绵羊脂肪细胞去分化和诱导再分化特性及关键基因表达规律研究  

A Study of Characteristics and Key Genes Expression Patterns of Sheep Adipocytes in Dedifferentiation and Induced Redifferentiation State

作　　者：张伟[1] 郭庆河[1] 崔炳灿[1] 高莉[1] 宁程程 牛嘉 王世银[1] 吕刚 ZHANG Wei;GUO Qinghe;CUI Bingcan;GAO Li;NING Chengcheng;NIU Jia;WANG Shiyin;LYU Gang(Key Laboratory of Livestock and Poultry Healthy Breeding Technology in Northwest China,Ministry of Agriculture and Rural Affairs,Xinjiang Agricultural Vocational Technical College,Changji 831100,China;Western Research Institute,Chinese Academy of Agricultural Science,Changji 831100,China;Xinjiang Taikun Group Co.,Ltd.,Changji 831100,China)

机构地区：[1]新疆农业职业技术学院,农业农村部西北畜禽健康养殖技术重点实验室,昌吉831100 [2]中国农业科学院西部农业研究中心,昌吉831100 [3]新疆泰昆集团有限责任公司,昌吉831100

出　　处：《动物营养学报》2024年第7期4638-4653,共16页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：新疆维吾尔自治区自然科学基金杰出青年科学基金项目(2022D01E18);新疆维吾尔自治区自然科学基金面上项目(2022D01A92);新疆维吾尔自治区现代农业羊产业技术体系。

摘　　要：本试验旨在初步探究绵羊脂肪细胞去分化机制和诱导再分化能力差异。分别使用经优化的“天花板”培养法研究不同月龄和组织来源脂肪细胞去分化能力,及去分化过程中细胞形态和关键基因的表达,并对不同组织来源去分化脂肪(DFAT)细胞诱导成脂分化能力进行比较。结果表明:若不使用不同孔径的细胞筛筛选脂肪细胞大小,不同月龄相同来源的脂肪细胞完成去分化的时间无显著差异(P>0.05);相同月龄绵羊的尾部脂肪细胞和肾周脂肪细胞完成去分化的时间无显著差异(P>0.05),但均极显著长于肌内脂肪细胞(P<0.01)。若将脂肪细胞筛选为不同大小的细胞组,则不论是尾部脂肪细胞还是肌内脂肪细胞,直径<300目细胞筛孔径的细胞完成去分化的时间均极显著短于直径>300目细胞筛孔径的细胞(P<0.01),直径>200目细胞筛孔径的尾部脂肪细胞完成去分化的时间又极显著长于直径200~300目细胞筛孔径的细胞(P<0.01),证明脂肪细胞完成去分化的时间与其细胞大小有关,而与其来源无明显相关性。脂肪细胞启动去分化后,细胞中的大脂滴逐步裂解为小脂滴,调控脂质合成和分解的基因均出现极显著上调(P<0.01),同时在细胞培养基中检测到甘油三酯含量出现极显著上升(P<0.01),而游离脂肪酸的含量虽有显著上升(P<0.05),但在整个去分化过程中基本保持稳定,推断脂肪细胞在去分化过程中主要通过直接排出脂滴完成细胞中脂质的清除。随着脂肪细胞的去分化,脂肪细胞祖细胞和前体脂肪细胞标志基因以及干细胞标志基因均出现极显著上调(P<0.01),说明脂肪细胞逆转了细胞命运决定而重新获得了干细胞特性。不同组织来源的DFAT细胞诱导成脂分化能力无显著差异(P>0.05)。综上所述,绵羊脂肪细胞大小与其去分化能力极显著相关,但与其组织来源和年龄无明显相关性。DFAT细胞主要通过排出脂滴完The aim of this experiment was to investigate the differences in dedifferentiation mechanisms and induction redifferentiation abilities of sheep adipocytes.The optimized‘ceiling’culture method was utilized to examine the dedifferentiation ability of adipocytes derived from different months of age and tissue sources,as well as the cell morphology and expression of key genes during dedifferentiation.Additionally,the induced lipogenic redifferentiation ability of dedifferentiated fat(DFAT)cells from different tissue sources were compared.The results showed as follows:if adipocytes were not screened using cell screens with different mesh sizes,there were no significant differences in the dedifferentiation time among adipocytes originating from same tissue but different months of age(P>0.05);there were no significant differences in dedifferentiation time between tail adipocytes and perirenal adipocytes within the same month of age(P>0.05),but both significantly longer than intramuscular adipocytes(P<0.01).If adipocytes were screened into cell groups of different sizes,the dedifferentiation time for both tail adipocytes and intramuscular adipocytes with a diameter less than 300 mesh cell sieve apertures was significantly shorter than that with a diameter greater than 300 mesh cell sieve apertures(P<0.01),the dedifferentiation time for tail adipocytes with a diameter larger than 200 mesh cell sieve apertures was significantly longer than that with diameters ranging between 200 and 300 mesh cell sieve apertures(P<0.01).So,it was confirmed that the time of adipocyte dedifferentiation was related to their size rather than their tissue source.After the dedifferentiation of adipocytes,large lipid droplets in cells gradually fragmented into smaller ones,accompanied by significant upregulation of genes related to lipid synthesis and decomposition processes(P<0.01),and significant increase in triglyceride content detected in the cell medium(P<0.01),while the free fatty acid content also showed a significant increase but kee

关 键 词：绵羊 脂肪细胞 脂质 脂滴 去分化 DFAT 诱导分化 

分 类 号：S811.3[农业科学—畜牧学]