广西特产黑蚁活性组分介于改善高尿酸血症治疗痛风的作用机制研究

Hyperuricemia belongs to the category of "Bi-syndrome" in traditional Chinese medicine. Hyperuricemia is caused by the disorder of purin metabo1ism and is an important pathological basis for gout, furthermore is closely associated with some serious diseases such as hypertension. At present, the antihyperuricemic agents on the market possess some side effects which hinder their application in clinical practice. Therefore, searches for alternative antihyperuricemic agents with a more favorable pharmacological and toxicological profile is highly warranted. Polyrhachis vicina Roger，a well-known traditional Chinese medicine，are quite rich in Guangxi,but R&D about it have been lagging behind. With the clues from some Chinese medical books and its long time application in clinical practice in China, we verified, through years of researches with the help of the modern science technologies, that Polyrhachisvicina Roger can alleviate gout by decreasing the level of uric acid in serum(by inhibiting the activity of XOD) and increasing the level of uric acid in urine. But the underlying mechnism is not yet elucidated. To explore the mechanism of the anti-hyperuricemia activity of these active constituents extracte from Polyrhachis vicina Roger, with the guidance of the traditional Chinese medicine theory, the project will be carried out to study the effects of Polyrhachisvicina Roger on the inhibition of the uric acid overproduction and underexcretion in the hyperuricemia model rats and the Clone-9 cells at the different levels of the whole body, organs, tissues and cells by RT-PCR, immunohistochemistry and Western blot, subsequently laying the foundation for developmenting Polyrhachisvicina Roger as the antihyperuricemic agents with the independent intellectual properties.

高尿酸血症归属中医"痹证"。高尿酸血症由嘌呤代谢紊乱所致，是引起痛风的生化基础，且与高血压等疾病密切相关，目前治疗药物毒副作用大而限制其在临床上的使用，因此，研制高效低毒的抗高尿酸血症药物具有重要的意义。 广西特产黑蚁为广西道地药材，资源丰富，但研究与开发程度低。本课题组以中医药典籍记载及民间临床实践为线索，经过多年研究证实了其抗痛风活性，并初步揭示其抗痛风作用是通过减少血清尿酸水平（抑制XOD的活性）和增加尿中尿酸水平密切相关,但其作用机制尚需系统解明。为系统阐明其抗高尿酸血症的作用机制，本项目拟以中医药理论为指导，通过建立高尿酸血症大鼠模型和培育大鼠Clone-9细胞，利用RT-PCR、免疫组化和Western-blot等技术，从整体、器官、细胞和分子水平多层次对其活性组分干预尿酸的合成体系及尿酸在肾脏排泄体系两方面展开系统研究，为研制具自主知识产权的抗高尿酸血症药物奠定基础。

本项目采用氧嗪酸钾诱导大鼠，成功建立大鼠高尿酸血症模型。口服给予广西特产黑蚁活性组分（AFPR），检测大鼠尿液和血清中的尿酸水平，证实AFPR对高尿酸血症大鼠具有治疗作用；测定AFPR对高尿酸血症大鼠血清和肝脏中黄嘌呤氧化酶（XOD）水平，血清SOD活性及IL-1β、IL-6、 TNF-α、SCr、BUN和MDA的水平，肾脏尿酸盐转运蛋白（GLUT9，URAT1，OAT1）的表达，证明AFPR具有抗氧化、抗炎活性和对肾具有保护作用，部分阐明AFPR治疗高尿酸血症相关分子机制；通过尿酸处理大鼠肝细胞诱发炎症，建立尿酸诱导大鼠肝细胞炎症模型，给予AFPR处理，检测细胞炎症因子IL-1β、IL-6和TNF-α的表达水平，证实AFPR能有效抑制高尿酸诱发的炎症及其机制，进一步阐明广西特产黑蚁活性组分抗高尿酸血症作用的多靶点性；使用氢氧化钾甲醇溶液将广西特产黑蚁中的脂肪酸甲酯化，采用正十八烷作为内标物，建立拟黑多刺蚁中棕榈酸和油酸含量的气相色谱测定方法。本研究为研制具自主知识产权的抗高尿酸血症药物奠定基础。. 在本项目资助下，发表论文7篇，其中SCI论文1篇，中文核心期刊6篇。获得国家发明专利授权2项。项目总投入经费49.00万元，支出49.00万元，各项支出基本与预算相符。