基于雌/雄激素调控网络的补骨脂主要活性成分抑制良性前列腺增生的分子机制研究

Benign prostatic hyperplasia (BPH) is a common disease with high incidence in older men. It is well known that the enhanced effect of estrogen/androgen reaction and the imbalance of their ratio play important roles during the BPH process and the main treatment for BPH is medication. Psoralen corylifolia is one of the tradiitional Chinese medicine which is most widely used for BPH. We previously found that its components of Coumarin（including psoralen and isopsoralen）and Monoterpnoid (bakuchiol) are the main active compounents that inhibited the prostatic cell proliferation，and furtherfore， different components aboved have also showed their efficacy on estrogen receptor (ER) activation, androgen receptor (AR) antagonism and expression inhibition of aromatase which is the key enzyme converting androgen to estrogen.Our study will focus on the active components mentioned above and on the basis of the results we got, we will further do the research on their mechanism for BPH development by the multi-biochemical methods and try to give an illustrate that whether ER、AR and aromatase are involved. In prostatic stromal and epithelial cells, the expressions, locations and promoter activities of the different targets including ER, AR and aromatase will be determined for the active components. The effects of prostatic stromal and epithelial cell proliferation, phenotype transformation from fibroblasts to smooth muscle cells, epithelial to mesenchymal transition (EMT) and stromal-epithelial interactions will then be identified. Using the ChIP-seq technique the signal pathways involved will be analyzed. Using ER, AR or aromatase knock-out mice, we will further confirm the mechanism of Psoralen in the development of BPH and the role of component- related-targets involved. and draw a map for "drug - target - pathway - disease". The results will provide the pharmacologic evidence and pathways for the integral treatment of Psoralen with multi-component and multi-target in BPH with regard to androgen and estrogen regulation network, and will also provide to develop inovative, safe and effective traditional Chinese medicine for BPH treatment.

良性前列腺增生（BPH）是高龄男性的常见病多发病。雌/雄激素效应增强和比例失调是主要诱因，药物是主要治疗手段。补骨脂是最常用的治疗BPH的中药，前期研究发现其香豆素类成分（补骨脂素、异补骨脂素）和单萜类成分（补骨脂酚）是抑制前列腺细胞增殖的主要活性成分，并且不同成分还兼有激活雌激素受体（ER）、拮抗雄激素受体（AR）和抑制雌/雄激素转化酶芳香化酶表达的作用。本项目选择上述两类活性成分在前期研究基础上，采用多种生物技术进一步确认其在前列腺间质和上皮细胞中对ER、AR和芳香化酶的表达、定位和启动子活性的调控作用，阐述其对前列腺间质和上皮细胞增殖、间质平滑肌表型、上皮间质化和间质-上皮相互作用的影响；初步解析通路；用靶标敲除的动物模型进行验证；实现"药物-靶标-通路-疾病"的对接。从雌/雄激素网络调控角度为补骨脂多成分多靶点协同治疗BPH提供药理学依据，也为开发创新型BPH安全有效药物提供思路。

良性前列腺增生症（Benign Prostatic Hyperplasia，BPH） 是老年男性泌尿系统的常见病，高发病。临床上采用以抗雄／抗雌药物抑制前列腺体积的增大治疗BPH。本项目选取BPH治疗时常用中药补骨脂三个主要成分为研究对象围绕雌雄激素信号通路阐述其抗BPH的物质基础和作用机制。补骨脂酚（ba）补骨脂素（pso）和异补骨脂素（iso）均明显抑制前列腺间质细胞WPMY-1和上皮细胞BPH-1的增殖；同时促进BPH-1细胞中抗增殖作用的ERβ表达，对BPH-1细胞中AR和WPMY-1细胞中ERα作用不明显；ba而不是pso或iso可明显抑制WPMY-1细胞中芳香化酶的表达。ba通过ERβ 而非Erα特异性地拮抗BPH-1细胞中雌二醇诱导的雌激素应答元件ERE的活性。另外，ba抑制芳香化酶的表达与PGE2信号通路相关。在体实验证实，ba可以抑制雌／雄激素诱导的大鼠BPH进程，这与抑制芳香化酶表达，促进ERβ表达密切相关。下尿路状况（lower urinary tract symptoms，LUTs)包括指膀胱逼尿肌的过度收缩和排尿功能异常等症状，在BPH患者中有极高的发病率。传统中药黄连(Coptis chiensis Franch, CSF)用于肠道具有抗平滑肌收缩的作用。本项目还研究了黄连提取物抗BPH相关的LUTs的作用。发现和BPH模型组相比，CSF可以明显抑制BPH大鼠的前列腺膀胱的指数和病理改变；排尿间隔和排尿收缩时间明显延长，排尿体积明显增加，排尿时间明显缩短。器官水平上，CSF明显抑制KCl或乙酰胆碱诱发的逼尿肌的过度收缩，改善大鼠膀胱排尿参数，与在体结果一致。本项目从雌／雄激素网络调控角度为补骨脂抗BPH治疗提供了药理学依据，从改善膀胱下尿路状况角度为开发黄连提取物用于BPH相关LUTs的创新型BPH安全有效药物提供了思路。