基于TGF-β/Smad与Wnt/β-catenin通路研究经方桃核承气汤干预慢性肾衰竭大鼠肾纤维化

TGF-β/Smad and Wnt/β-catenin signaling pathway were both closely related to renal fibrosis. But it is still unclea what is their crosslinked relationship in regulating renal fibrosis of chronic renal failure(CRF).It was thought that toxin and blood stasis were major cause of CRF in traditional Chinese medicine. Classic prescription THCQT could expel stasis and attack toxin. Our previous study demonstrated that this prescription could delay the process of CRF and improve renal fibrosis. However the exact mechanism is not clear. This project will base on the 5/6 renal removal of the rat model of CRF as a carrier, use Western blot and Quantitative Real-time PCR technology to detect associated factor protein content and mRNA expression of TGF-β/Smad and Wnt/β-catenin signaling pathway in kidney tissue, from the level of modern molecular biology and morphology. This project will explore molecular mechanisms of renal fibrosis in CRF rat intervened by THCQT. Therefore it will provide the scientific basis for classic prescription treating CRF, and lay a foundation for its clinical application. Meanwhile, establishing the initial relationship between two paths and their crosslinked ,which will play a role in renal fibrosis of CRF and its curing, will provide reference for clinical screening of drugs.

TGF-β/Smad与Wnt/β-catenin信号通路均和肾纤维化进程密切相关，但此两条通路在调控慢性肾衰竭（CRF）肾纤维化过程中的交联关系尚不明确。中医认为毒邪与瘀血是CRF的主要病因，经方桃核承气汤具有破血化瘀、攻逐毒邪之功效。本课题组前期研究证实该方能延缓CRF进程，改善肾纤维化，但具体机制尚不明确。本项目以5/6肾切除CRF大鼠模型为载体，从现代分子生物学和形态学层面，分别采用Western blot和实时荧光定量PCR技术检测肾组织中与TGF-β/Smad及Wnt/β-catenin信号通路相关因子的蛋白含量及其mRNA的表达，探讨桃核承气汤干预CRF大鼠肾纤维化的分子机制，为经方治疗CRF提供科学依据，为其临床推广应用奠定基础。同时，初步确立此二条通路及其交联关系在CRF肾纤维化发生及治疗中的作用，为临床有效治疗药物的筛选提供借鉴。

TGF-β/Smad与Wnt/β-catenin信号通路均和肾纤维化进程密切相关，但此两条通路在调控慢性肾衰竭（CRF）肾纤维化过程中的交联关系尚不明确。中医认为毒邪与瘀血是CRF的主要病因，经方桃核承气汤具有破血化瘀、攻逐毒邪之功效。本课题组前期研究证实该方能延缓CRF进程，改善肾纤维化，但具体机制尚不明确。本项目以5/6肾切除CRF大鼠模型为载体，从现代分子生物学和形态学层面，分别采用Western blot和实时荧光定量PCR技术检测肾组织中与TGF-β/Smad及Wnt/β-catenin信号通路相关因子的蛋白含量及其mRNA的表达，探讨桃核承气汤干预CRF大鼠肾纤维化的分子机制。发现桃核承气汤对CRF大鼠Wnt/β-catenin信号通路的影响：抑制Wnt1、Wnt3a、Wnt4、Wnt5a、p-GSK-3β、β-catenin等蛋白，促进Wnt5b、E-cadherin、Axin等蛋白；对TGF-β1/Smad3信号通路的影响：抑制TGF-β1、Smad3、CTGF等蛋白，促进Smad7、miR-29a、miR-29b、miR-29c等蛋白。这些研究成果为经方治疗CRF提供科学依据，为其临床推广应用奠定基础。同时，初步确立此二条通路及其交联关系在CRF肾纤维化发生及治疗中的作用，为临床有效治疗药物的筛选提供借鉴。