IVF Outcome And Follicular Fluid Metabolomics Study On Acupoint Sticking Combined With Traditional Chinese Medicine Intervention In EMs Of Kidney Deficiency And Blood Stasis Type

Objective:To evaluate the effect of acupoint sticking combined with traditional Chinese medicine on the clinical curative effect of kidney deficiency and blood stasis endometriosis(endometriosis,EMs)patients undergoing in vitro fertilization-embryo transfer(in vitrofertilizationembryo transfer,IVF-ET).The possible mechanism of action was explored from the perspective of non-targeted metabolomics of follicular fluid.Methods:The 84 patients who underwent IVF-ET at the Reproductive and Genetic Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine due to infertility were chosen for the clinical research using a prospective,double-blind,randomized,and controlled research methodology.According to SPSS25.0,56 cases of EMs infertility patients with kidney disease and blood stasis were randomly assigned to treatment and placebo groups.Of these,28 cases received acupoint application along with traditional Chinese medicine treatment,28 cases received acupoint application along with Chinese medicine placebo treatment,and the remaining 28 cases served as the blank control group who received no treatment.All three groups received a Gn RH antagonist fixed regimen for ovulation induction,acupoint sticking in conjunction with traditional Chinese medicine,and placebo treatment.Acupoint sticking began seven days prior to the ovulation induction cycle.The three groups’ IVF treatment outcomes(number of retrieved eggs,total number of normally fertilized eggs,normal fertilization rate,total number of high-quality embryos,and embryo implantation rate)as well as the differences in TCM syndrome scores between the treatment group and the placebo group were compared.On the day of egg extraction,the three patient groups’ follicular fluid was subjected to non-targeted metabolomics analysis using a liquid chromatography-mass spectrometry analysis system(UPLC-MS/MS)in the experimental investigation.Screening out significantly different metabolites using metabolite hierarchical cluster analysis,multivariate statistical analysis,standardized treatment,Principal Component Analysis,Partial Least Squares-Discriminant Analysis,and other analysis and development methods.Analyzing related metabolic pathways using KEGG pathways.Results:1.A total of 81 patients,including 27 in the treatment group,26 in the placebo group,and 28 in the blank control group,successfully completed the clinical study.2.The fundamental information for the three patient groups did not significantly differ,with the exception of parity and the type of infertility(P<0.05).The treatment group’s improvement in kidney insufficiency and blood stasis syndrome was more pronounced than it was in the placebo group(P<0.05).3.The time of Gn use,total amount of Gn use,number of retrieved eggs,total number of normally fertilized eggs,number of transferable embryos,number of high-quality embryos,normal fertilization rate,and number of non-transferable embryos were all significantly different among the three groups(all P<0.05).4.Nontargeted metabonomics analysis results revealed that the follicular fluid of the treatment group was closer to the blank control group,suggesting that acupoint application in conjunction with traditional Chinese medicine has a certain therapeutic effect.The follicular fluid of normal women and EMs infertile patients with kidney failure and blood stasis type contained 26 distinct metabolites,including lipids,organic acids,and organic heterocycles.5.The seven metabolites of 4-en-21-al,Testosterone acetate,Glycodeoxycholic acid,and Biliverdin all changed from up-regulated to down-regulated after acupoint application combined with traditional Chinese medicine intervention.These metabolites are primarily involved in ether lipid metabolism,the biosynthesis of phenylalanine,tyrosine,and tryptophan,and Steroid hormone biosyn.Conclusion:1.The clinical symptoms and treatment outcomes of EMs patients with kidney insufficiency and blood stasis can be effectively improved by acupuncture point sticking in conjunction with traditional Chinese medicine.2.Patients with kidney disease and blood stasis EMs and healthy women differ in the follicular fluid metabolites of 9-cis,11-transOctadecadenoate,(6Z,9Z,12Z)-Octadecatrienoic acid,hopane-29-acetate,erucamide,and other compounds.L-Threonate,Glycerophosphocholine,and 4-(3-Hydroxy-2-naphthyl)-2-oxobut-3-enoic Acid 8-Amino-7-oxononanoate,Acid glycocholic,1-Staroyl-sn-glycerol,17-hydroxyprogesterone 3-19 phosphocholine metabolites DL-O-tyrosine,L-Arginine,L-Tyrosine,Testosterone Acetate,Glycodeoxycholic Acid,Biliverdin,Dammara-20,24-diene,and Oleamide are all components of 20alpha-Hydroxy-4-pregnen-3-one.It is believed to be a biomarker of EMs of the kidney deficit and blood stasis type since it is closely associated to EMs.3.The biosynthesis of phenylalanine,tyrosine,and tryptophan,as well as the biosynthesis of steroid hormones,may be linked to the mechanism of acupoint application in combination with traditional Chinese medicine intervention.