补肾固冲法诱导DCreg介导URSA母胎免疫耐受作用与机制研究

Unexplained recurrent spontaneous abortion (URSA) harms women's reproductive health seriously, and exploring effective treatment is a hotspot in the field of obstetric and gynecology. In traditional Chinese medicine (TCM), it was believed that "kidney governing reproduction" and the method of "invigorating kidney and consolidating chong vessel" is the basic principle of URSA treatment. However, the initial target of the method has not been fully clarified. Maternofetal immunotolerance mediated by T cells plays a crucial role in the success of pregnancy. There is disequilibrium of T-cell subsets in URSA; moreover, it was found in our preliminary study that the disequilibrium is closely related to the abortion rate. However, what is the initial factor which causes the disequilibrium of T-cell subsets to destroy the maternofetal immunotolerance is unclear now. Regulatory dendritic cell (DCreg) is a new dendritic subset which was found recently. DCreg plays a crutial role in inducing immunotolerance by activating and regulating T cells. Interestingly, it was found in our preliminary experiment that the number and function of DCreg changed in URSA mice; furthermore, the data showed that the method of "invigorating kidney and consolidating chong vessel" had the regulating effect both on the quality and the function of DCreg in vitro. Therefore, it was hypothesized that the inducing effect of the method "invigorating kidney and consolidating chong vessel" mediates maternofetal immunotolerance in URSA by inducing DCreg. In this study, URSA mice model will be uesd, meanwhile, the number and function of DCreg as well as T-cell subsets in URSA mice before and after the treatment of "invigorating kidney and consolidating chong vessel" will be observed. The result of the study will demonstrate the inducing effect of invigorating kidney and consolidating chong vessel on maternofetal immunotolerance and elucidate the underlying mechanism. Moreover, the findings of the study will enrich the scientific meaning of "kidney governing reproduction" theory in TCM and provide novel theraputical target and method for clinics.

原因不明复发性流产（URSA）严重危害妇女生殖健康，其有效治疗方法的发掘是研究的热点。以"肾主生殖"为理论基础的补肾固冲法是URSA基本治疗法则，已证实该法能逆转URSA母胎免疫耐受紊乱的T细胞亚群失衡现象，但该法诱导母胎免疫耐受的关键始动靶点尚不清楚。本研究根据调节性树突状细胞(DCreg)调节T细胞增殖分化在诱导免疫耐受中发挥关键作用的前沿理论，在预实验发现URSA小鼠存在DCreg数量与功能改变、补肾固冲法能体外调节DCreg数量与功能的基础上，拟通过建立URSA小鼠模型，从细胞、分子、基因水平，观察补肾固冲法作用前后模型小鼠DCreg数量及其表面共刺激分子、MHCⅡ类分子、T细胞亚群分化信号分子表达变化，同步观察T细胞状态及妊娠结局，明确补肾固冲法诱导DCreg调节T细胞亚群分化介导URSA母胎免疫耐受的作用与分子机制，丰富中医学"肾主生殖"理论科学内涵，为治疗提供新的靶点与途径。

原因不明复发性流产（URSA）严重危害妇女生殖健康，其有效治疗方法的发掘是研究的热点。以“肾主生殖”为理论基础的补肾固冲法是URSA基本治疗法则，已证实该法能逆转URSA母胎免疫耐受紊乱的T细胞亚群失衡现象，但该法诱导母胎免疫耐受的关键始动靶点尚不清楚。本研究在国家自然科学基金面上项目“补肾固冲法诱导DCreg介导URSA母胎免疫耐受作用与机制研究（81373670）”资助下，重点探讨了树突状细胞（DC）亚群与功能变化在URSA中的作用及补肾固冲法靶向干预机制。研究内容及创新点包括：1. URSA患者及模型动物存在DC亚群及功能改变。pDC比例降低、cDC比例升高、pDC/cDC比率降低。DC表面MHC-II分子、共刺激分子（CD80/CD86）及功能性分子（CD275/CD205/Tim3）表达升高，CD274、33D1表达降低；细胞因子IL-6、IL-12、IL-23分泌增多，IL-10、TGF-β分泌减少。2. 补肾固冲法能有效降低URSA小鼠胚胎吸收率，对妊娠具有保护作用。3. 补肾固冲法能够逆转URSA小鼠及患者DC亚群偏移状态，降低DC表面MHC-II分子、共刺激分子及功能性分子表达，升高CD274、33D1表达，诱导母胎免疫耐受，治疗URSA。4. 补肾固冲法作用后，小鼠E2-2表达升高，STAT5、ID-2表达降低，提示JAK/STAT信号通路是中药干预DC分化与功能治疗URSA的关键机制之一，表观遗传机制可能在此过程中发挥着重要调控作用，其深入探讨有望揭示URSA发生的表观遗传免疫调控机制，进一步丰富中医学“肾主生殖”理论的科学内涵。. 研究结果进一步阐释了URSA发生的免疫学机制，为补肾固冲法诱导母胎免疫耐受提供了实验依据。本研究发表SCI收录论文5篇，中文核心期刊发表论文6篇，参加会议交流6次（国际会议交流2次、国内会议交流4次），获得山东省科技进步二等奖及山东省中医药科技一等奖各1项，培养博士后1名、硕士研究生5名；并在此基础上获得了国家中组部留学归国人员择优资助项目、中国博士后基金特别资助的支持。部分工作正在投稿或审稿中，后续工作将及时向基金委汇报。