PGE2和PGF2α对奶牛子宫内膜上皮细胞促增殖作用及其机制研究

Prostaglandins play important roles in female reproductive processes that include ovulation, fertilization, implantation, decidualization and placentation through their specific receptors. However, it is unclear that the influence of prostaglandins in endometrial cell proliferation and its mechanism of action. In this project, bovine endometrium is used as the main research object, which illuminates the effects of prostaglandin E2 and prostaglandin F2α on acceleration of the endometrial epithelial cells proliferation and growth factors including VEGF, CTGF, TGF and IL-8 secretion in bovine endometrial epithelial cells. Meanwhile, the regulatory roles of PI3K/Akt signaling in endometrial epithelial cell proliferation were analyzed by using the methods of cell and tissue culture in vitro, real-time cellular analysis, patch clamp recording technique, flow cytometry, ELISA, real-time quantitative PCR, Western blot and immunofluorescence staining. The results will illuminate the mechanism of prostaglandin E2 and prostaglandin F2α promoting the endometrial epithelial cells proliferation and provide theoretical and experimental basis for treatment of bovine reproductive system diseases using prostaglandins clinically.

前列腺素类化合物作为局部激素通过激活其特定受体在排卵、受精以及胚胎发育等生殖过程中发挥重要作用，但尚不清楚前列腺素类化合物对子宫内膜上皮细胞增殖的影响及其作用机制。本项目以奶牛子宫内膜作为研究对象，采用体外细胞及组织培养、实时无标记细胞检测、膜片钳技术、流式细胞术、ELISA、荧光定量PCR、Western blot、免疫荧光染色等实验技术，研究PGE2和PGF2α对子宫内膜上皮细胞增殖的促进作用；研究PGE2和PGF2α对奶牛子宫内膜上皮细胞合成分泌VEGF、CTGF、TGF和IL-8等细胞组织生长相关因子的作用；分析PGE2和PGF2α是否通过子宫内膜上皮细胞PI3K/Akt信号转导通路的激活对细胞增殖进行调控。阐明PGE2和PGF2α促进子宫内膜上皮细胞增殖的机制，从而为临床上合理应用前列腺素类药物防治奶牛繁殖障碍疾病提供理论和实验依据。

前列腺素类化合物作为局部激素通过激活其特定受体在排卵、受精以及胚胎发育等生殖过程中发挥重要作用。但尚不清楚前列腺素类化合物对子宫内膜上皮细胞增殖的影响及其作用机制。本研究已经按照预定计划完成了规定的任务，主要在前列腺素F2α（prostaglandin F2α，PGF2α）和前列腺素E2（prostaglandin E2，PGE2）对奶牛子宫内膜上皮细胞（bovine endometrial epithelial cells，bEECs）促增殖作用机制的研究上取得了一定突破和成果。本项目研究发现，PGF2α-PTGFR和PGE2-PTGER2信号通路激活可通过对细胞周期的调控，诱导奶牛子宫内膜上皮细胞的增殖，从而促进子宫内膜组织的修复，进而发挥维持子宫微环境稳态的作用。此外，本课题组通过拓展性研究发现，PGE2参与NLRP3介导的小鼠肌成纤维细胞对巨噬细胞分泌活动的影响，进而促进小鼠创伤修复的进程。因此，在奶牛子宫内膜炎的防治工作中，以PGE2、PGF2α及其结构类似物为基础的促组织修复制剂的开发，对该病的防治具有较高的潜在应用价值。