针刺上调HLA-G/E的表达调节母胎免疫耐受促进胚胎着床的分子机制研究

It is a tough problem that the implantation rate of in vitro fertilization-embryo transfer (IVF-ET) is relatively low. The latest result of meta analysis about acupuncture and IVF has showed that acupuncture can significantly increase the success rate of IVF-ET. However, the mechanism is still unclear. Basing on the previous animal experiment, this project will use the method of controlled ovarian hyperstimulation to make the rat model of embryo implantation obstacle that is consistent with the process in IVF-ET, observe the effect of acupuncture on the embryo implantation and pregnancy rate, and detect in blood serum and maternal-fetal interface the level of estrogen, progestogen and their receptors, the counter part of HLA-G/E, immunocytes, and cytokines using immunohistochemistry，enzyme linked immunosorbent assay, real time polymerase chain reaction, western blot and fluorescence-activated cell sorting. The objective is to verify that acupuncture may directly or indirectly up-regulate the expression of HLA-G/E, improve the maternal-fetal immune tolerance, promote embryo implantation and elevate pregnancy rate. Elucidating the molecular mechanism of acupuncture promoting embryo implantation can provide theory evidence for the combination of acupuncture and modern assisted reproductive technology, and establish the foundation of the application and extension of acupuncture in reproduction field.

胚胎着床率较低一直是体外受精-胚胎移植（IVF-ET）技术中的难题。最新的Meta分析结果表明针刺能显著提高IVF-ET的妊娠率，但其作用机理仍不清楚。本项目在前期动物实验的基础上，采用与临床IVF-ET中相一致的控制性超排卵方法造成胚胎着床障碍大鼠模型，以传统中医理论为指导，观察针刺对此模型鼠胚胎着床和妊娠率的影响；应用免疫组化、ELISA、Western blot、实时荧光定量PCR及FACS等现代细胞生物学和分子生物学技术，系统检测大鼠血清、母胎界面中雌、孕激素及其受体、HLA-G/E对应物、免疫细胞及细胞因子等指标的变化，以期验证："针刺可能通过直接或间接上调HLA-G/E(对应物)的表达，改善母胎免疫耐受而实现其促进胚胎着床、提高妊娠率的作用"，初步阐明针刺促进胚胎着床的分子机制，为针刺与IVF-ET等辅助生殖技术相结合提高妊娠率提供理论依据，为针刺在生殖领域的推广应用奠定基础。

体外受精-胚胎移植（IVF-ET）技术中，胚胎着床率较低一直是难题。最新的Meta分析结果表明针刺能显著提高IVF-ET的妊娠率，但其作用机理仍不清楚。本项目在前期动物实验的基础上，采用与临床IVF-ET中相一致的控制性超排卵方法造成胚胎着床障碍大鼠模型，以传统中医理论为指导，观察了针刺对此模型鼠胚胎着床和妊娠率的影响；应用ELISA、Western blot、实时荧光定量PCR等现代细胞生物学和分子生物学技术，系统检测聊了大鼠血清、母胎界面中雌、孕激素及其受体、HLA-G/E对应物、免疫细胞及细胞因子等指标的变化，验证了：“针刺通过直接或间接上调HLA-G/E(对应物)的表达，改善母胎免疫耐受而实现其促进胚胎着床、提高妊娠率的作用”，初步阐明了针刺促进胚胎着床的分子机制，为针刺与IVF-ET等辅助生殖技术相结合提高妊娠率提供了理论依据，为针刺在生殖领域的推广应用奠定基础。