The Correlation Study Of ROS And Ovarian Aging

Objective: With the delay of childbearing age,the infertility rate of elderly women increases obviously,and the decline of quality of ovum is the key to affect the pregnancy outcome.Decreased ovarian function includes decreased ovarian reserve and poor ovarian response(POR).Decreased ovarian reserve function refers to the reduction of the number of original follicles,effecting the follicle growth,development,and the formation of fertilized eggs ability;POR refers to the adverse reaction of ovary to gonadotropin stimulation,resulting in low egg harvest rate and reduced pregnancy rate.However,there is no unified diagnostic standard at present.Oxidative stress is a common mechanism of aging and disease,and may be the main mechanism of ovarian aging.Reactive oxygen species(ROS)is a major indicator of oxidative stress.in this paper,based on 2016 POSEIDON standard categorizing POR crowd,to explore whether there is an obvious correlation between ROS and ovarian aging,the levels of oxygenation stress in follicles contained in different ovarian response degrees were measured.At the same time,mice were taken as the research object to explore whether hypoxia was an important factor in ovarian aging.Methods: Clinical research: recruit 100 patients from January 2018 to December 2019 in Dalian Municipal Woman and Children’s Medical Center who accepted IVF infertility clinic treatment,There were 80 patients with ovarian low reaction(age 25 to 43,attains the egg number<10,group A,B,C,D),20 patients with normal ovarian response(age 23 to 34 years old,egg number 10 or higher,group E).Stimulating ovulation with antagonist regimen,collecting relevant data,and analyzing clinical characteristics.After obtaining the informed consent of the study subjects,part of the follicular fluid was collected on the day of egg collection to measured the ROS level of follicular fluid by human ROS enzymatic immunoassay(ELISA)kit,and the correlation between ROS and IVF outcomes was analyzed.Animal experiment: 11 female SPF mice were selected as experimental group and kept in a plastic box with oxygen concentration of 10% for 4 weeks without oxygen.Another 10 female SPF mice were selected as control group and kept in a normal oxygen environment for 4 weeks.After 4 weeks,they were allowed to mate with male mice under normal oxygen conditions to record the number and quality of their offspring.8 weeks later the two groups of mice were feeding in oxygen-deficient environment for 4 weeks to stimulate ovulation and collect the ovum,then using the active oxygen fluorescence detection kit for detecting the egg ROS and fluorescence photographs taken under a microscope,with the Image-proplus 6.0 software to the fluorescence intensity Image quantitative analysis,using SPSS 22.0 statistical software for data analysis.Results: Clinical study: 1.Comparison of clinical characteristics and IVF outcomes in each group.Gn days of group D were significantly less than other 4 groups(P<0.05).Gn dosage of group B was significantly higher than that of group A,group D and the control group(P<0.05).The number of eggs harvested,the number of 2PN fertilization,the number of cleavage,the number of high-quality embryos and the rate of high-quality embryo formation in the control group were significantly higher than those in the other four groups(P<0.05).The number of eggs harvested,the number of fertilization by 2PN and the number of cleavage in group A and group B were significantly higher than that in group C and group D(P<0.05),and the number of high-quality embryos was slightly higher than that in group C and group D,with no statistical difference.The cleavage rate and quality embryo formation rate of group A and group C were slightly higher than that of group B and group D.There was no significant difference in the number of immature eggs(GV)and cleavage rate between the five groups.2.Comparison of ROS content in follicular fluid of each group.Each follicle fluid shows: the content of ROS in the control group < group A < group C < group B < group D,the control group and group A are less than the other 3 groups with statistical significance(P<0.05),but there is no statistical significance between the control group and group A.3.The correlation between follicular fluid ROS and IVF treatment outcomes.The ROS content of follicular fluid was negatively correlated with the number of harvested eggs,the number of 2PN fertilization and t cleavage(P<0.05),positively correlated with the amount of Gn,and negatively correlated with the number of harvested eggs,the rate of 2PN fertilization,the rate of cleavage,the number of high-quality embryos,and the rate of high-quality embryo formation,but without statistical significance.Animal experiments: 1.The number of litter in the experimental group was slightly higher than that in the control group(13.63±3.35vs11.90±4.56),and the average litter mass was slightly higher than that in the control group(24.30±6.37vs20.85±7.71),with no statistically significant difference.2.Detection of ROS content in mouse eggs.The ROS content in the quantified hypoxia group was slightly lower than that in the control group(15.92±0.62vs16.74±0.72),and the results were not statistically significant.Conclusion: 1.Compared with AMH and AFC,age may be more significant in evaluating ovarian function.Although ovarian reserve function may decline in younger women,the ovular quality may be higher than in older women with normal ovarian reserve function.2.The ROS content in the follicular fluid microenvironment was significantly increased in women with advanced age and worse ovarian reserve function,and was negatively correlated with the number of ovum,2PN fertilization,and cleavage.3.Short-term hypoxic feeding did not significantly reduce the fertility of mice,and there was no significant change in ROS.