基于中医窠囊理论探讨桂枝茯苓丸改善PCOS-IR大鼠排卵障碍的研究

PCOS is a common and refractory gynecologic disease, of which ovulation disorder is the main symptom impairing the reproductive health of females of child-bearing age to a great extent. Based on the theory of ‘the Kenang was induced by phlegm intermingling static blood’ in Danxi’s Mastery of Medicine, the study acknowledges that phlegm and static blood are the basic pathogenesis of both PCOS common clinical performances and ovarian morphologic changes. Starting with intestinal flora and regulation of ovarian autophagy signaling pathway, the research will discuss whether to remove stasis and phlegm through the fang guizhi poria bolus could adjust intestinal flora, increase intestinal permeability and have a regulating role for inflammatory agents Sequently, the insulin sensitivity of PCOS-IR might be improved to inhibit granular cells’ phagocytosis by activating PI3K/Akt/mTOR signaling pathway. Finally ovarian follicle maturation might be promoted that would play a part in the treatment of PCOS-IR. This study will reveal the mechanism of the treatment of PCOS-IR ovulation disorder by using the fang guizhi tuckahoe pill, provide a theoretical basis for the treatment of PCOS-IR, improve the clinical therapeutic effect of PCOS and augment the clinical application of the fang guizhi tuckahoe pill.

PCOS为妇科常见疑难疾病，排卵障碍是其主要症状及危害，极大影响了生育期妇女生殖健康。 本研究基于《丹溪心法》“痰挟瘀血，遂成窠囊”理论，从PCOS常见临床表现及卵巢形态学改变认识到痰瘀互结为基本病机，从肠道菌群及调控卵巢自噬信号通路入手，探讨祛瘀化痰经方桂枝茯苓丸是否通过调节肠道菌群、改善肠道通透性、调控炎性因子，进而提高PCOS-IR的胰岛素敏感性，激活PI3K/Akt/mTOR信号通路而抑制颗粒细胞自噬，促进卵泡发育成熟，发挥治疗PCOS-IR的作用。最终深入揭示经方桂枝茯苓丸治疗PCOS-IR排卵障碍的作用机制，为该方治疗PCOS-IR提供理论基础，促进其临床应用，提高PCOS临床疗效。

多囊卵巢综合征（PCOS）是一种临床常见的生殖内分泌疾病之一，以持续不排卵、高雄激素或胰岛素抵抗为特征，其在育龄期女性中发病率较高，需终生治疗，并在不同的年龄阶段给女性的身体及心理带来严重影响。胰岛素抵抗在PCOS中起着重要作用，不仅增加了糖尿病等代谢疾病的发病风险，亦是导致排卵障碍的关键因素。本研究基于中医“窠囊”理论，从PCOS的临床表现以及卵巢形态学改变认识到该病的病机为“痰瘀互结，滞碍冲任”，痰湿内盛，邪与血结，阻塞气机，冲任失司，脾失健运，痰瘀内阻，窠囊滞于冲任，壅于胞宫，胞脉，以致难以受孕。根据此理论，临床上采用祛痰利水、活血化瘀的经方桂枝茯苓丸治疗PCOS，并结合当前研究热点肠道菌群，提出“桂枝茯苓丸通过调节肠道菌群改善PCOS的胰岛素抵抗，并通过激活PI3K/Akt/mTOR信号通路调控颗粒细胞自噬，促进卵泡发育成熟，治疗PCOS-IR排卵障碍”的假说。基于此假说，我们建立了PCOS-IR大鼠模型，通过桂枝茯苓丸对模型大鼠进行干预，研究桂枝茯苓丸对模型大鼠肠道菌群、PI3K/Akt/mTOR信号通路及其相关因素的调节作用，初步揭示了桂枝茯苓丸治疗PCOS的作用机制可能是通过调节PCOS-IR大鼠的肠道菌群，改善机体的炎症状态，并通过激活PCOS-IR大鼠卵巢颗粒细胞的PI3K/Akt/mTOR信号通路抑制卵巢的过度自噬，从而促进卵泡的发育。我们还通过直接收集PCOS小鼠的颗粒细胞，采用桂枝茯苓丸含药血清直接培养颗粒细胞，从细胞层面分析桂枝茯苓丸对颗粒细胞PI3K/Akt/mTOR信号通路相关分子表达及自噬的影响，发现桂枝茯苓丸可能通过直接激活颗粒细胞PI3K/Akt/mTOR信号通路，抑制颗粒细胞自噬小体的产生，降低颗粒细胞的自噬率。至此，我们课题组通过在体动物实验和离体细胞实验综合分析了桂枝茯苓丸治疗PCOS排卵障碍的作用机理，为其临床应用提供了现代科学的试验依据。