lncRNA-MALAT1对子宫内膜异位症不孕患者卵巢功能的调控作用及机制研究

Endometriosis (EMT) is a common chronic disease in women of reproductive age, characterized by progressive dysmenorrhea, menstrual abnormalities and infertility. In vitro fertilization and embryo transplantation is an effective method for assisted reproduction of EMT, however, decreased ovarian reserve function resulted from EMT affect the results of controlled ovarian hyperstimulation. Our previous study showed that the expression of lncRNA MALAT1 was down-regulated in mural granulose cells from women with EMT, compared with those from women with normal ovarian reserve. The expression of MALAT1 was found to have a positive correlation with antral follicle count, progestational hormone, oocytes and available embryos. In vitro studies suggested that MALAT1 may affect the expressions of the cell cycle protein inhibiting factors, p21 and p53, through ERK/MAPK signaling pathway and therefore affect cell proliferation of granulosa cells. In this project, granulose cells of follicular fluid from EMT infertility patients are collected and KGN cell line and mouse model will be used to explore the regulatory roles of lncRNA MALAT1 involved in ovarian function and the mechanism of infertility associated with endometriosis. We will also construct the MALAT1-miRNA-mRNA regulatory network and it would be helpful for the protection of women's reproductive function.

子宫内膜异位症（EMT）是育龄妇女常见的慢性疾病，表现为进行性痛经、月经异常、不孕等。体外受精-胚胎移植是EMT的有效助孕方法，然而EMT造成卵巢储备功能下降影响了超促排卵的效果。我们前期研究发现：在EMT不孕患者卵泡液颗粒细胞中lncRNA MALAT1表达水平明显下调，且其表达水平与患者的基础窦状卵泡数、基础孕激素、卵泡数、可移植胚胎数等呈正相关关系，体外实验提示MALAT1可能通过ERK/MAPK信号通路调控细胞周期蛋白抑制因子p21和p53的表达进而影响卵巢颗粒细胞增殖。本项目通过收集EMT不孕患者卵泡液颗粒细胞，并利用KGN颗粒细胞系及小鼠模型，探讨MALAT1在颗粒细胞类固醇分泌、增殖及凋亡、卵泡发育及卵巢反应中的作用及机制，构建MALAT1-miRNA-mRNA调控网络，为保护女性生育功能提供依据。

子宫内膜异位症（EMT）是育龄妇女常见的慢性疾病，表现为进行性痛经、月经异常、不孕等。体外受精-胚胎移植是EMT的有效助孕方法，然而EMT造成卵巢储备功能下降影响了超促排卵的效果。本项目研究发现：在EMT不孕患者卵泡液颗粒细胞中lncRNA MALAT1表达水平明显下调。与轻度内异症（I-II期）患者相比，MALAT1在重度内异症（III-IV期）患者颗粒细胞中的表达水平更低。临床数据分析显示MALAT1与AFC、基础孕激素水水平、获卵数、成熟卵泡数、正常受精率、可移植胚胎数及优质胚胎数呈正相关关系；与患者年龄呈负相关。体外研究提示，沉默MALAT1后明显抑制颗粒细胞的增殖，沉默MALAT1后异常激活ERK/MAPK信号通路，异常激活的ERK/MAPK通路能稳定细胞周期蛋白依赖性激酶抑制因子p21、p53，导致p21、p53蛋白水平上调，cyclin D1、CDK2表达水平下调，从而阻滞了细胞周期进展，抑制颗粒细胞的增殖。MALAT1还可能通过调控ACSL4、FTH1、SLC7A11的转录及翻译从而参与细胞铁死亡的调节，进而参与调控内异症患者的卵巢功能。然而，MALAT1可能不影响卵巢颗粒细胞的凋亡，亦不影响颗粒细胞芳香化酶的表达及甾体激素的水平。此外，本课题组还拓展了其他lncRNA参与卵巢功能调控的机制研究。lncRNA NEAT1在子宫内膜异位症不孕患者的卵泡液颗粒细胞中表达下调，NEAT1可能通过影响颗粒细胞增殖影响卵巢功能。卵巢储备功能下降是内异症患者生育力下降的重要原因，lncRNA PVT1在生化期卵巢功能不全（bPOI）患者的表达显著下降，PVT1可能通过调节颗粒细胞的增殖和自噬破坏卵泡的形成和成熟，其可能成为bPOI的一种新的诊断标志物。本项目揭示了内异症不孕患者卵巢储备功能减退机制，为改善不孕患者辅助生殖治疗结局提供实验研究的基础。