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

摘要/Abstract

摘要：

试验旨在探讨从江香猪肌内前体脂肪细胞分化过程中相关基因的表达。采集3日龄从江香猪背最长肌，采用Ⅱ型胶原酶消化法分离肌内前体脂肪细胞，进行原代和传代培养，并对其进行形态学观察。诱导培养后，利用油红O染色法对其进行鉴定。采用实时荧光定量PCR方法检测细胞诱导分化0、24、48、72和144 h时脂肪相关基因丙酮酸脱氢酶激4（PDK4）、成纤维细胞生长因子10（FGF10）、脂联素（ADIPOQ）、脂肪酸合成酶（FAS）、脂蛋白脂酶（LPL）、CCAAT增强子结合蛋白α（C/EBPα）、脂肪细胞脂肪酸结合蛋白4（FABP4）、蛋白激酶B（AKT2）的表达，选择诱导0 h作为对照组。结果显示，分离的肌内前体脂肪细胞5 h开始贴壁，贴壁的细胞呈圆形，胞体透明，经传代后，细胞形态均一，经诱导培养后，油红染色呈红色。实时荧光定量PCR结果显示，PDK4、ADIPOQ、C/EBPα、FAS、FABP4和AKT2基因mRNA表达水平在诱导48 h时均呈现较高表达，极显著高于其余各阶段（P<0.01）；FGF10基因mRNA表达水平在诱导24和48 h时均较高；LPL基因mRNA表达水平在诱导72 h时极显著高于对照组（P<0.01），之后明显下降；PDK4、ADIPOQ和FGF10基因mRNA表达水平在诱导144 h时均极显著低于对照组（P<0.01）；C/EBPα基因mRNA表达水平在诱导144 h时显著高于对照组（P<0.05）；FAS基因mRNA表达水平在诱导144 h时显著低于对照组（P<0.05）；AKT2和LPL基因mRNA表达水平在诱导144 h时与对照组差异不显著（P>0.05）。本试验成功培养了从江香猪肌内前体脂肪细胞，并检测了不同诱导阶段脂肪相关基因的表达情况，为进一步研究从江香猪脂肪代谢和沉积提供参考依据。

关键词: 从江香猪; 肌内前体脂肪细胞; 实时荧光定量PCR; 诱导培养

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

中图分类号:

S813.3

徐敏, 许厚强, 陈伟, 杨洋. 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[J]. 《中国畜牧兽医》, 2018, 45(9): 2492-2499.

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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