从miRNA调控caspase凋亡通路探讨前列宁胶囊治疗良性前列腺增生的作用机制

Deregulation of miRNA impairs the caspase-mediated apoptosis, which therefore is strongly correlated with the pathogenesis and development of benign prostatic hyperplasia (BPH). 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 BPH. From our previous studies supported by the Nature Science Foundation of China, we found that QC could promote BPH cell apoptosis via regulating the expression of some target gene of caspase apototic pathway. Furthermore, QC could modulate the expression of multiple apoptosis-related miRNAs, suggesting that QC treats BPH probably through regulation of miRNA leading to the caspase activation. 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-mediated caspase apototic pathway and its downstrem target gene expression. 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, providing theoretic foundation for QC's clinical application.

miRNA调控异常导致的caspase凋亡通路受阻是良性前列腺增生（BPH）发病的重要机制。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BPH疗效确切。课题组在前两个国家基金资助下研究发现，前列宁胶囊能够调节caspase内源性凋亡通路相关基因的表达从而诱导BPH细胞凋亡；进一步研究发现前列宁胶囊能够调控多个与凋亡相关的miRNA，提示前列宁胶囊通过调控miRNA而激活caspase凋亡通路可能是其治疗BPH的重要作用机制。本课题在此基础上拟以BPH动物及细胞模型为研究对象，运用分子克隆、细胞转染、real-time PCR、western blot等技术研究前列宁胶囊对miRNA及其调控的caspase凋亡通路和相关靶基因表达的影响，进一步系统而深入地从表观遗传学的角度阐明前列宁胶囊治疗BPH的作用机制，明确前列宁胶囊治疗BPH 的作用靶点，为前列宁胶囊的临床应用奠定理论基础。

本研究结合动物实验和体外实验，采用Real time-PCR、Western-Blot、流式细胞术和细胞转染等方法对前列宁胶囊（Qianliening Capsule，QC) 治疗良性前列腺增生症（Benign Prostatic Hyperplasia，BPH）的促凋亡分子机制展开了深入研究。研究结果发现：⑴ QC能明显改善BPH模型大鼠的前列腺增生情况，提示QC对BPH大鼠有显著的疗效； ⑵ 动物实验发现QC能调节凋亡相关基因Bax、Bcl-2和FAS的表达，促进BPH模型大鼠中前列腺组织细胞的凋亡； ⑶ 体外细胞实验发现QC能够改变BPH-1细胞线粒体膜电位，释放细胞色素C，激活下游Caspase-8和Caspase-9 的活化，进一步活化Caspase-3，提示QC对BPH的治疗作用是激活Caspase途径，从而引起BPH1细胞凋亡的增加；（4）细胞转染结果表明，高表达miR-34C-5P能够诱导BPH1细胞凋亡，进一步明确了QC的作用靶点。上述结果深入地揭示了QC的促凋亡分子机制和作用靶点，为QC的临床应用奠定了实验依据。