TWEAK对山羊uNK细胞分化和功能的调节作用及其机制

妊娠早期子宫内膜基质部子宫NK细胞（uNK）表型及功能变化与胚胎能否成功植入以及妊娠确立密切相关，但其分化及功能调节机制尚不十分清楚。肿瘤坏死因子样凋亡微弱诱导剂（TWEAK）在机体正常生理条件下免疫动态平衡的维持以及炎症状态下免疫功能调节中具有重要作用。本研究应用miRNA芯片分析技术、双荧光素酶检测系统等筛选靶向调节山羊uNK细胞中TWEAK的miRNA，进一步通过荧光定量RT-PCR、Western blot、流式细胞术、基因干扰及过表达等方法检测miRNA对uNK细胞中TWEAK转录后水平的表达调控机制及其在山羊妊娠早期uNK细胞分化及分泌表达中的调节作用，以及不同类型胚胎和激素对该作用的影响，阐明山羊妊娠早期uNK细胞表型及免疫功能的TWEAK分子调控机制。本研究对全面了解妊娠免疫，提高胚胎移植成功率和家畜繁殖率，以及生殖道抗感染免疫机制等均具有重要的理论意义和潜在的应用价值。

The regulation of uterine natural killer (uNK) cells of phenotypes and functional transition has been closely associated with implantation failure and reproduction development. However, the comprehensive functions of uNK cell in early pregnancy still remain unclear. Tumor necrosis factor-like weak inducer of apotisis (TWEAK) is an important regulator of the development, homeostasis, and function of the immune system. In the present study, the miRNA target TWEAK was detected by using miRNA microarray and dual-luciferase assay. Furthermore, the regulatory mechanisms of TWEAK expression post-transcriptionally and its role in phenotypes regulation and secretory functions of uNK cells, as well as the effects of embryo types and steroids on the above regulation were detected using quantitative RT-PCR, western blot, flow cytometric assay, and RNAi assay. This study will illustrate TWEAK regulation mechanisms of phenotypes and immune function of uNK cell. Moreover, this study may be pivotal significance to improve the successful rate of domestic animal reproduction, transplantation immune, and provide new mechanistic insights into anti-infection immunity in reproductive tracts.