基于P38-NF-κB-MCP-1路径的消石利尿化瘀法保护高尿酸血症大鼠血管内皮功能的研究

For uric acid effecting the vascular endothelium function, hyperuricemia can cause serious and extensive harm to body. Eliminating calculus is capable of promoting uric acid metabolism and inhibiting urate crystallization. Diuresis can promote excretion of the uric acid. Removing blood stasis and opening the meridians can protect organism vascular endothelium function. Thus, the hyperuricemia can be treated by using methods of eliminating calculus, diuresis and removing blood stasis. This methods are of innovation and many advantages, for example treating both the root cause and symptoms of disease, and having exact curative effect..Based on the previous research works, this study intended to explore the mechanism of vascular endothelium function to physical injury causing by uric acid in hyperuricemia model. We use methods of eliminating calculus to study the action of lowering uric acid in the experimental rats, and compare them with methods of eliminating calculus and diuresis, removing blood stasis or expelling dampness heat. In this reseach, we test the vasorelaxation factors in oder to discuss the protection of vascular endothelium from serum via morphology of aorta and renal artery, aortic tension. In cytological experiment, we attempt to observe the effects of different medicinal serum to human umbilical vein endothelial cell apoptosis dealed with uric acid. So this project attempts to search the changes of related targets, from p38-NF-κB-COX-2- MCP-1 paths using the models of vascular tissue and cell culture in vitro..Thus, we want to seek out availability and superiority of the method, eliminating calculus, diuresis and removing blood stasis to cure hyperuricemia. Generally speaking, we aim to discuss the mechanism that vascular endothelium function can be protected by eliminating calculus, diuresis and removing blood stasis. The study is supposed to lay the foundation of elevating theoretical level and clinical curative effect of TCM treating hyperuricemia.

高尿酸血症因尿酸损伤血管内皮功能，进而引爆对机体广泛而严重的损害。"消石"促进尿酸代谢，抑制尿酸盐结晶；"利尿"促进尿酸排泄；"化瘀通脉"保护机体血管内皮功能。本课题创新性地以消石利尿化瘀立法组方，兼治标本，疗效确切。.在前期研究基础上，针对尿酸损害机体的关键环节- - 血管内皮功能展开研究，建立高尿酸血症动物模型，与消石利尿法、活血化瘀法及清利湿热法作对比，研究降尿酸作用，通过主动脉、肾动脉形态及主动脉张力研究血管内皮功能保护作用，检测血液中血管舒缩因子，进一步结合含药血清对经尿酸处理后的人脐静脉内皮细胞凋亡的影响，首次通过探查模型血管组织和体外培养细胞因尿酸导致的p38-NF-κB-COX-2- MCP-1路径相关靶点的变化，揭示消石利尿化瘀法治疗高尿酸血症的有效性和优势所在，阐明其保护血管内皮功能的机制，为提升中医药治疗高尿酸血症的理论水平和临床疗效提供实验依据。

高尿酸血症发病率在逐年升高，并基于血管内皮功能的损伤而广泛损害机体。基于尿酸钠盐的结石属性，消石可促进尿酸代谢；由于90%的高尿酸血症是由尿酸排泄减少所引起，利尿可促进尿酸排泄；由于尿酸升高伴发血管内皮功能损伤，化瘀通脉可保护机体，因此，课题组提出以消石利尿化瘀法，针对尿酸损害机体的关键环节——血管内皮功能展开研究。本研究在前期消石利尿化瘀治法降低血清尿酸机理研究的基础上，采用建立高尿酸血症动物模型和含药血清培养经尿酸处理的人脐静脉内皮细胞模型，进行研究。结果表明：以消石利尿化瘀法指导创制的利湿活血方能够显著降低高尿酸血症大鼠模型血清UA含量；可降低高尿酸血症大鼠模型血清PRA，AngⅡ、TXB2、ET-1含量；可升高高尿酸血症大鼠模型血清NO、6-keto-PGF1α含量；可抑制大鼠肾脏ET-1mRNA、COX-2mRNA、MCP-lmRNA表达；提升肾脏组织eNOSmRNA的表达。可减轻主动脉内皮细胞肿胀、脱落及中膜平滑肌细胞排列的紊乱，有一定抑制血管胶原纤维作用；可保护高尿酸血症模型大鼠的肾脏形态。利湿活血方含药血清浓度可上调经高尿酸处理24h、48h、72h后HUVEC活性；含药血清可下调经尿酸处理的人脐静脉内皮细胞p38mRNA、NF-κB mRNA、COX-2mRNA、MCP-lmRNA、ET-1mRNA的表达； 上调eNOSmRNA的表达。通过不同治法（拆方）的对比研究表明：利湿活血方作用与苯溴马隆疗效相当，与四妙丸相比具有显著优势，以清热利湿为主的拆方I与以活血化瘀为主的拆方II比较，I效果优于II，提示清热利湿法是治疗高尿酸血症的关键。以消石利尿化瘀法指导创制的利湿活血方既能有效降低血清尿酸，又能保护肾组织和肾功能，形成良性循环，还能保护血管内皮功能，其作用机制与MAPK应激信号通路有关。将高尿酸血症的防治重心前移，标本兼顾，体现出一定的优势。