| Endometriosis(EMs)often occurs in women of reproductive age and seriously affects their fertility.Assisted reproductive technology(ART)is an important means to treat EMs infertility.Studies have shown that in vitro fertilization-embryo transfer(IVF-ET)treatment of EMs infertility can achieve a high cumulative pregnancy rate,but still lower than other causes of infertility patients,the reason may be that the rate of granulosa cell apoptosis in EMs patients increased,resulting in a decline in oocyte quality,affecting embryonic development potential,and adverse effects on pregnancy outcome.With the continuous application of growth hormone(GH)in ART,whether GH can improve the assisted pregnancy outcome in EMs patients has become the goal of the study.Objective: By detecting the apoptosis of granulosa cells in patients with endometriosis after growth hormone pretreatment and combining with their in vitro fertilization outcomes,we explored the possible mechanism of growth hormone to improve oocyte quality in patients with endometriosis.Methods:1.Retrospective analysis of 120 cycles of EMs infertile patients who received IVF-ET assisted pregnancy treatment in the reproductive medicine department of the second hospital of Hebei Medical University from January 2017 to December 2019,according to the use of GH intervention during ovarian stimulation(COS),divided into GH intervention group(A group)and non-intervention group(B group)each 60 cycles.Compare the general data,clinical indexes,laboratory indexes and embryo transfer outcomes of the two groups.2.44 cycles were the patients with EMs from September to December 2019 and divided into A1 and B1 groups,22 cycles each,according to whether they used GH or not.The level of apoptosis and mitochondrial membrane potential(MMP)in granulosa cells were detected by flow cytometry,and the level of insulin-like growth factor-1(IGF-1)in serum and follicular fluid were determined by enzyme-linked immunosorbent assay,so we can compare the level of the apoptosis and MMP of granulosa cells and the level of IGF-1 in serum and follicular fluid of A1 and B1 groups.3.The granulosa cells cultured in vitro were divided into four groups according to the GH of use: growth hormone action 24 h(L-24 h)group,growth hormone action 48(L-48 h)group and two time-matched control groups,namely 24 control(C-24h)group,48 control(C-48h)group.Apoptosis level and MMP level of granulosa cells were detected by flow cytometry to compare the apoptosis level and MMP level of granulosa cells of each group.Results: 1.Comparison of general conditions between A and B groups: there was no significant difference in age,infertility years,body mass index,basal hormone level,number of antral follicles between two groups.Comparison of clinical indicators between A and B groups: there was no significant difference in COS plan,dosage and days of gonadotropin(Gn),hormone level and endometrial thickness the day when patients injected HCG between the two groups.Comparison of laboratory indicators between A and B groups: the number of fertilized embryos,the number of transplantable embryos,the number of high-quality embryos,the rate of high-quality embryos of two groups were 9(6)vs 7(5)?3(3)vs 2(2)?2(3)vs 1(2)?33.85% vs 24.36%,A group was significant higher than B group(Z=-2.013,P=0.044;Z=-2.556,P=0.011;Z=-3.034,P=0.002;?2=8.500,P=0.004).The number of oocytes,the fertilization rate,2PN fertilization rate,2PN cleavage rate and transplantable embryo rate of two groups were 10.5(8)vs 9.53±5.13?82.91% vs 81.47%,64.41% vs 61.89%,99.12% vs 98.59%,49.12% vs 45.27%,A group was higher than B group,but the difference was not statistically significant.Comparison of outcomes of embryo transfer between A and B groups: the planting rate and clinical pregnancy rate of two groups were 37.18% vs 31.76%,48.78% vs 47.73%,A group was higher than B group,but the difference was not statistically significant.2.The IGF-1 level of serum and follicular fluid of A1 group and B1 group were 133.52±18.35ng/ml vs 120.18±21.12ng/ml?121.19±23.49ng/ml vs 105.43±23.95ng/ml,A1 group was significantly higher than B1 group(t=2.237,P= 0.031;t=2.204,P=0.033).The apoptosis rate of granulosa cells of A1 group and B1 group were 24.31±9.80% vs 28.36±6.51%,A1 group was lower than B1 group,but the difference was not statistically significant.The level of MMP of A1 group and B1 group were 33.12±8.63% vs 39.68±10.14%,A1 group was significantly higher than B1 group(t=-2.310,P=0.026).3.Comparison of the apoptosis rate and level of MMP of granulosa cells cultured in vitro: the apoptosis rate and mitochondrial depolarization ratio of granulosa cells in L-24 h group and L-48 h group were respectively lower than those in C-24 h group and C-48 h group,but the difference was not statistically significant.The apoptosis rate and mitochondrial depolarization ratio of granulosa cells in L-24 h group and C-24 h group were respectively lower than those in group L-48 h and C-48 h group,but the difference was not statistically significant.Conclusions:The application of growth hormone in control ovarian stimulation of in vitro fertilization-embryo transfer in patients with endometriosis can improve the high-quality embryo rate and improve the pregnancy outcome of in vitro fertilization-embryo transfer.The mechanism may be related to GH inhibition of ovarian granulosa cell apoptosis and further improvement of oocyte quality,and GH may act on ovarian granulosa cells by increasing IGF-1 level. |
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