| Endometriosis(endometriosis,EMs)is a common cause of female infertility.Currently,surgery and in vitro fertilization and embryo transfer(IVF-ET)are two main methods for endometriosis patients with infertility to get pregnant.And assisted reproductive technology(ART)has increasingly become their best choice.The widely used assisted reproductive technology not only brings new hope for endometriosis patients with infertility but also provides an opportunity for the study of the special stage of reproductive process.But it has been found that,the endometriosis patients have poor embryo quality and lower pregnancy rate compared with patients with other infertile factors in the ART process.The high level of granulosa cells’ apoptosis may result in the poor quality of occyte and lower developmental potential.Objective: Through measuring the level of growth hormone(GH)in serum and follicular fluid in patients with endometriosis,detecting their expression of Bcl-2 and Bax protein,which were related to the apoptosis of granulosa cells,and detecting the effect of GH on Bcl-2 and Bax protein in patients with endometriosis,we explored the relationship between GH and endometriosis and evaluated whether GH can improve the state of granulosa cells’ apoptosis.Methods: Totally 65 patients undergoing IVF-ET were collected,including 40 patients with endometriosis(EM group)and 25 patients merely with fallopian tube obstruction(control group).All these patients were selected from the department of reproductive medicine in the second hospital of Hebei Medical University from March 2013 to September 2015.The serum,follicular fluid and luteinized granulosa cells of these patients were collected on the day of oocytes retrieval.The growth hormone in serum and follicular fluid were detected by radioactive immunosorbent assay.All the granulosa cells were detected through immunocytochemistry assay to measure the expression of Bcl-2 and Bax protein.The EMs patients’ granulosa cells were cultured in incubater,and they were devided into two groups(GH intervention group and non-intervention group)by wether a concentration of 0.15μg / ml of recombinant human growth hormone were added to the granulosa cells’ culture medium.The expression of Bcl-2 and Bax on those granulosa cells were detected through immunocytochemistry assay.Compared the clinical characteristics,laboratory characteristics,clinical outcomes,the level of growth hormone in serum and follicular fluid,and the levels of Bcl-2 and Bax in granulosa cells between EM group and control group.Results:1 Comparing the clinical characteristics between EMs and control group: there was no statistical significance on comparing the ages,years of infertility,duration of COS and the dosage of Gn between the two groups.The amount of antral follicles were 10(7)vs 13(6),and it had significantly difference(Z=-2.982,P=0.003).2 Comparing the laboratory characteristics between EMs and control group: the fertilization rate of the two groups were 80.91(26.67)% vs 84.21(30.95)%,the EM group was lower than control group,but it had no significant difference(Z=-0.726,P=0.468).The rate of available embryos were 41.42(21.00)% vs 46.42(20.51)%,The rate of good-quality embryos were 20.00(36.00)% vs 26.79(43.00)%,the EM group was lower than control group,but neither of them had significant difference(Z=-0.358,P=0.72;Z=-0.163,P=0.87).3 Comparing the clinical outcomes between EMs and control group: the biochemical pregnancy rate,clinical pregnancy rate and implantation rate of the two groups were 58.6% vs 76.0%,58.6% vs 72% and 43.6% vs 56%,the EM group was lower than control group,but neither of them had significant difference(χ2=1.825,P=0.177;χ2=1.054,P=0.305;χ2=1.602,P=0.524).4 Comparing the level of growth hormone in serum and follicular fluid of EMs and control group: the GH concentrations in serum were 1.74±0.21ng/ml vs 3.11±1.08ng/ml,the EM group was significanty lower than control group(t=-6.257,P=0.000).The GH concentrations in follicular fluid were 1.41(0.26)ng/ml vs 2.13(1.57)ng/ml,the EM group was significanty lower than control group(Z=-3.344,P=0.001).5 Comparing the levels of Bcl-2and Bax in granulosa cells of endometriosis group and the control group: the level of Bcl-2 in granulosa cells of the two groups were 0.17(0.11)vs 0.26(0.07),the EM group was significanty lower than control group(Z=-4.386,P=0.000).The level of BAX in granulosa cells of the two groups were 0.27(0.08)and 0.14(0.03),the EM group was significanty higher than control group,(Z=-5.208,P=0.000).6 Comparing the levels of Bcl-2 and Bax expression in granulosa cells of the growth hormone-treated group and non-treated group with endometriosis: the level of Bcl-2 in granulosa cells of the two groups were 0.21(0.06)vs 0.17(0.11),the growth hormone-treated group was significantly higher than the non-treated group(Z=-2.422,P=0.015).The level of Bax in granulosa cells of the two groups were 0.18±0.30 vs 0.27±0.06,the growth hormone-treated group was significantly lower than the non-treated group(t=-5.825,P=0.000).Conclusion: Endometriosis may result in reduced serum and follicular fluid growth hormone concentration and increased apoptosis particles,thus may affect the quality of oocyte and have an adverse effect on the pregnancy outcomes of IVF-ET.A certain concentration of growth hormone may improve endometriosis patients with granulosa cells’ apoptotic state. |
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