从江香猪肌内和皮下脂肪前体细胞分化过程中相关基因的表达  

The Expression of Related Genes on Intramuscular and Subcutaneous Preadipocytes During Differentiation in Congjiang Xiang Pig(Sus scrofa)

作　　者：孙成娟[1,2] 许厚强[1,3,2] 赵佳福[1,4] 陈伟[1,4] 周迪[1,4] 张青青[1,2] 

机构地区：[1]贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室,贵阳550025 [2]贵州大学动物科学学院,贵阳550025 [3]贵州省动物遗传育种与繁殖重点实验室,贵阳550025 [4]贵州大学生命科学学院,贵阳550025

出　　处：《农业生物技术学报》2017年第12期1979-1988,共10页Journal of Agricultural Biotechnology

基　　金：国家科技支撑计划(No.2015BAD03B02-3);贵州畜禽种质资源保存创新与利用(黔科合重大专项字(2013)6008);贵州大学研究生创新基金(研农2016021)

摘　　要：为建立从江香猪(Sus scrofa)脂肪前体细胞的体外培养模式,分析从江香猪肌内和皮下脂肪前体细胞分化过程中相关基因的表达差异。采集5日龄从江香猪背最长肌和皮下脂肪组织,利用胶原酶消化法,分别从此两种组织中分离获得脂肪前体细胞。经诱导分化后,采用q RT-PCR技术检测细胞分化过程中脂肪合成与分解基因:转录因子过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptorγ,PPARγ)、脂蛋白脂酶(lipoprotein lipase,LPL)、脂肪酸合成酶(fatty acid synthase,FAS)、乙酰辅酶A羧化酶(acetyl-Co A carboxylase,ACC)、甘油三酯水解酶(adipose triacylglycerol lipase,ATGL)、激素敏感脂酶(hormone-sensitive lipase,HSL)的时序表达情况。成功构建从江香猪脂肪前体细胞的体外培养模式。通过q RT-PCR分析显示,在肌内脂肪前体细胞中,PPARγ、FAS、ACC m RNA表达水平在整个分化早期72 h时均呈现较高表达,显著高于0 h的表达水平(P<0.05);LPL m RNA表达水平在分化早期各阶段无明显差异(P>0.05),但在144 h时出现显著的大幅度上调,极显著高于其他各阶段表达水平(P<0.01);ATGL、HSL m RNA表达水平在整个分化阶段均呈现"升高-降低-升高"的表达规律。在皮下脂肪前体细胞中,PPARγ基因在诱导分化72 h的表达水平最高,极显著高于其他几个阶段的表达水平(P<0.01);LPL、FAS、ACC、ATGL、HSL m RNA表达水平在整个分化早期48和72 h均呈现较高表达,极显著高于0 h的表达水平(P<0.01),之后均下降。在离体培养条件下,各脂肪细胞分化相关基因在肌内脂肪前体细胞的分化后期,表现较活跃;而各脂肪细胞分化相关基因在皮下脂肪前体细胞的分化早期,表现较活跃。研究结果表明从江香猪皮下脂肪的形成可能远远早于肌内脂肪,对进一步研究从江香猪脂肪沉积机制具有重要意义。In order to investigate the differential gene expression on intramuscular and subcutaneous preadipocytes during differentiation in Congjiang Xiang pig（Sus scrofa）, the fat precursors culture model in vitro of Congjiang Xiang pig was established. In this study, Congjiang Xiang pig in five days old was provided, then subcutaneous adipose tissue and the longissimus dorsi muscle was excised. We used collagenase to harvest preadipocytes from longissimus dorsi muscle and subcutaneous adipose tissue respectively. The two preadipocytes were induced differentiation by a series of hormone-induced complexes.The mRNA expression levels of gene involved in adipogenesis, such as peroxisome proliferator-activated receptorsγ（PPARγ）, lipoprotein lipase（LPL）, fatty acid synthase（FAS）, acetyl-Co A carboxylase（ACC）,adipose triglyceride lipase（ATGL） and Hormone sensitive lipase（HSL）, were compared by real-time PCR in the two preadipocytes. The result showed that the model of preadipocytes in vitro was successfully established from Congjiang pig. The mRNA expression levels of PPARγ, FAS and ACC in intramuscular preadipocytes were significantly a higher at 72 h after the addition of inducer, which a higher than the level in 0 h（P〈0.05）.The mRNA expression level of LPL was no significant difference in the various stages, but there was a significant up-regulation of LPL in 144 h after the addition of inducer, and the value was significantly higher than the other stages（P〈0.01）. The mRNA expression levels of ATGL and HSL showed the pattern of＂updown-up＂in the differentiation phase. In the subcutaneous preadipocytes, the mRNA expression level of PPARγ was the highest in 72 h after induction, and the value was significantly higher than in other stages（P〈0.01）. The mRNA expression levels of LPL, FAS, ACC, ATGL and HSL were all higher in 48 and 72 h, and the value was significantly higher than that in 0 h（P〈0.01）. In summary, this study showed that in intramuscular preadipocytes, the

关 键 词：从江香猪 肌内脂肪前体细胞 皮下脂肪前体细胞 时序表达 

分 类 号：S828.2[农业科学—畜牧学]