Effect Of DNA Methylation On Adipose Deposition In Pigs

Adipose deposition is an important indicator for evaluating the animal meat quality,as an economic animal closely related to people's lives,pigs are considered to be a good model for studying the mechanism of adipose deposition.Previous studies have shown that DNA methylation can regulate the adipose deposition process in animals,but its specific regulation mechanism is still unclear.This article aims to explore the regulation of DNA methylation on pig adipose deposition,and to provide a reference for elucidating adipose deposition mechanism and improving animal meat quality.In order to determine the effect of DNA methylation on the proliferation and differentiation of porcine progenitor adipocytes,this experiment used subcutaneous adipose tissue in the back and neck of 1 day old piglets,and isolated the primary primordial adipocyte by collagenase digestion.MTT assay was used to detect the proliferation of pre-adipocytes after adding 5-Aza-dc.Oil red O staining was used to observe the formation of lipid droplets in mature adipocytes after adding 5-Aza-dc.Expression levels of DNA methyltransferase and adipocyte differentiation-related genes during adipocyte proliferation and differentiation was detected by qRT-PCR.Then,taking Jin Fen White pig as the research object,the relationship between the DNA methylation degree of adipose tissue and the backfat,intramuscular fat,total cholesterol,triglyceride,hormone-sensitive lipase and lipoprotein lipase in serum were analyzed.The fat pigs and lean pigs were used as the research object,the DNA methylation degree and mRNA expression level of the adipose deposition marker genes were determined by BSP and qRT-PCR,exploring the effect of DNA methylation degree on related gene expression and adipose deposition process.The experiment results showed that:Adding methylation inhibitor?5-Aza-dc?can significantly increase the proliferation rate of porcine preadipocytes;8 ?mo L/L 5-Aza-dc effect 24 h is best to cell proliferation rate.DNMT1 may plays a major role in the process of 5-Aza-dc promoting the proliferation of preadipocytes.Adding 5-Aza-dc can significantly increase the expression of C/EBP_? and other adipose deposition genes in pig preadipocytes,increase lipid droplet formation.The Lower the degree of DNA methylation of adipose tissue,the thicker the backfat of the pig.The degree of DNA methylation was significantly negatively correlated with the backfat.DNA methylation could affect the formation of adipose tissue in the back of pigs.In adipose tissue of pig,the higher the degree of DNA methylation of related genes,the lower the expression level.The expression levels of PPAR?,Leptin and FABP₄ in pig adipose deposition are inversely proportional to the degree of DNA methylation.The expression levels and the degree of DNA methylation of PPAR?,Leptin and FABP₄ are inversely proportional.In conclusion,DNA methylation affect the adipose deposition of pig;The difference in adipose deposition ability between the Ma Shen pig and Large White pigs may be related to the regulation of DNA methylation of related genes.