抗核抗体表达对体外受精-胚胎移植的影响及机制研究

Although in vitro fertilization and embryo transfer techniques have been rapidly developed in recent years, a portion of infertile patients consistently fail to conceive despite recurrent transfers of high-quality embryos. One of the reason may be the break of the homeostasis in immune system. As a commonly seen autoantibody in women, anti-nuclear antibody targeted the important components in cell nuclear. This effect may impact the zygote division and leads to the failure of pregnant.In our previous study, we found that the presence of ANA in serum predicted an adverse IVF/ICSI outcome, primarily reflecting the reduced rates of implantation and clinical pregnancy, as well as the increased rates of early miscarriage. We hypothesize that ANA in the serum, the follicular fluid and the endometrial secretion may impact IVF-ET treatment by it adverse effect on embryo development and promote effect on basal layer cell and embryo cell damage. To verify our hypothesize, we plan to test and type ANA in serum, follicular fluid and endometrial secretion by indirect immunofluorescence assay and the liquid bead microarray to identify the impact effect of ANA by subtypes in the IVF-ET treatment. Mechanism research will be performed by culture embryos or granular cells with ANA positive serum and commercial ANA. Followed with intracellular apoptosis factors from embryos or granular cells will be tested by liquid bead microarray; immunofluorescence assay will be performed to identify the existence of ANA in the cell. Research data and clinic data will be summarized to study the impact of ANA in embryo development after IVF-ET treatment.

IVF-ET技术发展迅速，但妊娠率远未达到人们期望。生殖免疫学认为移植失败与机体免疫异常有关。抗核抗体（ANA）作为女性人群中最常见的自身抗体，可能与细胞核成分相结合影响细胞分裂，导致不孕或流产。我们既往研究提示，血清、卵泡液中ANA表达影响IVF-ET的结局，阳性组妊娠率降低，流产率增高；另外在子宫内膜分泌物中检出ANA；我们认为血清、卵泡液、子宫内膜中的ANA可能影响成功妊娠。为验证假设，我们对IVF-ET患者的血清、卵泡液、子宫内膜分泌物进行筛查，液态微球芯片法对ANA分型检测，探讨不同表达部位及不同亚型的ANA对IVF-ET的影响；将自身阳性血清、单/多克隆ANA与废弃胚胎及颗粒细胞共培养，液态微球芯片技术检测培养上清液中凋亡因子的表达，免疫荧光法检测废胚及颗粒细胞中ANA结合位点；研究ANA与胚胎靶抗原结合、对胚胎细胞凋亡的影响，以探讨其影响胚胎发育的可能机制。

近年来，生殖免疫学越来越受到重视，在影响体外受精-胚胎移植治疗结局方面的研究逐步开展。移植失败可能与机体免疫异常有关，抗核抗体（ANA）作为女性人群中最常见自身抗体，可能与细胞核成分相结合影响细胞分裂，导致不孕或流产。我们通过研究血清、卵泡液、子宫内膜中的ANA，发现血清中抗核抗体阳性影响体外受精-胚胎移植的妊娠结局，卵泡液中抗核抗体阳性可能导致更差的IVF结局。对ANA进行分型研究，发现了影响IVF结局的常见亚型。在患者子宫内膜细胞和颗粒细胞中检到ANA，提示我们ANA可以侵入生殖器官系统，从而影响胚胎的发育和着床。通过对不同人群卵泡液中凋亡因子的检测，发现ANA阳性组凋亡因子的表达与阴性组比较差异有统计学意义；细胞与单克隆ANA孵育后出现凋亡因子和生长因子表达的差异，提示ANA可能通过加速细胞凋亡或抑制生长来影响细胞的增殖和发育。我们的研究提示一方面在临床工作中应重视生殖免疫的作用，从而改善和提高IVF的治疗结局，给更多的家庭带来福音；另一方面，进一步促进基础机制研究，发现免疫治疗的切入点，更好的服务临床。