多囊卵巢综合征痰湿内生的细胞脂代谢稳态机制

The pathogenesis of polycystic ovarian syndrome (PCOS) is involved with deficient Zang Qi and stagnated phlegm-dampness in the traditional Chinese medicine, but the cytological mechanism is still unclear in the western medicine. The endogenetic phlegm-dampness can be induced by abnormal transportation and transformation in the traditional Chinese medicine. The accumulated lipid droplets can be caused by lipid metabolic imbalance in the western medcine. What is known above with different aspects can be integrated for a confluence of theories. In our previous studies, abnormal very low-density lipoproteins (VLDL) and high-density lipoproteins (HDL) were observed generally in the patients with PCOS. We have invented the MEDLiPO system (an approved patent) for lipoprotein separation and quantification, and found a special HDL subclass: HDL(A). Apolipoprotein E is rich in either HDL(A) or VLDL particles. A low level of HDL(A) and a high level of VLDL were found in the patients with PCOS. Therefore, the metabolic imbalance of these lipoproteins might be associated with endogenic phlegm-dampness in PCOS. In this study, the effects of HDL(A) and VLDL on rat ovarian follicle cells will be observed in vitro. Research contents include: 1. Receptor-mediated transfer of lipoproteins into cells. 2. Formation and mobilization of intracellular fatty acids and lipid droplets. 3. Genetic expression of steroidogenetic factors and the secretion of gonadal hormones. The program is designed to study the mechanism of cellular lipid homeostasis of endogenic phlegm-dampness in PCOS, providing experimental evidence for clinical treatment on PCOS by lipid regulation and sputum analysis.

多囊卵巢综合征（PCOS）的病机在中医理论认为主要有脏气虚损，痰壅湿阻，在西医细胞学机制尚未阐明。中医运化失调而痰湿内生，西医脂代谢失衡而脂滴蓄积，两种角度的认识可以汇通或融合。我们前期研究发现：PCOS患者普遍存在血清极低密度脂蛋白（VLDL）和高密度脂蛋白（HDL）含量异常。我们发明MEDLiPO系统的专利技术，研究发现HDL有一种特殊的亚类-HDL(A)。HDL(A)和VLDL在结构上都富含载脂蛋白E，它们在PCOS患者中的含量是此消彼长，其代谢失衡可能和PCOS痰湿内生有关。本项目拟观察HDL(A)和VLDL对大鼠卵泡细胞的干预作用，研究细胞经受体介导途径转运脂蛋白；细胞内脂肪酸、脂滴的形成和动员；甾体合成相关因子的基因表达和性激素的分泌。旨在从细胞脂代谢稳态的角度研究PCOS痰湿内生的机制，进而为临床通过调节血脂和从痰论治PCOS提供实验依据。

多囊卵巢综合征（PCOS）的病机在中医理论认为脏气虚损，痰壅湿阻。西医则认为脂代谢和生殖有密切的关系，但是相关脂蛋白的研究甚少。我们围绕细胞内脂滴来研究和痰湿内生的关系，探讨脂蛋白参与生殖微环境脂质转运和转化的机制。本项目选择OVCAR3细胞作为研究对象，建立油酸负荷的细胞内脂滴模型。油酸处理细胞后胞内微粒体甘油三酯转运蛋白（MTTP）显著上调，提示脂质应激性上调MTTP利于脂质外排至胞外。油酸干预三磷酸腺苷结合盒转运子A1（ABCA1）表达下调，提示细胞内脂质通过高密度脂蛋白（HDL）/ABCA1外流的通路受抑制。MTTP敲减后ABCA1表达上调，说明脂代谢基因调控存在交互作用机制。过表达MTTP有可能逆转脂质诱导的细胞脂滴形成。脂蛋白干预实验表明，极低密度脂蛋白（VLDL）促进OVCAR3细胞雄激素的分泌，HDL则抑制雄激素分泌，两者作用相反。由于卵泡屏障作用，卵泡液（FF）中富含小颗粒的VHDL。针对VHDL的检测技术存在明显的不确定性，我们围绕VHDL开展了艰难的方法学实验研究，包括改良梯度凝胶电泳和脂染色定量分析，优化MEDLiPO血脂检测体系，定制BeneScan扫描和分析系统，已申请发明专利。该系统成功应用于VHDL颗粒重塑的体外实验。就鉴定VHDL的工作花费了1年的时间，最终取得突破。发现PCOS患者FF中VHDL含量降低，新生儿VHDL颗粒重塑处于极其活跃的动态过程。先后发表论文和参加学术会议交流，本项目研究结果在本领域受到关注。