| Since 2000,Chinese government carry out stringent policy of grain for green,which make confined farming took place of free-range grazing.The excessive use of high-energy feedstuff not only increases the cost of raising sheep,but also reduces the quality of mutton.In recent years,the ecological grass husbandry industry has gradually developed into a regional characteristic industry.As a Chinese traditional medicine,Apocynum venetum L.have an effects on lowing hypertention and blood lipid,and can be used as a functional forage to feed Tan-sheep by our pilot experiment.Based on the results of pre-experiment,using the latest bio-techniques,such as serum metaboiome and liver transcriptome,we researched the effects of A.venetum functional feedstuff on blood biochemical parameters,lipid metabolism and mutton quality from various levels,also,we explored its metabolic regulation mechanism.These studies showed that:1.A.venetum functional forage(shortened AV functional forage below)can increase the weight of Tan-sheep,improve blood biochemical indicators,better mutton quality,and increase the water content of mutton.A,B,C and D represented forage added AV leaves o%,5%,10%and 15%,respectively.The weight gain of the group B was the fastest,followed by the C group and the A group.The growth of the D group was the slowest.With the increasing of the AV content,the triglyceride content continued to decrease,and the longer the test period,the more significant the effect.At the end of the experiment,the C group reached the lowest level(1.62±0.07 mol/L);At the same time,the growth rate of total cholesterol slowed down,and the total protein and albumin fluctuated,and alkaline phosphatase and aspartate aminotransferase The activity of the AV functional forage can also increased the water retention capacity of the beach mutton and increased the cooked mutton rate.The cooked mutton rate of the C group was greater than of the A group,and the water loss rate was the lowest.2.AV functional forage caused significant changes in micro-molecule metabolites in the serum of Tan-sheep.The results showed that a total of 832 metabolites were detected in 60 samples(time series)in negative ion mode,and 8 differential metabolites were screened,among which p-chlorophenol was down-regulated,?-hydroxybutyric acid,pyrocatechol,p-chlorophenol,fumaric acid,ethylmalonic acid,dihydroxyphenylacetic acid,gentisic acid and protocatechuic acid are up-regulated;apocynum can induce carbon water such as tricarboxylic acid cycle,glycolysis,fructose,mannose metabolism and galactose metabolism Compound metabolic pathways,amino acid metabolism processes such as arginine,serine,threonine,tyrosine,and taurine metabolism,and changes in bile acid metabolism pathways.From the perspective of metabolomics,it is preliminarily expounded that A.venetum forage regulates the nutritional metabolism of Tan-sheep by promoting the carbohydrate metabolism and lipid metabolism,strengthening immunity and facilitating digestion.3.AV functional forage caused differential expression of lipid metabolism related genes in the liver of Tan-sheep.The results showed that 12 samples of transcriptome sequencing obtained 105.55 Gb Clean Data,and 1 766 new genes were discovered,of which 1018 were functionally annotated;the number and type of differentially expressed genes increased with the increase of A.venetum L.,and A Compared with the group,the number of DEG up-regulation in B,C,and D groups was 20,449,and 1.The number of down-regulated genes was 3,2 751,and 3 358,among which pyruvate dehydrogenase kinase 4(PDK 4)and acetyl-CoA acyltransferase 1(ACAA 1)and fatty acid binding protein 1(FABP 1)are up-regulated.malonyl-CoA decarboxylase(MLYCD)is down-regulated;in GO classification results,differentially expressed genes are mainly enriched in skeletal system development,organs Morphological development and tissue development,cell stress response,cytokine negative feedback regulation,triglyceride metabolism,phylogeny,cell regulation process and Golgi development,GO items;KEGG annotation results mainly involve PI3K-Akt signal transduction,AMPK Signal transduction,PPAR signal transduction and metabolic pathways such as fatty acid metabolism and degradation.From the level of gene transcription,the differentially expressed genes and metabolic pathways related to the metabolism of carbohydrates and fatty acids in the liver of Tan-sheep were analyzed.4.AV functional forage affected the lipid metabolism of Tan-sheep and involveed multiple metabolic pathways.The results showed that 821 metabolites were detected from 40 samples(additional difference)in the negative ion mode based on non-targeted metabolism technique,and 1766 new genes were extracted from 12 liver tissues by transcriptase sequencing,of which 1018 were obtained.Functional annotation;screening of 6 identifiable differential metabolites such as alanine,oxycholic acid,catechol,?-hydroxybutyric acid,gentisic acid and proline,respectively Associated with fatty acid binding protein 1,interleukin 2,interferon regulatory factor 5,ubiquitin-coupled enzyme E2 M,oxysterol-binding protein,and protein kinase C;involved in carbon metabolism,fatty acid metabolism,bile acid metabolism,tyrosine Metabolic,pantothenate and CoA biosynthesis and extensive metabolic pathways,combined analysis showed that A.venetum feedstuff mainly regulates the metabolism of Tan-sheep through enzymes and cytokines in the energy metabolism pathway.A.venetum feedstuff may regulate the nutritional metabolism of Tan-sheep by promoting the metabolic pathways such as the tricarboxylic acid cycle,bile acid metabolism and inhibition of fatty acid synthesis. |
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