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

发布时间：2018-09-05 08:41

【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(P0.05);LPL m RNA表达水平在分化早期各阶段无明显差异(P0.05),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(P0.01);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(P0.01);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(P0.01),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。
[Abstract]:In order to establish an in vitro culture model of (Sus scrofa) adipocyte precursor cells in Congjiang Xiang pig, the difference of expression of related genes in the differentiation of intramuscular and subcutaneous fat precursor cells of Congjiang Xiang pig was analyzed. The longissimus dorsi muscle and subcutaneous adipose tissue were collected and digested with collagenase to isolate adipose precursor cells from the two tissues. After induced differentiation, Using Q RT-PCR technique to detect adipose synthesis and decomposition genes during cell differentiation: transcription factor peroxisome proliferator activated receptor 纬 (peroxisome proliferator activated receptor 纬 PPAR 纬, lipoprotein lipase (lipoprotein lipase,LPL), fatty acid synthase (fatty acid synthase,FAS, acetyl coenzyme A carboxylase The temporal expression of acetyl-Co A carboxylase,ACC, triglyceride hydrolase (adipose triacylglycerol lipase,ATGL and hormone sensitive lipase (hormone-sensitive lipase,HSL). The in vitro culture model of fat precursor cells of Congjiang Xiang pig was successfully constructed. Q RT-PCR analysis showed that the expression of PPAR- 纬 FASACC m RNA in intramuscular adipocytes was higher at 72 h after differentiation. The expression level of LPLM RNA was significantly higher than that of 0 h (P0.05). There was no significant difference in the expression of LPLM RNA in the early stage of differentiation (P0.05), but it was significantly up-regulated at 144h. The expression level of HSLm RNA was significantly higher than that of other stages (P0.01). The expression level of PPAR 纬 gene in subcutaneous adipose progenitor cells was the highest at 72 h after differentiation, which was significantly higher than that in other stages (P0.01). The expression level was significantly higher than 0 h (P0.01), and then decreased. In vitro, the adipocyte differentiation related genes were active in the later stage of the differentiation of intramuscular adipocytes, while the adipocyte differentiation related genes were active in the early stage of subcutaneous adipose progenitor cell differentiation. The results showed that the formation of subcutaneous fat in Congjiang Xiang pig may be much earlier than that of intramuscular fat, which is of great significance to further study the mechanism of fat deposition in Congjiang Xiang pig.
【作者单位】：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;贵州大学动物科学学院;贵州省动物遗传育种与繁殖重点实验室;贵州大学生命科学学院;
【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)
【分类号】：S828

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