Pseudo-targeted metabolomic analysis of Chinese Herbal Medicines Cangfu Daotan Decoction to improve in vitro fertilization outcomes of patients with Polycystic Ovary Syndrome

Abstract

Objective To explore the mechanism of Cangfu Daotan Decoction (CDD) about improving the in vitro fertilization (IVF) outcomes of patients with polycystic ovary syndrome (PCOS) based on pseudo-targeted metabolomics. Methods We included 95 subjects of infertility patients participating in in vitro fertilization (IVF) protocols, including 65 patients in PCOS intervened with or without CDD (PCOS+CDD Group, n = 33; PCOS Group, n = 32) and 30 healthy women (Control Group, n = 30). We compared the clinical outcomes among the three groups, Pseudo-targeted metabolomics method was used to analyze follicular fluid samples on the day of oocytes retrieval and to further explore related metabolites and metabolic pathways. Based on ultra-performance liquid chromatography-mass spectrometry (UPLC-MS)，Pseudo-targeted metabolomics was implemented to explore the related metabolites and metabolic pathways among three groups through professional MetaboAnalyst analysis software. The clinical outcomes were collected among the three groups. Follicular fluid samples on the day of oocytes retrieval were analyzed via pseudo-targeted metabolomics to further explore related metabolites and metabolic pathways. Results There were statistically significant differences among three groups’ outcome indexes, including the number of total retrieved oocytes, metaphase II oocytes, 2PN cleavage, transferable embryo, high-quality embryo fertilization rate and cleavage rate (P<0.05). Moreover, these differences before and after Chinese medicine intervention were statistically significant in cleavage rate (76.1% VS 88.3%) and blastogenesis rate (46.2% VS 80.6%) (P < 0.001). Compared with ones in PCOS group, seven metabolites in PCOS+CDD Group, namely Leucine, N-acetyltryptophan, 3-sulfopyruvic acid, uric acid, Acrylamide, creatine, lactate, were up-regulated. And there were others down-regulated substances: progesterone, phytosphingosine, decanoylcarnitine, dihomolinoleic acid, choline, phosphorylcholine, betaine, parahydroxybenzoate. The pathways containing steroid hormone biosynthesis, glycine, serine and threonine metabolism, valine, leucine and isoleucine biosynthesis, purine metabolism as well as pyruvate metabolism were regulated by CDD treatment. Conclusion CDD could improve the IVF outcomes of patients with polycystic ovary syndrome (PCOS) by increasing total retrieved oocytes, transferable embryos and high-quality embryos and enhancing retrieved oocytes rate, fertilization rate and clinical pregnancy rate. Follicular fluid (FF) metabolites of PCOS patients with spleen deficiency and phlegm-dampness syndrome involving lipids (aminools), organic acids (lactic acid) could be regulated by CDD. These differential metabolites may provide potential biomarkers for PCOS patients with spleen deficiency and phlegm-dampness syndrome. CDD can improve pyruvate metabolism and correct lipid metabolism disorders, which may be the possible mechanism of action for improving ART outcomes in PCOS patients.