基于microRNA调控网络的前列宁胶囊治疗良性前列腺增生的作用机制研究

Qianliening capsule （QC）is a hospital preparation at the affiliated hospital of Fujian University of Traditional Chinese Medicine. QC has been shown to be effective in the clinical treatment of benign prostatic hyperplasia (BPH). BPH is a complex disease because its pathogenesis and development is associated with multiple factors, genes and signal transduction pathways, all of which are further highly regulated by miRNA regulatory network. Our previous studies suggest that, as a multi-component and multi-target agent, QC may exert its anti-BPH function via affecting the miRNA regulatory network in prostatic cells. To further investigate the pathogenesis of BPH and the precise molecular mechanisms of QC's therapeutic activity, in present project we will set up BPH animal and cell models, analyze the miRNA, mRNA and protein expression profiles in BPH, and then construct the miRNA-based BPH disease-network. Moreover, we will study the effects of QC in vivo and in vitro on the expression of those specifically expressed miRNA, mRNA, protein and relevant signaling pathways in BPH. Furthermore, using bioinformatics technology we will analyze our massive experimental data and construct a 'disease-target-drug' network accordingly. Our findings in this project will provide new insight into the molecular mechanism of pathogenesis of BPH, as well as fully elucidate the mechanism of action of how QC treats BPH.

前列宁胶囊是福建中医药大学附属人民医院院内制剂，治疗良性前列腺增生（BPH）疗效确切。BPH是多因素、多基因、多通路参与调控的复杂性疾病。现代表观遗传学研究表明miRNA调控网络对BPH发病机制相关的多条信号转导通路有重要调节作用。课题组前期研究表明前列宁胶囊可调节BPH的多条信号通路及相关基因的表达，提示多成分、多靶点协同干预miRNA调控网络可能是前列宁胶囊治疗BPH的重要机制。本课题拟应用BPH动物及细胞模型，运用组学技术分析BPH的miRNA、mRNA和蛋白特异性差异表达，构建BPH的miRNA调控网络，以进一步明确BPH发病机制；进而用前列宁胶囊干预，研究其对BPH特异性差异表达的miRNA及其靶基因和相关信号通路的影响，用生物信息学技术对实验数据进行分析，构建疾病-靶点-药物网络，全面阐明前列宁胶囊治疗BPH的作用机制，为前列宁胶囊的临床应用提供理论依据。

本研究通过细胞实验，利用miRNA和mRNA表达谱芯片筛选、realtime PCR验证及其生物信息分析等对前列宁胶囊（Qianliening Capsule，QC) 治疗良性前列腺增生症（Benign Prostatic Hyperplasia，BPH）的分子作用机制进行深入研究。研究结果发现：⑴ QC能明显抑制BPH-1细胞活力，提示QC对BPH-1细胞增殖有明显抑制作用； ⑵ miRNA表达谱芯片检测结果显示：重复样本间相关性良好，与对照组相比，加药组有17个差异miRNA升高（fold change＞2），2个miRNA下降（fold change＜-2），realtime PCR验证结果与芯片结果相符； ⑶ mRNA表达谱芯片检测结果显示：重复样本间相关性良好，与对照组相比，加药组有107个差异mRNA升高（fold change＞2），71个mRNA下降（fold change＜-2），realtime PCR验证结果与芯片结果相符；（4）生物信息学分析显示：GO和PATHWAY分析表明药物的作用主要与细胞的增殖与凋亡关系密切。此外，应用Cytoscape等生物信息学方法对实验所得数据进行整合分析，构建QC治疗BPH的miRNA-mRNA调控网络。上述结果全面阐明前列宁胶囊治疗BPH的作用机制，为前列宁胶囊的临床应用提供理论依据。