Relationship Between Nitric Oxide (NO) in Follicular Fluid and Sperm Fertilization Ability
试验速览
- 阶段
- 不适用
- 发起方
- 入组人数
- 82
- 试验地点
- 2
- 主要终点
- Nitric Oxide levels in follicular fluid
研究概览
简要总结
Several studies indicate that Nitric Oxide (NO) plays an important role in the physiology of the reproductive system in mammals. It has been shown that NO affects sperm motility, it regulates the tyrosine phosphorylation of different sperm proteins, it enhances the sperm binding ability to the zona pellucida and it modulates the acrosome reaction.
The enzyme responsible for NO synthesis, the Nitric Oxide Synthase (NOS), has also been identified in the oocytes, cumulus and corona cells, as well as in the oviduct. For these reasons, the NOS presence at the fertilization site could be a key element to determine the success of this process. Therefore, carrying out in vitro studies to better understand NO's role in the fertilization process, especially in human sperm capacitation, could improve the outcome of the Assisted Reproductive Techniques (ARTs) due to an improvement both in the diagnosis of infertility and in the prognosis of treatment success.
This study is carried out in collaboration with the Animal Physiology Department from the Veterinary Faculty (University of Murcia, Spain) and it is funded by the European Commission under the Horizon 2020 Programme.
详细描述
Main Objective:
Determine changes in the NOS expression and protein nitrosylation in fertile and infertile patients, when adding L-Arginine, the substrate for NO synthesis, to the sperm capacitation medium in the presence/absence of human follicular fluid (hFF).
Secondary Objectives:
- Investigate if the effects of hFF on sperm capacitation are associated to NO levels in the fluid.
- Identify the correlation between NO levels in hFF and the clinical outcomes from ARTs.
METHODOLOGY
研究设计
- 研究类型
- Interventional
- 分配方式
- Non Randomized
- 干预模型
- Parallel
- 主要目的
- Diagnostic
- 盲法
- None
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •This study will include fertile males such as the semen donors attending the clinic IVI Murcia and a group of subfertile males composed of patients, who attend the clinic to undergo an in vitro fertilization treatment and who possess one altered spermiogram parameter (according to the WHO 2010 guidelines). These participants will be recruited to examine the modifications in the NOS expression as well as the protein nitrosylation in fertile and subfertile men.
- •This study will also include couples who undergo an IVF or ICSI treatment, in order to investigate if the effects of human follicular fluid on sperm capacitation are associated to NO levels in the fluid and to identify the correlation between NO levels in the follicular fluid and the clinical outcomes from ARTs.
排除标准
- •From this study will be excluded couples who undergo preimplantation genetic diagnosis, were diagnosed with repetitive abortion (three or more consecutive abortions before week 20) or implantation failure (lack of pregnancy after three embryo transfers with good quality embryos in women under 36 years of age or after two embryo transfers in women over 36 years of age), as well as women over 40 years of age.
研究组 & 干预措施
Control (fertile semen donors)
Sperm from fertile men (donors who attend the IVI Murcia clinic) will be included in this arm.
干预措施: Control (fertile semen donors) (Other)
Patients (subfertile men)
Sperm from patients (subfertile men, i.e. men who possess one altered spermiogram parameter, according to the WHO 2010 guidelines) will be included in this arm.
干预措施: Patients (subfertile men) (Other)
结局指标
主要结局
Nitric Oxide levels in follicular fluid
时间窗: Nitrit Oxide levels will be measured within 30 min after the follicular fluid sample is available.
Upon collection and after oocyte removal, the follicular fluid samples will be centrifuged to remove cellular debris. An aliquot will be further centrifuged in tubes with 10 kDa filters to remove proteins. This alliquot will then be used to determine NO concentration with the help of a commercially available measurement system. NO is rapidly oxidized into two stable ions, namely nitrite and nitrate, which can be assayed by using ions selective electrodes. A software will then be used to calculate the initial concentration of Nitric Oxide (micromolar).
次要结局
未报告次要终点