| Objective10–15% women in reproductive age are affected by endometriosis(EM), at least 50% women combined with infertility. EM patients have poorer clinical outcomes of assisted reproductive technology(ART) but specific theoretical basis is unclear. Because of the lesions mainly located in the ovary which is the place of development and maturation of oocytes, ovarian Endometriosis(OEM) is likely to have great impact on oocyte maturation and fertilization and in our studies certain culture conditions are analyzed. The changes of oocyte morphology maybe associated with the maturity and development potential of oocytes. We analyze the incidence rate changes of the ooctyes with normal morphology, thick pellucid zone, large perivitelline space, coarse granule, broken polar body and clinical outcomes changes of In vitro fertilization and embryo transfer(IVF-ET), in order to provide experimental basis for predicting clinical outcome through oocyte morphology. The impact of EM follicular fluid microenvironment changes on the quality of oocyte and granulosa cells becomes a hot spot in ART field. We analyze the effects of changes of inflammatory cytokines IL-6, TNF-α, IL-1 in the follicular fluid of OEM patients on oocyte maturation, differentiation, granular cell proliferation, providing experimental basis for improving OEM clinical outcome. MethodsThe data of 152 OEM patients in Tangshan Maternal and Child Health Hospital IVF Center were collected retrospectively from June, 2013 to January, 2015. 168 cases tubal factor infertility were used as control group. Basic clinical data including age, infertility years, body mass index, antral follicle numbers, FSH, LH, E2 of two groups were compared. Clinical outcomes including oocyte retrieval rate, MII oocyte rate, fertilization rate, R-ICSI rate, good quality embryo rate, biochemical pregnancy rate, clinical pregnancy rate, abortion rate were compared. The incidence rates of normal morphology, thick pellucid zone, large perivitelline space, coarse granule, broken polar body were compared between two groups. Fertilization rate, R-ICSI rate, good quality embryo rate, biochemical pregnancy rate, clinical pregnancy rate of five morphology oocytes in OME group were compared reciprocally. The changes of IL-6, TNF-α, IL-1 in follicular fluid were detected by ELISIA. Mouse immature oocytes in different stages were cultured in two different follicular fluids. The mature rates at 6h,12 h,24h,48 h was compared. Abnormal morphology rate,fertilization rate and good quality embryo rates of oocytes matured in vitro and in vivo were compared. Culture conditions of cumulus granular cells were observed and apoptosis rate of cumulus granular cells were detected by TUNEL and the changes of single follicle estrogen level were detected by chemiluminescence. The correlations between IL-6, TNF-α, IL-1 level, apoptosis rate of cumulus granulosa cells, single follicle E2 level, MII oocyte rate and clinical pregnancy rate were analyzed. Results1.There was no significant difference between OEM and tubal factor infertility group on basic clinical data(P>0.05). Oocyte retrieval rate, MII oocyte rate, fertilization rate, good quality embryo rate, biochemical pregnancy rate, clinical pregnancy rate of OEM group were significantly lower than those of tubal factor infertility group(P<0.05). R-ICSI rate and abortion rate of OEM group were significantly higher than that of tubal factor infertility group(P<0.05)2.The oocytes with normal morphology in OEM group were significantly decreased compared to tubal factor infertility group, while the oocytes with coarse granule, large perivitelline space, thick pellucid zone in OEM group were significantly increased compared to tubal factor infertility group( P < 0.05). Compared to tubal factor infertility group, the oocytes with broken polar body were not increased significantly(P>0.05). Good quality embryo rate, biochemical pregnancy rate, clinical pregnancy rate in normal morphology group were significantly increased compared to coarse granule, large perivitelline space, broken polar body group(P<0.05). The fertilization rate with normal morphology oocytes was significantly increased while R-ICSI rate was significantly decreased compared to thick pellucid zone group(P<0.05). All data of coarse granule, large perivitelline space, broken polar body group have no significant difference compared to each other(P>0.05)3.The levels of IL- 6, TNF-α, IL- 1 in OEM group were significantly higher than tubal factor infertility group(P < 0.05). The mature rates of GV, MI, AM reached to the peak at 24h、12h、6h respectively. The mature rate in OEM group was significantly decreased compared to tubal factor infertility group. Fertilization rate and abnormal morphology rates of oocytes matured in vitro were not significantly decreased(P>0.05). But good quality embryo rate was significantly decreased when compared to oocytes in vivo(P<0.05).4.The culture conditionn of cumulus granular cells in OEM group was poor. Cumulus granular cell apoptosis rate of OEM group was significantly increased while single follicle estrogen level was significantly decreased when compared to tubal factor infertility group(P<0.05). The levels of IL- 6, TNF-α, IL-1 were positively correlated with cumulus granular cell apoptosis rate; Cumulus granular cell apoptosis rate was negatively correlated with single follicle E2 level; Single follicle E2 level was positively correlated with MII oocyte rate and MII oocyte rate was positively correlated with clinical pregnancy rate(P < 0.001). ConclusionsThe clinical outcomes in OEM patients were poor and abnormal morphology oocytes were increased. The oocytes with different abnormal morphology had different developmental potential. The levels of IL-6、TNF-α、IL-1 in follicular fluid significantly increased and resulted in flammatory microenvironment, which might affect oocyte maturation, differentiation and development. Different immature eggs needed different culture time in vitro. Cumulus granular cell apoptosis increased in inflammatory follicular fluid, declined the synthesis of estrogen and then affected oocyte maturation and differentiation. We could improve oocyte quality and its inflammatory microenvironment to improving OEM IVF-ET results... |
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