从江香猪肌内前体脂肪细胞分化过程中相关基因的表达研究

doi:10.16431/j.cnki.1671-7236.2018.09.019

Abstract

Abstract:

In order to investigate the expression of the relate genes in the differentiation of intramuscular preadipocyte cells in Congjiang Xiang pig,the longissimus dorsi was collected from the 3-day-old Congjiang Xiang pig,type Ⅱ collagenase was adopted to digest the cells,and the intramuscular preadipocyte cells were separated and cultured,and then the morphology were observed.After induction,the intramuscular preadipocyte cells were identified by Oil red O staining method.Real-time PCR method was used to detect the expression levels of pyruvate dehydrogenase kinase 4(PDK4),fibroblast growth factor 10(FGF10),adiponectin (ADIPOQ),lipoprotein lipase(LPL),CCAAT/enhancer-binding protein α (C/EBPα),fatty acid synthase (FAS),adipocyte fatty acid binding protein 4 (FABP4) and protein kinase B (AKT2) at 0,24,48,72 and 144 h of cell differentiation,0 h was selected as control group.The results showed that the primary culture of intramuscular preadipocyte cells were started adherent in about 5 h,the shapes of adherent cells were round and transparent.After sub-culturing,the cell morphology was uniform,the cells showed red by Oil red O staining after induced culture.Real-time PCR results showed that the mRNA expression levels of PDK4,ADIPOQ,C/EBPα,FAS,FABP4 and AKT2 genes were all highly expressed at 48 h,which were extremely significantly higher than that in other stages (P<0.01);The mRNA expression level of FGF10 gene was higher in the induction of 24 and 48 h;The mRNA expression level of LPL gene at 72 h was extremely significantly higher than control group (P<0.01);The mRNA expression levels of PDK4,ADIPOQ and FGF10 genes at 144 h were extremely significantly lower than control group (P<0.01);The mRNA expression levels of C/EBPα gene at 144 h was significantly higher than control group (P<0.05);The mRNA expression levels of FAS gene at 144 h was significantly lower than control group (P<0.05);The mRNA expression levels of AKT2 and LPL genes at 144 h were not significantly different from that in control group (P>0.05).In this study,we successfully cultured the intramuscular preadipocyte cells,and the expression of adipose-related genes in different induction stages were detected,which provided the foundation for further study on lipid metabolism and deposition of Congjiang Xiang pig.

Key words: Congjiang Xiang pig; intramuscular preadipocyte cell; Real-time PCR; induction culture

CLC Number:

S813.3

XU Min, XU Houqiang, CHEN Wei, YANG Yang. Study on Expression of the Relate Genes Involved in Intramuscular Preadipocyte Differentiation in Congjiang Xiang Pig[J]. , 2018, 45(9): 2492-2499.

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Study on Expression of the Relate Genes Involved in Intramuscular Preadipocyte Differentiation in Congjiang Xiang Pig

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