The Effect Of Estrogen And Related Inflammatory Factors On The Development Of Follicle

Estrogen plays an important role in the development of follicle. Our research aimed to elucidate the association and interaction among estrogen and inflammatory factors involved in the development of follicle, and evaluate the affects of these factors to the outcome of assisted reproductive technology and the proliferation of the granulosa cells in vitro.Section I:Endocrine and inflammatory factors and endometriosis-associated infertility in clinically assisted reproductionObjective:Assisted reproduction techniques are commonly used for endometriosis-associated infertility, but the outcome of ART pregnancy is effected by endometriosis. Our research aimed to evaluate the effects of endometriosis on folliculogenesis and pregnancy and to assess the involvement of inflammatory factors (IL1β, PGE2, PGF2a, and TGFβ2) within the follicular fluids.Methods:This is a case-control study involving29patients with endometriosis and36controls undergoing ART by standard ovarian stimulation protocols at Renji Hospital. A total of65follicular fluid aspirates were collected. The concentrations of IL1β, PGE2, PGF2a, and TGFβ2within the follicular fluid and serum E2concentration are measured, the mRNA expression of IL1β, Ptgs2, aromatase and PPARy in the granulosa cells are determined and ART outcomes are checked up in the followed-up.Results:The number of oocyte retrieval, the rate of MII occytes, the rate of cleavage zygote, the rate of effective embryo and pregnancy rates of patients with endometriosis were significantly lower than those of control patients. In patients with endometriosis, the serum E2concentration measured2days before oocyte retrieval was positively correlated with the number of oocytes retrieved and was lower than in control patients. Aromatase levels in the granulosa cells of the endometriosis group were lower, and the concentrations of PGE2in the follicular fluid were higher than in controls. Furthermore, progesterone levels were significantly lower in the follicular fluid of the endometriosis group. The concentrations of PGE2, PGF2α, TGFβ2and IL1β were significantly correlated with each other; PGE2was positively correlated with endometriosis; and endometriosis negatively correlated with endocrine levels and pregnancy rates.Conclusions:These results suggest that ART outcomes, related to the serum E2concentration measured2days before, impaired by endometriosis. Furthermore, endocrine and inflammatory factors within the follicular fluid mediate the effects of endometriosis by reducing the number of oocytes retrieved and decreasing pregnancy rates in clinically assisted reproduction. Section Ⅱ:The effects of endocrine and inflammatory factors in the follicular fluid from PCOS patients undergoing GnRH-a/FSH program on the outcome of ARTObjective:Our research aimed to evaluate the effects of PCOS on folliculogenesis and pregnancy, and to assess the involved contribution of inflammatory factors in the follicular fluids, such as IL1β, PGE2, PGF2α, and TGFβ2.Methods:This is a case-control study involving17patients with PCOS and16controls undergoing ART by GnRH-a/FSH program at Renji Hospital. The concentrations of IL1β, PGE2, PGF2α, and TGFβ2in the follicular fluid, and serum E2concentration, are measured by ELISA; the mRNA expression of IL1β, Ptgs2, aromatase and PPARy in the granulosa cells are determined by realtime PCR; and ART outcome were checked up and recorded in the followed-up.Results:The oocyte retrieval and pregnancy rates of patients with PCOS were similar to those of control patients, however, the concentrations of IL-1β, PGE2and TGF-β2in the follicular fluid were higher than that in the controls.Conclusions:These results suggest that the elevated inflammatory factors in the follicular fluid may reflect the pathophysiologic mechanism of PCOS, but does not impair the ART outcome. So, the GnRH-a/FSH program could reverse the endocrine disorder in PCOS, and improve pregnancy rates in clinically assisted reproduction.Section Ⅲ:The effect of estrogen on the secretion of inflammatory factors in the murine granulosa cells cultured in vitroObjective:Observe the proliferation of mouse granulosa cells after adding estradiol in vitro, measure the expression of IL-1β and TGF-β2which involved in the development of granulosa cells.Methods:4weeks immature mouse granulosa cells were collected and cultured. After these cells adhered to the bottom of culture plate, in the study group added1ul estradiol (100ng/nl) which dissolved by DMSO in the experimental group, the same volume of DMSO was added in control. The expression of IL-1β、TGF-β2mRNA are measured after48hours.Results:The cultured granulosa cells express varieties of steroid receptors, such as follicle stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), estrogen receptor alpha (ERa) and estrogen receptor beta (ERP). The expression of FSHR descresed after24hours adhension. After adding estradiol48h, the number of cell proliferation in the experimental group compared showed no significant difference compared with control group. And the expression of IL-1β、TGF-β2mRNA decreased in the study groups.Conclusions:The expression of FSHR mRNA of the cultured granulosa cells decreased after losing simulation by FSH. And estradiol on the level of100ng/ml have no significant promotive effect on granulosa cells’proliferation but reduced the expression of IL-1β、TGF-β2in the granulosa cells.