Bamei Pig Intramuscular And Subcutaneous Preadipocytes In Vitro Culture And Different Expression Of Lipogenic Genes

Swine as the main land animals the adipose tissue in the body fat distribution and deposition is the key factor affect the carcass quality and flavor of meat, subcutaneous fat affect carcass quality and intramuscular fat is closely related to the quality of muscle. Many studies have shown that lipid metabolism of intramuscular adipose tissue and subcutaneous adipose tissue is different, that the proliferation differentiation and metabolism of intramuscular and subcutaneous adipocytes have different regulation mechanism. Differential mRNA expression profiles of Bamei pig intramuscular preadipocytes compared with subcutaneous preadipocytes during differentiation,that is important significant to elucidate the molecular mechanism of adipogenesis regulation and control the formation of adipogenesis. Moreover,Bamei pig as local pig breeds its lipogenic-related genes of lipogenesis in different tissue have bigger difference, its regulatory mechanism is very complex, suggests this differences with fat metabolism and fat deposition has certain relations.Use3day old Bamei piglet as experimental animal,By collagenase digestion method from subcutaneous fatty tissue and longissimus dorsi got intramuscular and subcutaneous preadipocytes.Choose MTT colorimetric method and cell count pigskin to study the proliferation characteristics of the subcutaneous and intramuscular preadipocyte. Oil red O extraction measured the fat content in the induction differentiation process.Real-time quantitative PCR study the subcutaneous and intramuscular adipocytes of lipogenic-related genes of lipogenesis including FAS、 ACC1and transcription factors ChREBP and SREBP-lc expression differences,At the same time, By real-time quantitative PCR research Subcutaneous adipose,Small intestine,Liver,longissimus dorsi,Kidney,Heart and small intestine tissue of Bamei pig, excellent characters of genes for the local pig breeds also use provides the basis.The main results were as follows:1.The intramuscular and subcutaneous preadipocytes were derived from Bamei pig subcutaneous adipose tissue and longissimus dorsi tissue and can be sub-cultured and expanded,Oil red O staining extraction method of determination of fat content in the cell, The results showed that intracellular triglyceride concentrations of intramuscular preadipocytes increased more dramatically than that of subcutaneous preadipocytes during cell differentiation,but after the induction8d, subcutaneous adipocytes significantly higher lipid content of intramuscular adipocytes.2. Applying real-time quantification PCR technology to study the expression change and lipid levels that bamei pig intramuscular adipocytes and subcutaneous adipocytes of lipogenic-related genes of lipogenesis,the ChREBP and SREBP-lc mRNA expression level of subcutaneous adipocytes was significantly higher than that of intramuscular adipocytes (P<0.01), at the early of induction subcutaneous and intramuscular adipocytes of FAS and ACC1mRNA expression difference was not significant,after induced,subcutaneous adipocytes of mRNA expression level was significantly higher than that of intramuscular adipocytes (P<0.01).3.Glucose stimulated fatty acid biosynthesis and lipogenisis through inducing mRNA expressions of lipogenic-related genes FAS,ACC1,SCD and ChREBP in primary cultured intramuscular adipocytes and subcutaneous adipocytes, but do not increased SREBP-lc mRNA level. ChREBP mRNA expression in the intramuscular adipocytes abundance than subcutaneous adipocytes.4. Apply ing real-time quantification PCR technology to study the gene mRNA expression of bamei pigs in different tissue, the results show that the ChREBP, FAS and ACC1are expressed in various tissues, and extremely significant differences between different groups (P<0.01); the expression of SREBP-1c is very low in the liver, but in the subcutaneous fat, longissimus dorsi and kidney has high abundance level expression.