Study On Culture Of Porcine Adipocytes In Vitro And Lipogenesis Induced By Glucose And Insulin

Adipose tissue is not only a triglyceride repositories, but also an active endocrinal organ in energy metabolism of whole body. Adipogenesis was the process that adipocytes increased both amount and volume into mature adipocyte and triglyceride synthesis. Adipogenesis is influenced with many factors, such as nutrients and hormones. Carbohydrate is the main energy source and lipogenic substrate for animals, and it and hormones regulate adipogenesis together. After intaking carbohydrate, the secretion of insulin of pancreatic (3-cell was increase, and transcriptional expression of gene of glycolysis and fatty acid synthesis were regulated. Therefore, study on transcription factor and lipogenic-related genes of lipogenesis induced by glucose and insulin, that is important to elucidate the molecular mechanism of adipogenesis regulation and control of adipogenesis.Use neck and scapular subcutaneous adipose tissue got from7-day old piglet as experimental materials, preadipocytes were isolated and cultured by collagenase digestion method。Choose the best solution of porcine preadipocyte differentiation to mature adipocyte by use of oil red O staining extraction. We investigate effect of glucose and insulin on lipogenisis and transcriptional expressions of lipogenic-related genes in mature adipocytes in vitro, to explore mechanisms of glucose and insulin regulation of lipogenesis. The main results were as follows:1.The adipocytes were derived from porcine subcutaneous adipose tissue and can be sub-cultured and expanded stably, but the ability of self-differentiation is weak.2.By oil red O staining extraction, several methods used to establish the best method of the porcine preadipocyte differentiation to the adipocyte. The best method was DMEM/F12+10%FBS.+100nmol/L INS+100nmol/L DEX+0.05mmol/L IBMX+100nmol/L RSG. Thus, in appropriate vitro condition, porcine preadipocyte could be induced to mature adipocyte, and different regulatory pathway synergistically promoted cell differentiation.3.Lipogenesis and the mRNA levels of lipogenic genes FAS and ACC1and ChREBP were increased by glucose treatments and this stimulation of glouse was dose-dependent within concentration of20mmol/L in primary cultured mature adipocytes.but glucose had no effect on the expression of transcription factors SREBP-1c.4Lipogenesis was not altered by insulin treatments under culture condition of low glueose(5mmol/L).Lipogenisis and the mRNA levels of lipogenic genes FAS and and SREBP-lc were significantly promoted by insulin treatmemts in high glucose media(15mmol/L) in primary cultured mature adipocytes, but insulin had no effect on the transcription expression of ChREBP and ACC1. The results suggest that, glucose maybe induce lipogenic enzyme genes expression and lipogenic by the transcription factor ChREBP in primary cultured porcine adipocytes, and insulin maybe induce lipogenic enzyme genes expression and lipogenic by the transcription factor SREBP-lc in primary cultured porcine adipocytes, but further research is needed to prove.