泛酸及其下游脂肪酸氧化代谢在鸡ESCs向PGCs分化过程中的作用  

The Role of Pantothenic Acid and its Downstream Fatty Acid Oxidation Metabolism in the Differentiation Process of Chicken ESCs into PGCs

作　　者：赵梓多 武嵘峰 耿晴晴 程富富 王哲 吴一凡 张亚妮 ZHAO Ziduo;WU Rongfeng;GENG Qingqing;CHENG Fufu;WANG Zhe;WU Yifan;ZHANG Yani(Jiangsu Key Laboratory of Animal Genetic Breeding and Molecular Design,College of Animal Science and Technology,Yangzhou University,Yangzhou,Jiangsu 225009,China)

机构地区：[1]扬州大学动物科学技术学院/江苏省动物遗传繁育与分子设计重点实验室,江苏扬州225009

出　　处：《家畜生态学报》2024年第12期37-42,共6页Journal of Domestic Animal Ecology

基　　金：国家自然科学基金项目(32272857);国家重点研发计划(2021YFD120030508);江苏省研究生实践创新计划(SJCX23_1995)。

摘　　要：本文旨在探究差异代谢物泛酸(PA)和脂肪酸氧化(FAO)在鸡胚胎干细胞(ESCs)向原始生殖细胞(PGCs)分化过程中的作用,为从代谢角度解析PGCs形成机制提供新的理论基础。采用BMP4诱导体系,基于课题组前期对泛酸及其抑制剂(PZ)和脂肪酸氧化激活剂(BMS)/抑制剂(Per)的最适添加浓度,设置5个分组:Control组、PA+BMS组、PA+Per组、PZ+BMS组、PZ+Per组。通过观察细胞形态、qRT-PCR检测PGCs特异标记基因、间接免疫荧光和流式细胞分选等探究泛酸代谢及其下游的脂肪酸氧化在ESCs向PGCs分化过程中的作用。细胞形态观察结果显示,诱导至第6天,PZ+BMS组的类胚体数目显著升高(P<0.01),PA+Per组显著减少(P<0.05);qRT-PCR结果显示,CVH和C-KIT在PZ+BMS诱导组中的表达量极显著升高(P<0.01),而在PA+Per组中的表达量则显著降低(P<0.01);间接免疫荧光和流式细胞分选结果显示,PZ+BMS组DDX4阳性细胞率显著高于对照组(P<0.05),而PA+Per组DDX4阳性细胞率极显著低于对照组(P<0.01)。以上结果表明,抑制泛酸代谢并激活脂肪酸代谢能够促进PGCs的形成,该研究为利用代谢物完善PGCs的体外诱导培养体系提供新见解。This article aims to investigate the roles of differential metabolites pantothenic acid(PA)and fatty acid oxidation(FAO)in the differentiation of chicken embryonic stem cells(ESCs)into primordial germ cells(PGCs),and to provide a new theoretical basis for analyzing the mechanism of PGCs formation from a metabolic perspective.Using BMP4 induction system,five groups were established based on the optimal concentrations of pantothenic acid(PA)and its inhibitor(PZ)and fatty acid oxidation activator(BMS)/inhibitor(Per)in previous studies of the research group:Control,PA+BMS,PA+Per,PZ+BMS,and PZ+Per.Cell morphology was observed,PGCs-specific marker genes were detected by qRT-PCR,and indirect immunofluorescence and flow cytometry were employed to explore the effects of pantothenate metabolism and its downstream fatty acid oxidation during ESCs to PGCs differentiation.Cell morphology observations at day 6 of induction showed a significant increase in embryoid bodies in the PZ+BMS group(P<0.01)and a significant decrease in the PA+Per group(P<0.05).qRT-PCR results indicated a significant increase in CVH and C-KIT expression in the PZ+BMS-induced group(P<0.01),while their expression was significantly reduced in the PA+Per group(P<0.01).Indirect immunofluorescence and flow cytometric sorting results demonstrated a significantly higher proportion of DDX4-positive cells in the PZ+BMS group compared to the control(P<0.05),whereas the PA+Per group showed a significantly lower proportion(P<0.01).These findings suggest that inhibiting pantothenic acid metabolism and activating fatty acid metabolism can promote PGCs formation.This study provides new insights into the optimization of in vitro induction culture system of PGCs using metabolites.

关 键 词：泛酸 脂肪酸氧化 PGCS 

分 类 号：S831.2[农业科学—畜牧学]