纳米细菌通过NF-κB上调MCP-1导致Ⅲ型前列腺炎发病的机制研究

Category III prostatitis(CP/CPPS) is the most common type of chronic prostatitis. The etiology of CP/CPPS remains unclear and, thus, it is difficult to treat the disease with conventional therapy.Our previous study suggested that Nanobacteria, a newly discovered bacteria, is an important etiological factor for CP/CPPS.It has been showed by our early experiments that MCP-1 is a NB infection related protein which existed in EPS of Patients with CP/CPPS but not in EPS of Patients with another type of prostatitis or without prostatitis. We also found that NF-κB activity and MCP-1 expression were increased significantly by NB infection in prostatic stromal cells, which suggested NF-κB and MCP-1 may paly a role in the pathogenesis of CP/CPPS, but the mechanism remains lagerly unexplored..Based on our data and recent related research progress, we propose a hypothesis that CP/CPPS induced by NB may be mediated by NF-κB activation and the activation related increased expression of MCP-1.In the present study, luciferase reporter assay, EMSA,ChIP,RNAi and confocal microscopy technique were used to test our hypothesis in vitro and in vivo to find a new approach for diagnosis and treatment of CP/CPPS.

Ⅲ型前列腺炎在临床极为常见，但病因有待深入研究。我们率先发现纳米细菌(NB)是Ⅲ型前列腺炎重要病因，但其机制未见报道。前期实验证实，单核细胞趋化蛋白MCP-1是NB引起的Ⅲ型前列腺炎与其他前列腺炎及正常人前列腺液中的主要差异蛋白；进而用NB攻击前列腺基质细胞，其转录因子NF-κB活性增强、MCP-1表达增加，但NB致MCP-1上调及引发前列腺炎的具体机制尚不清。我们推测NB可能通过活化NF-κB信号转导通路上调MCP-1继而导致前列腺炎。为证实此推测，本项目拟借助荧光素酶报告基因检测、EMSA、ChIP、RNA干扰及激光共聚焦等技术，解析NB通过活化NF-κB进而在转录水平上调MCP-1表达的机制；并在动物模型中证实NB上调MCP-1及导致Ⅲ型前列腺炎发生的机制。本项目旨在揭示NB上调MCP-1的分子机制及其在Ⅲ型前列腺炎发病中的意义，为其诊治提供依据。

Ⅲ型前列腺炎在临床极为常见，但病因有待深入研究。我们率先发现纳米细菌(NB)是Ⅲ型前列腺炎重要病因，但其机制未见报道。前期实验证实，单核细胞趋化蛋白MCP-1是NB引起的Ⅲ型前列腺炎与其他前列腺炎及正常人前列腺液中的主要差异蛋白；进而用NB攻击前列腺基质细胞，其转录因子NF-κB活性增强、MCP-1表达增加，但NB致MCP-1上调及引发前列腺炎的具体机制尚不清。我们推测NB可能通过活化NF-κB信号转导通路上调MCP-1继而导致前列腺炎。为证实此推测，我们将收集的NB致CP/CPPS患者EPS蛋白质、非NB致CP/CPPS患者EPS蛋白质及正常人EPS蛋白质，进行双向电泳和质谱分析后，发现NB致CP/CPPS患者前列腺液中的MCP-1含量升高，是主要差异性蛋白之一。NB作用于前列腺基质细胞WPMY-1后，激光共聚焦显微镜提示，细胞内转录因子NF-κB活性增强。MCP-1 基因启动子中含有NF-κB结合位点，继而我们成功获得MCP-1基因启动子区域，经荧光素酶(Luc)报告基因检测证实，NB感染WPMY-1细胞后，MCP-1基因启动子区域活性增强。实验结果显示， CP/CPPS患者前列腺液标本中存在NB，并且有较高的阳性率。NB作用于前列腺基质细胞（WPMY-1）后，细胞中转录因子NF-κB激活，进而与MCP-1 5’端启动子区（-670~ -662）的位点结合，增强MCP-1基因启动子活性，从而证实了NB在转录水平上促使MCP-1表达的具体分子机制，为NB致CP/CPPS发病的机制提供了一些理论依据。在本研究中我们还针对性激素受体包括雄激素受体(AR)、雌激素受体α(ERα)、雌激素受体β(ERβ)和孕激素受体PGR)在BPH中的可能机制做一探索性研究，期望能找到一个突破口。在人BPH组织中AR和PGR表达显著升高，ERα的表达显著下降；大鼠BPH模型实验组中PGR的表达显著上调，AR和ERα的表达显著下调；ERβ的表达未见显著性差异；IHC、qRT-PCR、WB在检测上述性激素受体表达的结果一致。