miR145介导ERα下调SAA1在III型前列腺炎发生中的作用机制研究

Type III prostatitis(CP/CPPS) is the most common chronic prostatitis. The etiology of CP/CPPS remains unclear. It is reported ERα closely related with CP/CPPS; serum amyloid protein A1 (SAA1) and micro RNA 145 (miR145) contribute to the inflammation as well. In the previous study, microRNA and gene expression chip showed that miR145 was low expression, while ERα and serum amyloid SAA1 were high expression in prostate tissu of CP/CPPS. Report assays demonstrated that activated ERα could down-regulate miR145. And miR145 could down-regulate SAA1. Bioinformatics analysis found there were ERα potential binding site in the promoter of miR145 and miR145 potential binding site in the 3'-UTR of SAA1. However, the mechanism of regulation and the relationship among ERα, miR145 and SAA1 is presently unknown. We supposed that miR145 could suppress the expression of SAA1. Activated ERα could down-regulate miR145 in CP/CPPS. Then SAA1 could up-regulate in CP/CPPS.In this study, we will reveal the mechanism of the pathway of ERα-miR145-SAA1 in CP/CPPS by report assay, EMSA, ChIP and RNAi et al. It may find a new approach for diagnosis and treatment of CP/CPPS.

Ⅲ型前列腺炎(CP/CPPS)是泌尿科最常见疾病，病因不清，而ERα在CP/CPPS起重要作用,SAA1、miR145与炎症关系密切。预实验在microRNA芯片、表达谱芯片结果中发现，CP/CPPS前列腺组织标本中miR145低表达，ERα、SAA1高表达。报告基因实验证实，活化的ERα可降低miR145启动子活性，而miR145可作用于SAA1的3'UTR降低其表达；生物信息学分析均发现潜在结合位点。ERα、miR145、SAA1三者在CP/CPPS中的作用及其分子机制不清。推测miR145可抑制SAA1表达；CP/CPPS发生过程中ERα活化，作用于miR145启动子使其表达减少，进而使SAA1高表达。本项目拟采用Luc报告基因、EMSA、ChIP、RNA干扰等方法，揭示miR145介导ERα上调SAA1的分子机制及其在CP/CPPS发生、发展中的作用，为该病诊治提供新的理论基础。

Ⅲ型前列腺炎 (CP/CPPS)是泌尿外科最常见的疾病之一，由于病因复杂，治疗效果不佳，对其发病机制的研究受到广泛关注。有研究表明雌激素与CP/CPPS的发生、发展密切相关。雌激素在体内可经雌激素受体α (ERα)信号途径作用于前列腺间质引起组织炎症，但其具体分子机制尚不清。在前期实验研究中，microRNA芯片结果显示，CP/CPPS前列腺组织中miR145表达较正常前列腺组织下降。表达谱芯片提示，ERα及炎症相关因子血清淀粉样蛋白A1(SAA1)在CP/CPPS前列腺组织中表达增多。进一步生物信息学分析miR145基因5’调控区有潜在的ERα结合位点，报告基因实验证实，活化的ERα可使miR145基因5’调控区活性降低。SAA1的 3'-UTR有miR145可能作用位点，而miR145也的确可下调SAA1 3'-UTR活性。进而我们在人前列腺基质细胞中明确，ERα活化后可抑制miR145表达，并且导致SAA1高表达；miR145的5＇调控区-359~-350有ERα结合位点，并可下调miR145表达；SAA1的3'-UTR存在miR145结合位点。本项目旨在揭示miR145介导ERα上调SAA1的分子机制及其在CP/CPPS发生、发展中的作用，为该疾病发生机制及临床治疗提供新思路。除此之外，我们还探讨了外泌体在CP/CPPS患者前列腺液中数量明显增多，可能在该疾病发病机制中具有重要作用；同时我们还针对性激素受体包括雄激素受体(AR)、ERα、雌激素受体β(ERβ)和孕激素受体(PGR)在BPH中的可能机制做了探索性研究，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。