血清对猪前脂肪细胞分化过程中聚脂相关基因表达的影响

Effects of Serum on Expression Models of the Genes Involved in Adipogenesis during Porcine Preadipocyte Differentiation

为了探讨血清对猪前脂肪细胞诱导分化的影响,筛选更优的诱导方法。采用胶原酶消化法分离猪皮下前脂肪细胞,用含50 nmol.L-1胰岛素、100 nmol.L-1地塞米松、0.25 mmol.L-13-异丁基-1-甲基黄嘌呤、100nmol.L-1罗格列酮及添加(对照组)或不添加(试验组)10%FBS的分化培养液1和2对前脂肪细胞进行诱导分化,借助实时定量RT-PCR方法检测了细胞分化过程中聚脂相关基因PPARα、C/EBPα、FASN、ACOX1、GPAT和ENPP2的表达模式。结果显示:血清对PPARγ和FABP4两基因的表达有极显著的上调作用(P<0.01),而对其他6个基因的表达有极显著的下调作用(P<0.01)。试验组中FASN、GPAT和ACOX1 3个基因诱导分化各时间点的综合表达量均极显著高于对照组(P<0.01),3基因表达量变化趋势间均达到极显著相关(P<0.01)。试验组中PPARα、PPARγ、C/EBPα和ACOX1 4个基因的基因表达量变化趋势间均达到显著或极显著正相关。研究表明:前脂肪细胞分化过程中,细胞内脂肪酸的生物合成和β氧化均在发生,细胞内脂肪含量积聚的快慢取决于2个途径力量的对比;PPARα、PPARγ、C/EBPα和ACOX1 4个基因间存在极强的协同表达现象;在细胞内脂肪积聚快慢上,含血清的分化培养液1要优于不含血清的分化培养液2。

To investigate the effects of serum on the preadipocyte during the process of induced differentiation and describe the expression patterns of genes related to adipogenesis in porcine,preadipocytes were isolated from subcutaneous adipose tissue of two days old piglets（Large White × Rongchang pigs） by using collagenase-digestion approach.The preadipocytes were cultured in different media supplemented with 50 nmol·L-1 insulin,100 nmol·L-1 dexamethasone,0.25 mmol·L-1 1-methyl-3-isobutylxanthine and 100 nmol·L-1 rosiglitazone.In addition,the different effects between the two methods were also been compared,which included the effects of serum（control group） and serum-free（experimental group） on differentiation of preadipocytes,respectively.Total RNA were extracted from the cells and the mRNA expression level of genes involved in adipogenesis were measured by real-time quantitative RT-PCR method.The results indicated that the mRNA expression levels of PPARα,C/EBPα,FASN,ACOX1,GPAT and ENPP2 were significantly down-regulated（P〈0.01） under the condition of serum added,in contrast,the mRNA expression level of PPARγ and FABP4 were significantly up-regulated（P〈0.01）.The mRNA expression level of FASN,GPAT and ACOX1 in the experimental group were higher than that in the contrast group at P〈0.01 level.Strikingly,the significant positive correlations among the mRNA expression levels of PPARα,PPARγ,C/EBPα and ACOX1 were observed in the experimental group.The results indicated that the serum has a fairly strong inhibitory discoloring effect on porcine preadipocyte during the process of induced differentiation.PPARα,PPARγ,C/EBPα and ACOX1 have the similar expression patterns.The serum is essential to accelerate the deposition of intra-cellular fat.