基于《内经》“七七”理论的高龄IVF卵泡液生物标志物筛选及经方干预研究

According to the research results of our former seven National Nature Science Fund subjects, this project aims to explore the scientific connotation of “NEIJING” “Qi-Qi”theory, with the technology of “reverse engineering” as the research method. Starting with follicular fluid on the DOPU of both the “Wu-Qi” “Liu-Qi” aged patients of Kindey-Qi deficiency and the “San-Qi” “Si-Qi” normal ones in the growth stage with IVF-ET,analysis of metabolism fingerprint spectrum of the follicle fluid in the “Wu-Qi” “Liu-Qi” aged patients of Kindey-Qi deficiency and the “San-Qi”“Si-Qi” normal ones in the growth stage before and after taking classical Chinese medicine is done by UPLC-Q-TOF/MS, GC-MS/NMR and biological information calculation method, to recognize and determine the characteristic metabolites compounds closely related to the aged Kindey-Qi deficiency syndrome and the effect of classical prescription.It will help to find the biology markers of infertility due to Kidney deficiency in the aged, and provide theoretical and experimental basis for scientific prediction, diagnosis and discussion of kidney deficiency syndrome of the aged. Furthermore, it is expected to be applied to clinical diagnosis, treatment, prognoses analysis and drug development, and give impetus to the development of “Qi-Qi” Theory which is based on follicular fluid metabonomics.

项目在前七项国家自然基金课题研究的基础上，以挖掘《内经》“七七”理论的科学内涵为目标，以代谢组学中“反向工程学”的技术思路为研究方法，以体外受精-胚胎移植技术为平台，以“五七”、“六七”高龄肾气虚及“三七”、“四七”生育阶段正常女性取卵日的卵泡液为切入点，利用UPLC-Q-TOF/MS、GC-MS/NMR多种互补技术及生物信息计算手段，获取“五七”、“六七”高龄肾气虚及“三七”、“四七”生育阶段正常女性和经典中药干预后的卵泡液代谢物指纹图谱，对各组卵泡液代谢图谱表达进行全景式分析，辨识并确定与高龄肾气虚证发生和经方作用机理密切相关的代谢物特征性化合物；有助于发现高龄肾气虚不孕的生物学标记物，为科学预测和诊断高龄肾气虚证、探讨其证候本质提供理论和实验依据，以期应用于临床诊断、治疗、预后分析和药物研发，力争在卵泡液代谢组学层面丰富发展“七七”理论。

项目以深入挖掘《内经》“七七”理论的科学内涵为目标，聚焦高龄不孕女性，依托IVF-ET技术，以代谢组学中“反向工程学”的技术思路为研究方法，以“五七”、“六七”高龄肾气虚及“三七”、“四七”生育阶段正常女性取卵日的卵泡液为切入点，利用UPLC-QTOF/MS、GC-MS/NMR多种互补技术和分析平台，获取“五七”、“六七”高龄肾气虚及“三七”、“四七”生育阶段正常女性和补肾中药干预后的卵泡液代谢物指纹图谱，对各组卵泡液代谢图谱表达进行全景式分析，辨识并确定与高龄肾气虚证发生和补肾中药作用机理密切相关的代谢物特征性化合物。研究结果表明：激素类、肉碱类、胆碱类、脂肪酸类物质与女性的衰老及卵母细胞质量密切相关，推测上述物质可能是高龄肾气虚证潜在的生物标志物。补肾中药发挥作用的机制可能与类固醇激素合成通路、甘油磷脂代谢通路、脂肪酸氧化代谢通路、氨基酸代谢及能量代谢通路等有关，而补肾中药发挥作用的靶点则是该通路上与代谢物相关的蛋白或蛋白酶。本研究为科学预测和诊断高龄肾气虚证、探讨其证候本质提供理论和实验依据，从卵泡液代谢组学层面探寻高龄肾气虚型不孕潜在的证候要素及补肾中药的作用靶点，在丰富和发展《内经》“七七”理论的同时，为高龄试管婴儿患者探索更优化的治疗方案提供有益的尝试。

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