Effect Of Follicular Fluid From Patients With Emdometriosis-associated Infertility To Maturation Of Mouse Oocytes And Intervention Study Of Neiyi Recipe On The Mechanism

Endometriosis(EM)is a kind of female estrogen-dependent disease which is characterized by the ectopia implant of endometrial stromal cells and glandular epithelial cells.Incidence of EM in women of reproductive age is about 10%~15%.EM and infertility are highly relevant.Compared with other infertility patients with other factors,patients with EM-associated infertility have lower rate of normal oocytes in assisted reproductive technology(ART)and lower rate of pregnancy,which may be caused by follicular maturation disorders,impaired oocyte quality and other factors.Surrounding the oocytes,follicular fluid will inevitably affect the maturation and quality of oocytes.Studies have shown that there is a significant difference between proteomic analysis of follicular fluid from women with EM-associated infertility and that of normal female or infertility patients with other causes.In addition,researchers have studied the cytokines and proteins which are expressed abnormally and they thought that it is one of the main causes of EM-associated infertility except for pelvic anatomy disorder.Neiyi recipe is an clinical chinese traditional medicine complex prescription which is effective to treat EM and EM-associated infertility.This study aims to find new notion and treatment to EM-associated infertility by researching the cytokines and proteins which were expressed obviously abnormally,investigating the affection of EM-follicular fluid to mouse oocytes maturation and quality and exploring the effect of Neiyi Recipe drug serum on mouse oocytes maturation interfered by follicular fluid from EM patients.Objectives1.Research the effect of follicular fluid from EM-associated infertility patients on maturation and quality of mouse oocytes and the underlying mechanism.2.Investigate the cytokines and proteins which are expressed obviously abnormally in follicular fluid from EM-associated infertility patients.3.Explore the effect of Neiyi recipe drug serum on mouse oocytes maturation interfered by follicular fluid from EM-associated infertility patients.Methods1.By incorporateing follicular fluid from EM-associated infertility patients into the medium,we attempt to ascertain the interference of follicular fluid from EM-associated infertility patients to mouse oocytes maturation in vitro.The affection of follicular fluid from EM-associated infertility patients to mouse oocytes quality by fluorescent staining mature mouse oocytes spindles and observing their morphology.We understand underlying mechanism of follicular fluid from EM-associated infertility patients affecting mouse oocytes quality and maturation in vitro by fluorescent staining mouse oocytes matured in vitro which is interfered with follicular fluid from EM-associated infertility patients and detecting the average fluorescent intensity of the oocytes,which represents the level of oxidative stress.2.By protein chip detection of follicular fluid from EM-associated infertility patients,we screen out the cytokines and proteins which are expressed differently(twice or more)between group EM and group control.Then,we choose the EM-associated ones to detect and verify the different expression in follicular fluid of both groups by ELISA method with lager samples.3.Based on the participation of follicular fluid from EM-associated infertility patients to mouse oocytes in vitro maturation,we add Neiyi recipe drug serum to the in vitro maturation medium to explore the effect of Neiyi recipe drug serum on mouse oocytes maturation interfered by follicular fluid from EM-associated infertility patients.Results1.Follicular fluid from EM-associated infertility patients degrades maturing rate of mouse oocytes matured in vitro,impairs spindles and enhances the level of oxidative stress in these oocytes.2.ICAM-1 expression is significantly increased(P <0.001)in follicular fluid from EM-associated infertility patients.TECK expression is significantly decreased(P <0.001)in follicular fluid from EM-associated infertility patients.I-309,Lymphotactin,MMP-25,TSG-6 were abnormally expressed in follicular fluid from EM-associated infertility patients.3.Neiyi Recipe drug serum can improve maturing rate of mouse oocytes matured in vitro which is reduced by follicular fluid from EM-associated infertility patients.Conclusions1.Follicular fluid from EM-associated infertility patients enhances the level of oxidative stress in oocytes,interferes maturation in vitro of mouse oocytes.2.ICAM-1 is significantly overexpressed in follicular fluid from EM-associated infertility patients and it possibly results the disorder in follicular fluid microenvironment with other cytokines and proteins expressed abnormally.3.Neiyi recipe drug serum could improve the interfered maturation of mouse oocytes impaired by follicular fluid from EM-associated infertility patients.