| Endometriosis(EMs) is one of the most common cause of female infertility.Assisted reproductive technology is widely used, which not only brings the hope of birth for the infertility patients, but also provides an opportunity for the study of the particular stage of the reproductive process. But compared with patients of other infertile factors, the patients with endometriosis have the poor pregnancy outcome of undergoing in vitro fertilization and embryo transfer(IVF-ET). The reasons of adverse outcome by EMs are complicated. In recent years, it has been found that the oxidative stress(OS) may be related to the occurrence of endometriosis. The maintain of cellular functions needs oxygen, but its metabolites, such as reactive oxygen species(ROS), can change the cell function and(or) endanger cell survival. The body has the ability of getting rid of excess ROS and making ROS to maintain a certain level, which is called the total antioxidant capacity(TAC). In a healthy body, reactive oxygen species and antioxidants are in balance. However, when this balance is destroyed, excessive reactive oxygen species exist, then the oxidative stress occur. Excessive ROS can lead to nuclear DNA and mitochondrial DNA’ mutation or deletion, cell membrane’ lipid peroxidation which cause the change of membrane fluidity, and protein’ oxidative damage which cause important enzyme inactivation. The oxidative stress may be one of the infertile causes associated with endometriosis, but the relationship between oxidative stress and endometriosis remains controversial.Objective: Through measuring the level of ROS and TAC in the serum, follicular fluid and granulosa cell of the patients with endometriosis, and calculating the ratio of ROS/TAC, at the same time, combining with the various assisted reproductive technology index, we evaluated whether oxidative stress affected the pregnancy outcome in patients with infertility associated with endometriosis, as well as investigated the relationship between oxidative stress and endometriosis.Methods: 30 patients with endometriosis(group A) and 30 patients with fallopian tube obstruction(group B) undergoing IVF-ET were collected. All these patients were selected from the department of reproductive medicine in the second hospital of Hebei Medical University from November 2013 to May 2014. The serum, follicular fluid and luteinized granulosa cells of these patients were collected. Then measure the level of ROS and TAC in serum, follicular fluid and the granulosa cells by enzyme-linked immunosorbent assay(ELISA), and calculate the ratio of ROS/TAC. Compare two groups in clinical characteristics, laboratory indexes, clinical outcomes, the level of ROS, the level of TAC and the ratio of ROS/TAC. Analyze the correlation between the level of ROS, level of TAC, ratio of ROS/TAC and the age, years of infertility, basal hormone levels, duration of controlled ovarian stimulation(COS), dosage of gonadotropin(Gn), number of eggs, number(or rate) of fertilization, number(or rate) of diploid zygotes, number(or rate) of embryo cleavage, number(or rate) of available embryos, number(or rate) of good-quality embryos.Results:1 Comparing the clinical characteristics between group A and group B: the dosage of Gn about group A and B was 2768.13±640.54 IU vs 2248.13±634.59 IU, and it had the statistical significance(t=-2.579, P<0.05). However, there was no statistical significance on comparing the ages, years of infertility, basal hormone levels and duration of COS.2 Comparing the laboratory indexes, clinical outcomes between group A and group B: the number of good-quality embryos about group A and B was 1.00(4.00) vs 4.00(4.00), as well as the rate of available embryos and rate of good-quality embryos were 41.20% vs 51.80%, 22.40% vs 33.21%, and all the difference had statistical significance(Z=-1.980, P<0.05; χ2=5.928, P<0.05; χ2=7.571, P<0.05). The clinical pregnancy rate and implantation rate of group A and B were 25.00% vs 34.40%, 41.20% vs 50.00%, but it had no statistical significance.3 Comparing the level of ROS, level of TAC, ratio of ROS/TAC in serum between group A and group B: the level of ROS and the ratio of ROS/TAC in group A and B respectively were 0.97(0.56)ng/ml vs 1.09(0.47)ng/ml and 0.73(0.51) vs 0.97(0.56), but it had no statistical significance. The level of TAC in group A and B was 1.52±0.41 ng/ml vs 1.26±0.27 ng/ml, and it had statistical significance(t=-2.393, P<0.05).4 Comparing the level of ROS, level of TAC, ratio of ROS/TAC in follicular fluid between group A and group B: the level of ROS, level of TAC and ratio of ROS/TAC in group A and B respectively were 1.08(0.67) ng/ml vs 0.81(0.46)ng/ml, 1.02±0.29ng/ml vs 1.44±0.30ng/ml, 1.20(0.88) vs 0.61(0.39), and there was statistical significance(Z=-2.407, P<0.05; t=4.460, P<0.05; Z=-3.733, P<0.05).5 Comparing the level of ROS, level of TAC, ratio of ROS/TAC in granulosa cells between group A and group B: the level of ROS, the level of TAC and the ratio of ROS/TAC in group A and B respectively were 0.92(1.03)×10-6ng/cell vs 1.07(0.32)×10-6ng/cell, 1.43±0.76×10-6ng/cell vs 1.19±0.57×10-6ng/cell, 0.74(0.80) vs 1.05(0.45), but there was no statistical significance.6 The correlation between the level of ROS, level of TAC, ratio of ROS/TAC and the clinical characteristics: in follicular fluid, the level of ROS had a positive correlation with the ages of patients, duration of the infertility and dosage of Gn(rs=0.524, P<0.05; rs=0.298, P<0.05; rs=0.331, P<0.05). The level of TAC had a negative correlation with the dosage of Gn(rs=-0.336, P<0.05). The ratio of ROS/TAC was positively correlated with the ages of patients and dosage of Gn(rs=0.438, P<0.05; rs=0.360, P<0.05).7 The correlation between the level of ROS, level of TAC, ratio of ROS/TAC and the laboratory indexes: in follicular fluid, the level of ROS was negatively correlated with the fertilization rate(rs=-0.240, P<0.05), and the ratio of ROS/TAC had a negative correlation with the fertilization rate, number of available embryos and number of good-quality embryos(rs=-0.293, P<0.05; rs=-0.281, P<0.05; rs=-0.302, P<0.05). In granulosa cells, the level of ROS had a negative correlation with the fertilization rate and rate of diploid zygotes(rs=-0.409, P<0.05; rs=-0.440, P<0.05); the level of TAC was negatively correlated with the fertilization rate, rate of diploid zygotes and rate of good-quality embryos(rs=-0.320, P<0.05; rs=-0.379, P<0.05; rs=-0.313, P<0.05).8 In follicular fluid, the level of ROS, level of TAC and ratio of ROS/TAC in pregnant group and non-pregnant group respectively were 0.99(0.38)ng/ml vs 1.06(0.68)ng/ml, 1.26±0.38ng/ml vs 1.24±0.38ng/ml,0.63(0.93) vs 1.03(0.59), but there was no statistical significance.Conclusion:Endometriosis may have an adverse effect on the pregnancy outcomes of IVF-ET,reduce the ovarian reactivity to gonadotropin,and affect the quality of oocyte.Oxidative stress increases in the follicular fluid of patients with endometriosis,which may affect the development potential of oocyte.The oxidative stress markers in follicular fluid may be the predicted index for pregnancy outcome.PCR polymerase chain reaction聚合酶链反应NO nitric oxide一氧化氮MMP mitochodria membrane potential线'体膜电位... |
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