基于NF-κB通路的蒙药玉簪花抗Ⅲ型前列腺炎活性成分及作用机制研究

The flowers of Hosta plantaginea (Lam.) Aschers (HPF) has a long history and commonly used Mongolian traditional medicine with specific curative effect for the treatment of sore throats, hoarseness, pulmonary heat and toxic heat. Through private investigation it was found that HPF has good effect for the treatment of chronic prostatitis, but its material bases and mechanisms are unknown. In our pre-experiment, we had proved that ethyl acetate and n-butanol solvent extracts from HPF showed strong protective effect as the positive control Qianliekang against Xiaozhiling-induced experimental CP/CPPS model in rats. It was also indicated that the two active extracts have anti-CP/CPPS properties with decreasing expression levels of inflammatory cytokines, such as TNF-α, IL-1β, and IL-8. In order to investigate the material bases and mechanisms of HPF for anti-CP/CPPS, the chemical constituents will be separated, purified, and identified from the two active extracts. All compounds will be screened for anti-inflammatory activities, and to investigate the expression change of relevant inflammatory cytokines, enzymes, and proteins in the models of cellular and molecular. Further mechanisms of the bioactive compounds will be studied base on the inflammation related NF-κB signaling pathway. Confirm the therapeutic effect of active compounds in CP/CPPS rat model. All this will elucidate the scientific connotation of Mongolian medicine HPF in treating CP/CPPS disease. We hope that the results of the study can provide theoretical basis for the utilization of HPF, and to provide promising leading compounds for the development of natural anti-CP/CPPS medicine.

玉簪花是具有悠久蒙医临床用药历史和确切疗效的常用药材，用于治疗咽喉肿痛、喑哑、肺热和毒热，民间也用于治疗慢性前列腺炎，但其抗炎的物质基础及作用机制不明确。前期实验证实，玉簪花乙酸乙酯和正丁醇部位对Ⅲ型前列腺炎模型大鼠具有很好的治疗作用，其活性优于阳性对照前列康，并提示其可能与调节炎症因子水平（TNF-α、IL-1β、IL-8等）有关。为了深入研究玉簪花抗Ⅲ型前列腺炎作用的药效物质基础及作用机制，拟采用Ⅲ型前列腺炎整体动物模型活性追踪为导向进行有效部位的分离纯化及结构鉴定；测定单体化合物在细胞水平和分子水平上对多种相关炎症因子、酶活性及蛋白表达的含量变化，并探讨对NF-κB信号通路的影响；然后通过Ⅲ型前列腺炎动物模型进一步证实活性化合物的治疗作用。为蒙药玉簪花的临床用药和合理开发提供科学依据，也为天然抗Ⅲ型前列腺炎的研发提供先导化合物。

玉簪花是蒙医传统常用药材，用于治疗咽喉肿痛、肺热等，民间也用于治疗慢性前列腺炎，但其化学成分和药理活性研究较薄弱、药效物质基础及作用机制尚不明确。本课题对玉簪花进行了系统的化学成分、活性评价、总黄酮提取工艺等研究，主要结果如下：1. 80%乙醇提取物在低、中、高剂量下对二甲苯致小鼠耳廓肿胀均表现出较强的抑制作用，优于其它乙醇体积分数提取物，并且能更好的富集色谱峰。2.乙酸乙酯（B）和正丁醇部位（C）对消痔灵所致CP/CPPS大鼠模型具有较好的治疗作用，为有效部位。3.从有效部位中分离并鉴定了41个单体化合物，其中新化合物6个、新天然产物1个、属首分13个、种首分4个，为首次从百合科中分离得到二氢黄酮类和β-卡巴林酸类化合物。4.黄酮类化合物具有较强的抑制环氧化酶（COX）1和2活性、NO抑制作用及较强的抗氧化活性，并显示出一定的构-效（抗氧化）关系，提示黄酮类成分可能通过抗炎和抗氧化来治疗CP/CPPS，与前期文献报道一致。5.玉簪花各萃取部位总黄酮含量B > A（石油醚部位）> C > D（水部位）、总多酚含量B > C > A > D，DPPH抑制作用B > A > D > C、ABTS抑制作用C ≈ B > A > D。此外，总提取物和B具有很强的α-葡萄糖苷酶抑制活性，强于阳性药阿卡波糖。6.对我国四种玉簪属植物玉簪（HP）、紫萼（HV）、东北玉簪（HE）和紫玉簪（HA）进行了总黄酮、总多酚及其抗氧化活性和α-葡萄糖苷酶活性比较研究，结果表明，HA的总黄酮和总多酚含量高，且具有很强的α-葡萄糖苷酶抑制活性。7.采用单因素试验和响应面法建立了玉簪花总黄酮的最佳提取工艺。通过本项目的研究，基本阐明了玉簪花抗CP/CPPS的物质基础及作用机制，为其临床合理用药和综合开发提供了科学依据，也为天然抗CP/CPPS的药物研发提供参考，完成了课题预期目标。