生殖医学杂志2025，Vol.34Issue(9)：1219-1228,10.DOI:10.3969/j.issn.1004-3845.2025.09.010
基于高效液相色谱-质谱技术分析不同发育潜能胚胎D5培养液的代谢差异
Analysis of metabolic differences in day 5 culture media of embryos with varying developmental potential by high performance liquid chromatography-tandem mass spectrometry
摘要
Abstract
Objective:To analyze the changes in metabolic profiles of D5 spent culture medium(SCM)from embryos with different developmental potentials,and to explore the metabolites and metabolic pathways that played important roles in blastocyst formation. Methods:Patients who underwent in vitro fertilization and embryo transfer(IVF-ET)treatment at the Reproductive Medicine Center of Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from November 13 to December 5,2023 were recruited.The D5 SCM from embryos were collected.The embryos were divided into D5 high-quality blastocyst group,D5 non high-quality blastocyst group and D5 non blastocyst formation group according to the blastocyst formation.The culture media of each group were subjected to non-targeted metabolomic analysis using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS).The differential metabolites were screened and the bioinformatics analysis was conducted on them. Results:A total of 118 culture media were included in this study for analysis,with a D5 blastocyst formation rate of 46.88％and a high-quality D5 blastocyst formation rate of 32.81％.The metabolic profiles of D5 culture medium exhibit significant variations depending on the developmental potential of embryos.A total of 80 differential metabolites were screened between the D5 blastocyst formation group(including D5 high-quality blastocyst group and D5 non high-quality blastocyst group)and the D5 non blastocyst formation group(P＜0.05),while there were 44 differential metabolites between the D5 high-quality blastocyst group and the D5 non high-quality blastocyst group(P＜0.05).Among the differential metabolites,there were significant differences(P＜0.05)in the differential metabolites of L-4-chlorotryptophan and pantothenic acid between the D5 blastocyst formation group and the D5 non blastocyst formation group,as well as between the D5 high-quality blastocyst group and the D5 non high-quality blastocyst group.KEGG pathway enrichment analysis revealed that the differential metabolites between the D5 blastocyst formation group and the D5 non blastocyst formation group were significantly enriched in pathways related to central carbon metabolism in cancer,glucagon signaling,glyoxylate and dicarboxylate metabolism,linoleic acid metabolism,the pentose phosphate pathway,and the biosynthesis of cofactors(P＜0.05).The differential metabolites between the D5 high-quality blastocyst group and the D5 non high-quality blastocyst group were primarily enriched in pathways associated with glyoxylate and dicarboxylate metabolism,vitamin digestion and absorption,glycerophospholipid metabolism,and the biosynthesis of cofactors(P＜0.05). Conclusions:There were significant differences in the D5 culture medium metabolomic profiles of embryos with different developmental potentials.Among the metabolites,L-4-chlorotryptophan and pantothenic acid had a significant impact on D5 blastocyst formation and blastocyst quality.The metabolic pathways of glyoxylate and dicarboxylate metabolism,as well as the biosynthesis of cofactors,played important roles in the process of blastocyst development.关键词
辅助生殖技术/胚胎发育潜能/胚胎培养液/代谢组学/高效液相色谱-质谱联用Key words
Assisted reproductive technology/Embryo development potential/Embryo culture medium/Metabolomics/High performance liquid chromatography-tandem mass spectrometry分类
医药卫生引用本文复制引用
陈雪花,陈焕华,黄贵婷,周红,舒金辉,汪彩珠..基于高效液相色谱-质谱技术分析不同发育潜能胚胎D5培养液的代谢差异[J].生殖医学杂志,2025,34(9):1219-1228,10.基金项目
国家自然科学基金(82460314) （82460314）
广西自然科学基金(2025GXNSFAA069468) （2025GXNSFAA069468）
广西生殖健康与出生缺陷防治重点实验室(广西壮族自治区妇幼保健院)开放课题资助(GXWCH-ZDKF-2023-05) （广西壮族自治区妇幼保健院）
广西中医药大学研究生教育创新计划项目(YCSY2025065) （YCSY2025065）
