基于MHC的补肾复方对COH小鼠子宫内膜容受性调控机制研究

The clinical application of In vitro fertilization-embryo transplantation (IVF-ET) is more and more popular. Endometrial receptivity is the key in IVF-ET success. Modern medicine takes pregnancy as allotransplantation, in which major histocompatibility complex(MHC) is essentially important. Since it controls and regulates the endometrial receptivity, it shows a close relationship with pregnancy. The research team realizes that TCM theory of “kidney governing reproduction as the dominator of essence storage” is related to endometrial receptivity. Thus, the purpose of the study is to explore if “Shou Tao Wan”(STW)—a classic formula of kidney-tonifying for miscarriage prevention, could increase endometrial receptivity by regulating MHC expression. The study is supposed to establish a pattern of COH mice model combined with primary cultured endometrial cell from COH mice model, with luminex, Q-PCR, Western blot, and so on, to detect influence of Shou Tai Wan on endometrial histomorphology at window stage for inplantation, MHC and its relevant factors expression in endometrium and peripheralblood cells. Meantime, the influence of STW on endometrial MHC and its relevant factors would be measured, to analyze that STW could increase endometrial receptivity by regulating MHC. We would try to reveal the molecular regulation mechanism of kidney-tonifying formula on improving endometrial receptivity from the aspect of MHC, therefore to enrich the TCM theory of “kidney governing reproductive as the dominator of essence storage” and provide evidences of integrating kidney-tonifying formula in IVF-ET.

IVF-ET临床应用日益增多，子宫内膜容受性是IVF-ET成功与否的重要环节。妊娠类比同种异体移植，而MHC分子在同种异体移植中至关重要，其与妊娠密切相关，调控子宫内膜容受性。课题组认为“肾主生殖，为封藏之本”的理论与内膜容受性相关，鉴此，补肾安胎经典方“寿胎丸”能否通过调节MHC分子提高子宫内膜容受性？课题拟通过体内（COH模型小鼠）—体外（原代培养COH模型小鼠子宫内膜细胞）联合，采用液相芯片、Q-PCR、WB等方法，观察寿胎丸对COH模型小鼠种植窗期子宫内膜组织形态、内膜及外周血细胞MHC分子及其相关因子的影响，观察寿胎丸对子宫内膜细胞MHC分子及其相关因子的影响，分析寿胎丸能否通过调节MHC分子，从而提高子宫内膜容受性。从 MHC角度，在分子水平，揭示补肾复方改善COH子宫内膜容受性的调控机制。丰富中医“肾主生殖，为封藏之本”的客观化内涵，为IVF-ET中应用补肾复方提供依据。

IVF-ET临床应用日益增多，子宫内膜容受性是IVF-ET成功与否的重要环节。现代认为妊娠类比同种异体移植，MHC分子在同种异体移植中至关重要，调控子宫内膜容受性。课题组认为“肾为封藏之本”的理论与内膜容受性相关，鉴此，补肾安胎经典方“寿胎丸”能否通过调节MHC分子提高子宫内膜容受性？本研究建立COH大/小鼠，以寿胎丸予以干预COH鼠，制备子宫内膜细胞及含寿胎丸血清，以含寿胎丸血清干预子宫内膜细胞，采用液相芯片、免疫组化、Q-PCR等方法，检测子宫内膜细胞MHC分子及其相关因子的水平。在体实验结果：与COH模型组比较，寿胎丸高/中/低剂量组MHC-1、JNK2、E-cadherin、P-Cadherin、IL-1b、IL-4、IL-10、IP-10、VEGF水平及CD8+T细胞含量明显升高；IL-1a、IL-2、IL-17、TNF-a、IFN-y水平及CD4+T细胞含量明显降低；子宫内膜LIF、ITGAV、ITGB3、ITGA5、VEGF、VEGFR-1蛋白Area/IOD明显增加；子宫ITGAV、LIF、ITGB3、ITGA5、VEGF及VEGFR-1mRNA水平明显升高；MHC-2、E2、P水平及CD3+T细胞含量无变化。体外实验结果：与COH血清组比较，寿胎丸高/中/低剂量血清组MHC-1、E-cadherin、P-Cadherin、IL-4、IL-6、VEGF水平明显升高，IFNγ、TNFα、IL-2水平明显下降，子宫内膜细胞LIF、ITGAV、ITGβ3、ITGβ5mRNA水平明显升高。E2、P、MHC-2水平无变化。研究提示，寿胎丸可能通过调控MHC-1水平、CD4+T/CD8+T细胞比例，调节JNK2、E-cadherin、P-Cadherin、IL-1b、IL-4、IL-10、IP-10、VEGF、IL-1a、IL-2、IL-17、TNF-a、IFN-y、IL-6水平，影响子宫内膜LIF、ITGAV、ITGA5、ITGB3、VEGF、VEGFR-1蛋白及mRNA水平，而提高COH子宫内膜容受性。研究从MHC角度，在分子水平，揭示补肾复方改善COH子宫内膜容受性的调控机制。丰富中医“肾为封藏之本”的客观化内涵，为IVF-ET中应用补肾复方提供依据。培养省/局学术技术带头人后备人选各1人，硕士6名，参加学术会议6人次，发表学术论文5篇，基本完成预期计划。