基于COX和LOX双靶点调控的滋肾丸治疗前列腺增生的物质基础及作用机制研究

The multi-components and multi-targets approach of TCM has many advantages than single chemical entity in dealing with chronic diseases. Zishen Pill is a classical Chinese herbal formula used for treatment of benign prostatic hyperplasia (BPH). Our previous study showed that the pathological conditions of BPH could be by partially reversed by inhibition of metabolite levels of arachidonic acid(AA) via COX and LOX pathways. Base on this observation, enzyme active ingredients could be screened, the AA dynamic regulatory networks could be re-constructed with different combination of bioactive components of this formula binding to COX or LOX and relative gene Bcl-2. In this proposal, by quantifying the metabolites, gene expression, the underlying mechanisms will be investigated by using metabonomic, molecular biology and bioinformatics. This study will not only provide a targeted molecular description of therapeutic basis of Zishen Pill, but also validate the concept that multi-targeted and multi-components of traditional Chinese medicine would have a better chance for prevention and treatment of chronic diseases, such as BPH.

中药复方具有多成分、多靶点的效应，从而在慢性疾病中更能体现出优于单分子、单靶点药物的疗效优势。经典方滋肾丸对治疗前列腺增生症(BPH)具有确切疗效。前期研究证实该方对BPH炎症花生四烯酸(AA)代谢COX和LOX路径具有双重抑制、多靶点的效应，但具体的作用机制尚不明确。本项目从体外筛选抑制上述酶的活性成分着手，采用“体外活性分子—体内药效成分—体内靶标成分”逐层推进的策略，基于AA/COX,LOX代谢通路及其调控的Bcl-2凋亡基因，构建与不同活性成分(群)及组合特征相对应的AA代谢网络动态调控模型，整合反映网络质与量变化的代谢物、基因、蛋白指标，与体内化学物质组变化相关联，发现靶标成分，阐释其作用靶点和作用机制，深入探讨滋肾丸对疾病靶点生物途径的效应，进而验证组方的协同整体特征。结合代谢组学、分子生物学、生物信息学方法，建立一种基于特定调控靶点捕获、辨识BPH治疗药物有效成分的新途径。

经典方滋肾丸对治疗前列腺增生症(BPH)具有确切疗效。研究证实该方对BPH炎症花生四烯酸(AA)代谢COX和LOX路径具有双重抑制、多靶点的效应。本项目从体外活性成分着手，以COX-2/5-LOX酶为靶标生物大分子，采用亲和超滤HPLC-MS法筛选出方中潜在的COX-2/5-LOX酶抑制剂。选择上述双靶点, 基于靶向代谢组学方法构建与不同活性成分(群)及组合（药对）特征相对应的AA代谢网络动态调控模型，采用UHPLC-MS/MS法对BPH模型大鼠与正常大鼠体内血浆、组织中AA代谢轮廓进行定量分析，整合反映机体网络变化的代谢物、蛋白和基因为效应指标，将方剂化学物质组的变化与生物系统的应答响应相关联，发现了靶标成分，阐释作用靶点，深入探讨了滋肾丸基于AA/COX,LOX代谢通路及其调控的Bcl-2细胞凋亡生物途径的作用机制，进而科学阐释中药组方配伍的合理性。