To Investigate The Correlation Between PFAS Metabolites In Follicular Fluid And The Pathogenesis Of POR Based On The Theory Of "Liver And Kidney Homology" And The Intervention Of Traditional Chinese Medicine

Objective: High-throughput target metabolomics was used to qualitatively and quantitatively analyze per-and polyfluoroalkyl substances(PFAS)metabolites in follicular fluid of women with poor ovarian response(POR)of kidney deficiency and liver depression and blank control group to explore PFAS markers in follicular fluid of POR pathogenesis of kidney deficiency and liver depression;at the same time,the syndrome essence of POR kidney deficiency and liver depression was verified by analyzing the changes of PFAS metabolites in follicular fluid of POR patients before and after the intervention of Bushen Shugan herbs,and finally the pathogenesis of POR of kidney deficiency and liver depression was explored from the perspective of environmental endocrine disruptors.Methods: In the PFAS metabolite analysis study of follicular fluid in POR with kidney deficiency and liver depression,the clinical data of 62 patients who visited the Reproductive and Genetic Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from January 2019 to September 2021 for in vitro fertilization-embryo transfer(IVF-ET)assisted pregnancy were collected,of which 31 patients with POR with kidney deficiency and liver depression were used as the non-traditional Chinese medicine intervention group(POR-NON group);another 31 women who underwent IVF-ET assisted pregnancy due to male factors alone were selected as the blank control group(POR-CON group).Patients in both groups underwent controlled ovarian hyperstimulation using an antagonist protocol,and follicular fluid was retained on the day of oocyte retrieval.EXIONLC System(SCIEX)6500 QTRAP+ triple quadrupole mass spectrometer was used to detect PFAS metabolites in samples.A total of 59 cases of follicular fluid,30 cases of POR-NON group and 29 cases of POR-CON group were detected.Univariate statistical analysis and multivariate statistical analysis were used to visualize and analyze the differential metabolites.In the study of PFAS metabolites in follicular fluid after POR with kidney deficiency and liver depression treated with traditional Chinese medicine,31 patients with POR with kidney deficiency and liver depression who underwent IVF-ET assisted pregnancy at the Reproductive and Genetic Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from October 2021 to October 2022 were included as the traditional Chinese medicine intervention group(POR-TCM group)and underwent controlled ovarian hyperstimulation with antagonist protocol,and the POR-TCM group began to use Qizi Yusi Decoction for conditioning from the 5th day of the menstrual cycle before ovarian hyperstimulation until the trigger day to observe the changes of TCM syndrome scores,clinical evaluation indicators and PFAS metabolite levels in follicular fluid in the POR-TCM group and POR-NON group,and a total of 29 samples were submitted as before the experimental method.Results: The contents of PFBA in follicular fluid of POR patients with kidney deficiency and liver depression were significantly higher than those in the blank control group(P<0.01).The contents of PDA,PFOS,PFNA,PFOA and PAUn A in follicular fluid of POR patients with kidney deficiency and liver depression showed an upward trend,but the difference was not statistically significant(P>0.05).The content of PFBA in follicular fluid of POR patients with kidney deficiency and liver depression after Bushen Shugan herbs intervention was significantly decreased,compared with POR patients without traditional Chinese medicine intervention,the difference was statistically significant(P < 0.05),the content of PDA,PFOS,PFNA,PFOA,PAUn A in follicular fluid of POR patients after traditional Chinese medicine intervention showed a downward trend,but the difference was not statistically significant(P > 0.05);the total score of kidney deficiency and liver depression syndrome in POR patients after Bushen Shugan herbs intervention was significantly lower than that before treatment,the difference was statistically significant(P<0.001),and significantly lower than POR-NON group,the difference was statistically significant(P<0.05);the number of oocytes retrieved and the number of 2PN fertilization in POR-TCM group was significantly increased compared with POR-NON group,the difference was statistically significant(P<0.05);the number of non-transferable embryo cycles in POR-TCM group was significantly lower than POR-NON group(P<0.05);There were no statistically significant differences in clinical parameters such as transferable embryo rate(75.64% vs.73.02%),high-quality embryo rate(24.36% vs.22.22%)and cumulative clinical pregnancy rate(35.48% vs.32.26%)between the two groups(P>0.05).Conclusion: The level of PFAS metabolites in follicular fluid of POR patients with kidney deficiency and liver depression was significantly up-regulated compared with the blank control group.After the intervention of Bushen Shugan herbs,the TCM syndrome of POR patients was significantly improved and the number of IVF oocytes retrieved and fertilization were significantly increased,and the level of PFAS metabolites in follicular fluid was significantly down-regulated.PFAS exposure had a certain correlation with the incidence of POR with kidney deficiency and liver depression.