补肾法、疏肝法诱发排卵与PR调节卵泡破裂的关系

Previous researches demonstrate that Bushen Treatment and Shugan Treatment sequential therapy can increase ovulation rate in mice preprocessed with Gonadotropin, and regulate the correlation factor inducing ovulation. The number increase may be relevant to oocyte release with follicular rupture as a prerequisite in ovulation. PR is essential for follicular rupture. Mice null for PR (PRKO) fail to release the oocytes. Ovulation of rats or mice were completely inhibited when the progesterone antagonist mifepristone (RU486) were given. The factors relavant to the rupture and regulated by PR involve peroxisome proliferator-activated receptor γ(PPARγ), a disintegrin and metalloproteinase domain with thrombospondin motif (ADAMTS-1), a disintegrin and metalloprotease（ADAM 8）, and endothelin-2(ET-2). This project aims to explore the relationship between Bushen & Shugan treatment-induced ovulation and PR-regulated factors, and the differences and similarities in their functional mechanisms by detecting the dynamic effect of Bushentiaojing recipe and Xiaoyao pill on those factors in the ovary of the mice preprocessed with Gonadotropin and RU486 (PR inhibitor)intervened, and in the ovum cultured in vitro, which is of great significance for treating anovulatory disorders with Bushen and Shugan herbal sequential therapy, explaining mechanism of regualting ovulation, enriching the theory of "Kidney governing reproduction" and "Liver being the congenital of women's reproduction" and perfecting the reproduction theory of traditional Chinese medicine.

前期研究及预试验表明，补肾法、疏肝法均可提高促性腺激素预处理小鼠排卵率，并可调节诱发排卵的相关因子。卵泡破裂是排卵的必要条件。PR是排卵所必需的，PR基因敲除小鼠卵泡不能破裂，孕激素拮抗剂RU486可抑制大鼠、小鼠排卵。与卵泡破裂有关并受PR调节的因子有过氧化物酶体增殖剂激活型受体(PPAR)γ、去整合素-金属蛋白酶家族的ADAMTS-1、ADAM 8以及内皮素-2(ET-2)。本项目以补肾调经方、逍遥丸分别为补肾法、疏肝法的代表方，拟通过研究补肾调经方、逍遥丸对促性腺激素预处理小鼠卵巢、RU486干预的促性腺激素预处理小鼠卵巢以及体外培养的小鼠卵泡中上述因子的动态影响，探讨补肾法、疏肝法诱发排卵与PR调节卵泡破裂的关系及其作用机制之异同。这对揭示补肾、疏肝中药治疗排卵障碍性疾病及改善女子氤氲状态的作用机制，丰富"肾主生殖"、"女子以肝为先天"的理论内涵，完善中医生殖理论具有重要意义。

背景：排卵障碍是女性不孕症的主要原因之一，而卵泡发育成熟是排卵的前提、排卵孔的形成是卵母细胞排出的关键。排卵障碍的基本中医病机是肾虚、肝郁，补肾法、疏肝法可促进卵泡发育、诱发排卵，临床疗效肯定，中药诱发排卵机制值得研究。.主要研究内容：研究补肾调经方、逍遥丸对促性腺激素预处理小鼠卵巢及体外培养的小鼠卵泡中PR及ADAMTS-1、ADAM 8、ET-2、PPAR γ、HIF1-α、ET-2、GDF-9、BMP-15 mRNA及其蛋白表达的影响，探讨补肾法、疏肝法诱发排卵与PR及其相关因子调节卵泡破裂诱发排卵的关系及其机制之异同。.重要结果：补肾调经方、逍遥丸均可使促性腺激素预处理小鼠卵巢排卵数目增多，两方均可上调小鼠排卵前卵巢PR mRNA及其调节因子和卵母细胞分泌因子GDF-9、BMP-15 mRNA蛋白的表达，促进排卵斑形成、卵泡壁破裂而诱发了排卵。两法作用机制亦有不同，补肾法小鼠卵巢ADAMTS-1、PPARγ、ET-2mRNA及其蛋白的表达高峰在排卵时（注射hCG后12h），疏肝法卵巢ADAMTS-1、PPARγ mRNA及其蛋白的表达在排卵前（注射hCG后8h）；两法对小鼠卵巢HIF1-αmRNA及其蛋白的表达影响无明显差别。两方含药血清可提高体外培养小鼠窦前卵泡的存活率，补肾法优于疏肝法；补肾法促进卵泡ADAMTS-1、ADAM-8、PPARγmRNA及其蛋白表达优于疏肝组，据“肾主生殖、为先天之本”推测与补肾法促进卵泡发育作用较强有关。补肾调经方作用机制与阴精盛助卵泡发育、阳气足使阴阳转化顺利有关；逍遥丸与血足精充促卵泡发育、疏肝理气助阴阳转化顺利有关。.科学意义：补肾法和疏肝法均有诱发排卵作用，其机制有同有异。为临床使用补肾法、疏肝法治疗排卵障碍性疾病提供生理基础，阐释中医改善女子氤氲状态的作用机制以及“肾藏精、主生殖、为先天之本”、“女子以肝为先天”、“肝主疏泄”等理论内涵。