代谢组学分析疤痕憩室合并宫腔积液IVF/ICSI患者新鲜胚胎移植的妊娠影响

To Analyze the Factors of Pregnancy Outcome in IVF/ICSI Patients with Previous Cesarean Scar Defect and Endometrial Cavity Fluid Based on Metabonomics

目的:本文通过在体外受精(IVF)或卵胞浆内单精子显微注射(ICSI)新鲜周期移植前,对有剖宫产疤痕憩室(PCSD)合并宫腔积液的患者抽取宫腔积液,将抽出的宫腔积液采用代谢组学方法分析代谢差异物,从而探讨宫腔积液对临床妊娠影响的分子机制。方法:将符合入组条件的患者宫腔积液样本根据移植后是否临床妊娠分为两组,临床妊娠组G1(n=7)和非临床妊娠组G2(n=6),进行液相色谱串联质谱技术检测,对数据进行统计学和生物信息学分析。结果:两组比较共鉴定出31种代谢产物存在差异,其中临床妊娠组上调表达代谢物30种(FC≥2.0,P<0.05),下调表达代谢物1种(FC≤0.5,P<0.05)。对代谢物进行京都基因与基因组百科全书(KEGG)富集分析,结果差异代谢物在亚精胺和精胺生物合成、嘌呤代谢、精氨酸和脯氨酸代谢等代谢通路中富集明显,其中亚精胺和精胺生物合成通路富集最显著,涉及的差异代谢物为鸟氨酸、亚精胺和精胺。结论:剖宫产疤痕憩室新鲜周期移植患者宫腔积液可能存在氨基酸代谢,乙酰辅酶合成等多条生物代谢通路的异常变化。核苷酸、氨基酸等被影响会间接或直接影响到三羧酸循环、糖酵解等代谢过程,这些生物学过程的改变可能影响到胚胎的种植。

Objective:To explore the molecular mechanism of endometrial cavity fluid’s(ECF)metabolites, which were extracted from patients with cesarean scar diverticulum(PCSD)and Endometrial Cavity Fluid(ECF),on the clinical pregnancy outcome by metabonomics.ECF collection occurs before the patient’s external in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)fresh embryo transfer cycle.Methods:The ECF of patients who met the enrollment conditions were divided into two groups according to the clinical pregnancy outcome, the clinical pregnancy group G1(n=7)and the non-clinical pregnancy group G2(n=6).Each group of samples was tested by liquid chromatography-tandem mass spectrometry, and the data was analyzed by statistics and bio-informatics.Results:A total of 31 metabolites were identified between the two groups.Among them, 30 metabolites were up-regulated in the clinical pregnancy group(FC≥2.0,P<0.05),and 1 metabolite was down-regulated(FC≤0.5,P<0.05).The Kyoto Encyclopedia of Genes and Genomics(KEGG)enrichment analysis of metabolites showed that differential metabolites were significantly enriched in the metabolic pathways of spermidine and spermine biosynthesis, purine metabolism, arginine and proline metabolism, etc.Among them, spermidine and spermine biosynthetic pathways are the most enriched, and the different metabolites involved are ornithine, spermidine and spermine.Conclusion:There may be abnormal changes in multiple biological metabolic pathways such as amino acid metabolism and acetyl-coenzyme synthesis in the endometrial cavity fluid of patients with previous cesarean scar defect undergoing fresh embryo transfer cycle.Nucleotides and amino acids will indirectly or directly affect metabolic processes such as the tricarboxylic acid cycle and glycolysis.Changes in these biological processes may affect the implantation of embryos.