鸡肝细胞中C/EBPα基因的敲低及对糖脂代谢相关基因表达的影响

Knock-down of C/EBPαGene Expression in Chicken Hepatocytes and Its Effect on the Expression of Lipid Metabolism Related Genes

C/EBPα即CCAAT/增强子结合蛋白α基因,其编码的CCAAT/增强子结合蛋白是重要的转录调控因子,在细胞的分化和代谢过程中发挥着重要的调节作用。试验利用RNAi来敲低鸡LMH细胞系中C/EBPα基因的表达,以研究C/EBPα基因影响脂肪代谢的机制。根据鸡C/EBPα基因的mRNA序列,设计3对siRNA,用RNAi干扰技术来敲低鸡LMH细胞中的C/EBPα基因表达,然后用荧光定量PCR检测C/EBPα基因的表达情况,发现其中的C/EBPα-siRNA-161能够显著降低C/EBPα基因的表达;并将C/EBPα-siRNA-161转染LHM细胞后48 h后,利用荧光定量PCR检测LMH细胞中糖脂代谢相关基因(PCK1、SCD、GLUT2、PPARγ、INS、G6PC、IGF1、IGF2、INSR、STAT3)的表达,发现在LMH细胞中PCK1、G6PC、GLUT2基因的mRNA表达量下降,结果表明C/EBPα基因能影响糖代谢相关基因G6PC、GLUT2、甘油异生相关基因PCK1的表达,证实了C/EBPα基因对鸡肝细胞中的糖脂代谢具有调控作用,通过调控C/EBPα基因的表达将影响糖吸收及甘油异生。

C/EBPα,CCAAT/enhancer binding proteinαgene,encodes CCAAT/enhancer binding protein,which is an important transcriptional regulator and plays an important regulatory role in the differentiation of adipocytes and metabolism.The purpose of this study was to knock down the expression of C/EBPαgene by RNAi in chicken LMH cell line and to investigate the mechanism of C/EBPαgene affecting lipid metabolism.Based on chicken C/EBPαmRNA sequence,3 siRNAs was designed to knock down the expression of C/EBPαgene in chicken hepatocytes LMH,and then the expression of C/EBPαgene was detected by fluorescence quantitative PCR.The results showed C/EBPα-siRNA-161 could significantly decrease the expression of C/EBPαgene.Further,C/EBPα-siRNA-161 was transferred into LMH cells for 48 h,the expression of PCK1,SCD,GLUT2,PPARγ,INS,G6 PC,IGF1,IGF2,INSR and STAT3 were detected by fluorescence quantitative PCR to observe the glycolipid regulated by C/EBPαgene.These results showed that the mRNA expression of PCK1,G6 PC and GLUT2 gene were decreased in LMH cells,while some studies showed that G6 PC and GLUT2 were related to glucose metabolism in liver,and PCK1 was involved in glycerol heterophysis,suggesting that C/EBPαgene can regulate glycolipid metabolism in chicken hepatocytes,and the expression of C/EBPαgene can affect glucose absorption and glycerol heterogenesis.