基于体外受精-胚胎移植卵泡液代谢组学的肾主生殖理论研究

According to National Nature Science Fund subject researchs in the past, Chinese medicine for kidney-tonify can improve the quality of oocyte from epigenetics, genomics and proteomics, but preliminary studies cann't properly reflect the changes of oocyte metabolic state. Hereby, this project aims to explore the theory of Kidney Governing Reproduction Theory, with the technology of 'reverse engineering' as research method. Starting with follicular fluid of Kidney-Yin deficiency, Kidney-Yang deficiency, Kidney-Qi deficiency and normal group of patients with IVF-ET, analysis of metabolism finger-print of Kidney-Yin deficiency, Kidney-Yang deficiency, Kidney-Qi deficiency and normal group of patients before and after taking Chinese medicine for kidney-tonify 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 Kidney deficiency and the mechanism of Chinese medicines for kidney-tonify. It will be able to help find the biology markers of infertility due to Kidney deficiency, and provide theoretical and experimental basis for scientific prediction, diagnosis and discussion of kidney deficiency syndrome. Furthermore, it is expected to be applied to clinical diagnosis, treatment, prognoses analysis and drug development, and give impetus to the development of Kidney Governing Reproduction Theory based on follicular fluid metabonomics.

课题组前四项国家自然基金课题从表观遗传学、基因组学、蛋白质组学等角度研究证实，补肾中药可改善卵细胞质量。但前期研究不能很好地反映卵母细胞代谢状态的变化。基于此，本项目以挖掘"肾主生殖"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，以肾阴虚、肾阳虚、肾气虚及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，获取正常人及肾阴虚、肾阳虚、肾气虚证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，辨识并确定与肾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；有助于发现肾虚不孕的生物学标记物，为科学预测和诊断肾虚证、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、预后分析和药物开发，力争在卵泡液代谢组学层面丰富发展"肾主生殖"理论。

项目以挖掘“肾主生殖”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，以肾虚、血瘀、痰湿及正常女性取卵日的卵泡液为切入点，利用代谢组学技术，获取正常人及肾虚、血瘀，痰湿等中医经典不孕证型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，辨识并确定与肾虚、血瘀、痰湿证发生作用机理密切相关的代谢物特征性化合物；研究结果表明：肾虚不孕标志物主要涉及到脂类、核苷酸类、蛋白类；痰湿不孕生物标识主要涉及磷脂脂类、类固醇类以及氨基酸类等；血瘀型不孕症生物标志物主要分为脂类、核苷酸类、蛋白降解片段及其它等四大类，项目发表论文18篇，其中SCI收录1篇，目前在投SCI文章4篇，培养硕士研究生3名，申请国家发明专利3项，在卵泡液代谢组学层面丰富发展“肾主生殖”理论。