鸡体细胞诱导重编程早期的糖代谢变化研究  

Changes of Glucose Metabolism in Early Stage of Chicken Somatic Cell Induced Reprogramming

作　　者：袁霞 王佳琪[2] 王美瑰 孙国兴 姜景译 石祥 张明 丁颖[1,2] 左其生[1,2] 张亚妮 李碧春 YUAN Xia;WANG Jiaqi;WANG Meigui;SUN Guoxing;JIANG Jingyi;SHI Xiang;ZHANG Ming;DING Ying;ZUO Qisheng;ZHANG Yani;LI Bichun(Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China,Yangzhou,Jiangsu 225009;College of Animal Science and Technology,Yangzhou University,Yangzhou,Jiangsu 225009)

机构地区：[1]教育部农业与农产品安全国际合作联合实验室,江苏扬州225009 [2]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《中国家禽》2022年第1期11-18,共8页China Poultry

基　　金：国家自然科学基金项目(31872341、31772582);国家重点研发计划(2017YFE0108000);扬州大学研究生科研创新计划项目(XKYCX19_123);大学生创新创业训练计划项目(201911117106Y)。

摘　　要：为研究鸡体细胞诱导重编程早期的糖代谢方式的变化,试验采用OCT4、SOX2、NANOG和LIN28A(OSNL)四因子诱导体系将鸡胚成纤维细胞(Chicken embryo fibroblasts,CEF)重编程为诱导多能干细胞(Induced pluripotent stem cells,iPS),并利用碱性磷酸酶染色、阶段特异性胚胎抗原1(Stage-specific embyronic antigen-1,SSEA-1)免疫荧光染色、体外诱导分化及多能性基因表达检测等对iPS进行鉴定。通过检测重编程过程中糖代谢相关基因表达及酶活性的变化,并对葡萄糖摄取量、乳酸产生量及线粒体膜电位检测等研究鸡体细胞诱导重编程早期的糖代谢变化。结果显示,鸡CEF诱导重编程形成的iPS呈碱性磷酸酶染色阳性,表达SSEA-1蛋白,体外分化形成类胚体且表达多能性标记基因。同时重编程过程中氧化磷酸化基因表达下调而糖酵解相关基因表达上调,糖酵解关键酶活性均增强,且iPS的葡萄糖吸收量及乳酸产生量增加,而线粒体膜电位则下降。结果表明,OSNL四因子体系将鸡CEF诱导重编程形成iPS的过程中,细胞的主要糖代谢方式从氧化磷酸化转变为糖酵解,而糖酵解的激活可能会进一步促进iPS的形成。In order to study the changes of glucose metabolism in early stage of chicken somatic cell induced reprogramming,chicken embryo fibroblasts(CEF)were reprogrammed into induced pluripotent stem cells(iPS)by OCT4,SOX2,NANOG and LIN28 A(OSNL)induction strategy.iPS was identified by alkaline phosphatase staining,stage-specific embyronic antigen-1(SSEA-1)immunofluorescence,in vitro induced differentiation and pluripotentgeneexpression detection.The expression of glucose metabolism related genes and enzyme activity,glucose uptake,lactate production and mitochondrial membrane potential were detected to study the changes of glucose metabolism in chicken somatic cells induced reprogramming.The results showed that iPS induced by CEF were positive for alkaline phosphatase staining,expressed SSEA-1 protein,differentiated into embryoids in vitro and expressed pluripotent marker gene.At the same time,during the reprogramming process,the expression of oxidative phosphorylation genes was down regulated,while the expression of glycolysis related genes was up-regulated.The activities of key enzymes in glycolysis were enhanced,and the glucose uptake and lactate production of iPS were increased,while the mitochondrial membrane potential was decreased.The results suggested that in the process of chicken CEF induced reprogramming to iPS by OSNL four factors system,the main mode of glucose metabolism changed from oxidative phosphorylation to glycolysis,and the activation of glycolysis may further promote the formation of iPS.

关 键 词：鸡 诱导重编程 诱导多能干细胞 糖代谢 糖酵解 

分 类 号：S831.2[农业科学—畜牧学]