To Investigate The Pathogenesis Of PCOS With Phlegm-Dampness Based On The Concentration Of Phthalate Metabolites In Follicular Fluid

Objective:Based on targeted metabolomics,to explore the relationship between follicular fluid phthalates and their metabolites and the pathogenesis of polycystic ovary syndrome（PCOS）of phlegm-dampness type,in order to provide new insights into the diagnosis and prevention of PCOS and TCM syndrome differentiation and treatment.Methods:A prospective cohort study was conducted in 84 patients who underwent in vitro fertilization and embryo transfer（IVF-ET）for infertility and were divided into two groups:PCOS group（n=41）and normal control group（IVF-ET group due to male factors alone）（n=43）.Meanwhile,the two groups were divided into four subgroups based on body mass index（BMI）（group A1:BMI≥24 kg/m²,and TCM syndrome differentiation was phlegm-dampness syndrome;group A2:BMI<24 kg/m²,and TCM syndrome differentiation was non-phlegm-dampness syndrome;group B1:BMI≥24 kg/m²,and TCM syndrome differentiation was phlegm-dampness syndrome;group B2:BMI<24kg/m²,and TCM syndrome differentiation was non-phlegm-dampness syndrome）.Antagonist regimen and conventional controlled ovarian stimulation were used in all four groups.The outcomes of IVF-ET（including the number of oocytes retrieved,2PN,available embryos,high-quality embryos,cumulative pregnancy rate,cumulative live birth rate,etc.）were compared between PCOS group and normal control group.At the same time,wide-target metabolite data were collected using QTrap4000 triple quadrupole combined with linear ion hydrazine mass spectrometry to analyze the concentration changes of phthalates and their metabolites in follicular fluid of PCOS and normal patients to find different phthalate metabolites in PCOS with phlegm-dampness type.Results:Eighty patients were finally included in this study,including 40 PCOS patients and 40 normal controls.Analysis of clinical data showed that there were statistically significant differences in basal serum follicle stimulating hormone（FSH）,basal luteinizing hormone（LH）,and basal testosterone levels（T）between the two groups（P<0.05）.The estradiol concentration（E₂）on the day of h CG trigger was significantly higher in the PCOS group than in the normal control group（P<0.05）.The fertilization rate and cumulative live birth rate in the normal control group were higher than those in the PCOS group,and the difference was statistically significant（P<0.05）.The number of high-quality embryos and cumulative pregnancy rate in the normal control group were higher than those in the PCOS group,but the difference was not statistically significant（P>0.05）.Meanwhile,targeted metabolomics analysis showed that phthalate metabolite concentrations in follicular fluid were increased in PCOS group compared with normal control group,and the difference was statistically significant（P<0.05）.In particular,dioctyl phthalate（DNOP）,diisobutyl phthalate（DIBP）,diethyl phthalate（DEP）,dibutyl phthalate（DBP）,dibutoxyethyl phthalate（DBEP）,and dipropyl phthalate（DPr P）concentrations were significantly increased（P<0.01）.However,subgroup analysis revealed that DBP,DIBP,DPr P,DBEP,dipentyl phthalate（DPP）,dicyclohexyl phthalate（DCHP）,and butylbenzyl phthalate（BBP）concentrations were increased in the PCOS group with TCM syndrome differentiation of phlegm-dampness syndrome compared with the normal control group,and the differences were statistically significant（P<0.05）.Conclusion:The normal fertilization rate,the number of high-quality embryos,the cumulative pregnancy rate and the cumulative live birth rate of PCOS patients in the normal control group were higher than those of PCOS patients with phlegm-dampness type,indicating that PCOS has adverse effects on the IVF-ET treatment outcomes of patients.At the same time,the concentrations of phthalate metabolites（DBP,DIBP,DPr P,DBEP,DPP,DCHP,BBP）in follicular fluid of PCOS patients with phlegm-dampness type were changed,suggesting that the above substances may play an important role in the occurrence and development of PCOS with phlegm-dampness type,which is of great significance for the diagnosis and prevention of PCOS and TCM syndrome differentiation and treatment.