从二仙汤配伍研究补肾法对乳腺增生症雌孕激素受体通路的调节

This project is based on a completed funding. We select erxian decoction which has good clinical effect and experimental basis to treat mammary gland hyperplasia as kidney invigorating method representative. The research including in vitro and in vivo experiments is to explore erxian decoction and its decomposed part effect and mechanism. The Kidney Yang invigorating decoction includes Curculigo, Epimedium , Morinda and Angelica, Kidney Yin nourishing decoction includes Anemarrhena, Cork. The effect is studied from breast size, thickness, mammary epithelial cell proliferation by different kidney invigorating method. The mechanism is studied from changes in hormone levels, hormone receptor modulators, receptor response elements, coactivators and coinhibitors. This research includes tissue level, cellular level and molecular level. The purpose is to reveal the mechanism of kidney invigorating method on endocrine disorder of mammary gland hyperplasia.It can offer guideline to the clinic. To investigate the mechanism of erxian decoction can enrich decoction components theory.

本课题在已结题课题基础上，选用临床疗效确切且具有前期实验基础的二仙汤作为补肾治疗乳腺增生症的代表方，以方代法，拟从体内、体外实验两部分探讨二仙汤全方及其拆方：补肾阳方（仙茅、仙灵脾、巴戟天、当归）、滋肾阴方（知母、黄柏）的治疗作用及机制。从乳腺体积、腺体厚度、乳腺上皮细胞增殖情况分别观察二仙汤及其拆方的治疗效应，从激素水平变化、激素受体调节、受体反应元件、辅激活因子、辅抑制因子的变化切入，观察补肾法及其拆方对雌、孕激素受体信号传导通路的调节作用。通过整体、组织水平、细胞水平、分子水平等多层次的研究，探讨二仙汤的组方机理，揭示补肾法对乳腺增生症内分泌紊乱的调节机制，以期为从补肾法论治乳腺增生症奠定实验基础,丰富方剂组方理论的科学内涵。

目的：本课题选用二仙汤作为补肾法治疗乳腺增生症的代表方，拟从体内、体外实验两部分探讨二仙汤全方及其拆方：补肾阳方（仙茅、仙灵脾、巴戟天、当归）、补肾阴泻相火方（知母、黄柏）的治疗作用及机制。结果1.体内实验：与模型组相比，各治疗组乳腺高度均显著下降(P＜0.01)；二仙汤全方中剂量组、补肾阳中剂量组乳腺小叶及腺泡数量减少，分泌现象减弱，导管上皮复层缓解；滋肾阴中剂量组对乳腺病理的改善不明显。WB结果显示：与模型组相比，各治疗组乳腺组织ERα及 SRC-1表达均减少。2.体外实验 各含药组血清组均能降低MCF-10A细胞活力，其中补肾阳拆方中剂量血清与阳性对照组的影响相当。WB结果显示，各含药组血清对MCF-10A细胞的ERα、ERβ、PR、辅激活因子SRC-1及P300的蛋白表达水平均显著下降，辅抑制因子NcoR、SMRT的蛋白表达水平均显著上升。其中补肾阳拆方组与西药组效果较为接近。结论：二仙汤及补肾阳拆方治疗乳腺增生症大鼠的作用机制在于下调乳腺组织中ERα、PR、SRC-1、CBP/P300表达，上调ERβ、辅抑制因子NcoR、SMRT蛋白表达。从而改善大鼠乳腺组织内雌激素受体敏化状态及其受体信号传导通路的异常，对乳腺组织起保护作用。补肾阳拆方在改善雌、孕激素受体敏化及受体信号传导通路异常方面展现了良好的研究前景。