Regulation And Mechanism Of Anthocyanins On The Browning Of White Fat In Animals At High Altitude Environment

With the improvement of living standards,people's demand for high-quality meat is becoming more and more urgent,and high-quality meat mainly depends on two important factors: muscle development and fat deposition.In order to better control the body fat deposition and improve the heat production of livestock adipose tissue,through technical means intervention or livestock genetic improvement can make the body rich white fat into brown fat to generate heat is one of the feasible ways.Promoting the browning of white fat is helpful to control the lipid deposition ability of the body,and provides a theoretical basis for improving the lean meat rate of livestock and poultry;At the same time,white fat browning can improve the cold resistance of animals,especially the cold resistance of newborn piglets,reduce production costs and improve economic benefits by improving their survival rate.In livestock production,it is expected to promote the white fat browning to prevent metabolic diseases in livestock and poultry,and provide a theoretical basis for the prevention and treatment of human obesity and type 2 diabetes.There are few reports on the fat deposition law of animals in high altitude environment and natural drugs such as anthocyanins in high altitude environment,the transformation of white fat into brown fat and its thermogenic mechanism.In this study,C57BL/6J mice and the primary adipocytes of huzhu bamei pigs were used to study the effects and mechanisms of different altitudes and anthocyanin concentrations on white fat browning by means of molecular biology and network pharmacology;Screening the action targets of anthocyanins in order to provide new ideas for improving lean meat rate,improving meat quality of livestock and poultry,and preventing human obesity and related metabolic diseases.The main research results are as follows:1.Regulation of different altitudes on Browning of animal white fatWith the increase of altitude,the body weight(P<0.01)and food intake(P<0.05)of mice decreased significantly,the contents of TC(P<0.05),TG(P<0.001)and LDL(P<0.01)in serum decreased significantly,the content of HDL(P<0.05)increased significantly,and the contents of i NOS(P<0.05),COX-2(P<0.05),IL-6(P<0.05)and HIF-1?(P<0.05)content decreased significantly;The level of prohibitin-1 was significantly increased(P<0.05),and the expression of perilipin-1 was significantly decreased(P<0.05);The level of white fat browning related genes UCP1 and PGC-1? and fndc5 and PRDM16 increased significantly(P<0.05).The effect of high-altitude environment was further verified in huzhu bamei porcine primary adipocytes.The results were consistent with the experimental results of mice at different altitudes.The above results show that high altitude environment can increase the number of mitochondria,improve lipid metabolism,activate the browning process of white fat,improve heat production,and reduce the incidence of obesity.2.Anthocyanins can promote the Browning of white fat in animals at high altitudeAnthocyanins significantly decreased the body weight(P<0.05),the contents of TC,TG and LDL in serum(P<0.05),and significantly increased the level of HDL(P<0.05),inflammatory factors i NOS,COX-2,IL-6 and HIF-1? in a dose-dependent manner at high altitude.The results showed that anthocyanins could improve the lipid metabolism of obese mice at high altitude;Further experiments showed that with the increase of anthocyanin concentration in the experiment at high altitude,the number of mitochondria in i WAT increased,lipid droplets shrank,fat cells shrank,and the level of mitochondria Prohibitin protein and m RNA related to mitochondrial formation increased significantly(P<0.05),The protein levels of white fat brown-related genes UCP1,PGC-1?,fndc5 and NRG4 were significantly increased in i WAT and BAT(P<0.05).In simulated high-altitude environment,pig primary adipocytes were treated with different concentrations of anthocyanins.The results were consistent with the experiment of obese mice.It further showed that anthocyanins could improve the process of animal lipid metabolism,promote the production of mitochondria,inhibit the aggregation of lipid droplets,accelerate the combustion and utilization of brown fat,and promote the browning process of animal white fat.3.Screening the key genes of anthocyanin promoting the Browning of white fat in animals at high altitude.Through high-throughput sequencing technology,transcriptome sequencing was performed on i WAT of obese mice and normal mice treated with high concentration of anthocyanin in high-altitude environment,and the differentially expressed genes were screened.GO,KEGG and protein interaction network analysis were performed on the differentially expressed genes.The results showed that anthocyanins regulate the occurrence of obesity,improve energy metabolism,and induce the Browning of white fat mainly through inflammatory pathways in adipose tissue.4.Screening the action targets of anthocyanins based on network pharmacology.Based on Pharm Mapper,Gene Cards and Dis Ge NET databases,27 anthocyanins such as EGFR,ADAM17 and LEP were identified,and then String database and Cytoscape3.7.2software were used to analyze the above targets.GO and KEGG pathways were analyzed to construct a target-signaling pathway-biological process network diagram,and the mechanism of anthocyanin regulating lipid metabolism was explored by using network pharmacology.Results The common targets of anthocyanin-obesity involved 60 signaling pathways,including PI3K/Akt signaling pathway,c GMP/PKG signaling pathway,FOXO signaling pathway,insulin resistance and AMPK signaling pathway.In this study,anthocyanins mainly regulate gene expression in obese mice through ADAM17 mediated PI3K/Akt signaling pathway,and activate the process of adipose tissue Browning.5.Mechanism of anthocyanins regulating the Browning of white fat in animals at high altitude.By constructing an obesity mouse model at high altitude and further treating the mice with anthocyanins,the expression of ADAM17 gene closely related to the occurrence of obesity and key proteins of PI3K/Akt signaling pathway were detected combined with gene sequencing technology and network pharmacology results.The results showed that ADAM17 promoted obesity by inhibiting PI3K/Akt signaling pathway.Anthocyanins activate PI3K/Akt signaling pathway by down-regulating ADAM17 and promote white fat Browning and mitochondrial function at high altitude.In conclusion,it was found that high altitude environment can improve the body weight,serum lipid and inflammatory factors levels,and promote the Browning of white fat in obese mice.Anthocyanins promote white fat Browning in obese mice at high altitude by downregulating the expression of ADAM17 in i WAT and activating PI3K-Akt signal.These results provide a theoretical basis for regulating lipid metabolism and breeding of livestock and poultry,and enrich the research basis for the prevention and treatment of human obesity and its related metabolic syndrome.