make liver slices.Observe the pathological and histological changes of the liver with HE staining;Oil staining was used to observe the lipid accumulation in liver cells.Using serum samples,test liver function and blood lipids using a reagent kit.Protein immunoblotting was used to detect the expression levels of lipid metabolism related factors SREBP-1,FAS,and ACC in the liver.ELISA kit is used to determine the content of reproductive hormones in serum.By using network pharmacology methods to analyze the key targets and signaling pathways of ASH in the treatment of fatty liver,intersection genes of active ingredients and disease-related targets were obtained from relevant databases,and key genes were identified through PPI network analysis and pathway enrichment analysis.Protein immunoblotting and immunofluorescence were used to detect the expression of key genes AKT and P-AKT.Analyze the effect of ASH extract on excessive liver fat accumulation,and verify the molecular docking of components and central targets.The results are as follows:(1)ASH water extract can reduce the liver weight and body weight of elderly laying hens,and significantly improve liver function.The liver histology test showed that the ASH water extract significantly improved the liver lipid accumulation and hepatocyte steatosis of the aged laying hens.In addition,the levels of ALT and AST in the serum of 75 week old laying hens increased compared to 35 week old laying hens,while the levels of both in the serum of elderly laying hens in the groups treated with ASH water extract decreased(P<0.05),and liver function improved.(2)Compared with young laying hens aged 35 weeks,the liver and serum TC,TG,and LDL levels of laying hens aged 75 weeks significantly increased,while HDL levels decreased.After intervention with ASH water extract,the levels of TC,TG,and LDL were reduced,while the levels of HDL were increased(P<0.05).Meanwhile,the expression levels of lipid metabolism indicators,including FAS,ACC,and SREBP-1,decreased in the liver of elderly laying hens in the administration group,indicating that ASH water extract can improve liver fat accumulation in elderly laying hens.(3)Compared with 35 week old laying hens,the laying rate and egg quality of 75 week old laying hens decreased.However,after receiving ASH water extract,the laying rate of 75 week old laying hens was improved,and egg quality indicators such as Hastelli unit,eggshell thickness,protein height,and yolk weight were all improved.The ASH water extract had a significant effect on eggshell strength,protein height,and Hastellin units(P<0.01),with a trend to increase egg yolk weight and eggshell thickness(P<0.05).The follicle diameters of F1,F2,and F3 in each group treated with ASH water extract were significantly longer than those at 75 weeks(P<0.05).During this process,the levels of reproductive hormones,including FSH,LH,E2,and VTG,in the serum of elderly laying hens were also increased and improved(P<0.05).(4)In terms of network pharmacology,13 active ingredients were extracted from the database and analysis platform of Systems pharmacology of traditional Chinese medicine.And conduct GO and KEGG enrichment analysis,use PPI network interaction to screen key therapeutic targets AKT,and verify molecular docking of components and central targets.Meanwhile,the experimental results showed that ASH extract reduced liver fat accumulation by inhibiting phosphorylated AKT cleavage.Conclusion: The intervention of ASH water extract in laying hens during the late laying period can improve liver fat accumulation,increase egg production rate,and egg quality.The key target of network pharmacology analysis,AKT,inhibits the synthesis of liver fat.ASH water extract,as a medicinal and edible homologous traditional Chinese medicine,can be used as a feed supplement for laying hens to improve the decrease in egg production rate caused by liver lipid accumulation in the late laying period.