益肾清利活血方肾靶向调控microRNA表达谱拮抗肾纤维化的机理研究

Renal fibrosis is the common final pathway in chronic kidney disease(CKD), which is regulated by multiple factors. It has been reported that microRNA is involved in the regulation of renal fibrosis recently. Since there is no effective therapy in retarding renal fibrosis, Chinese herbal compound is thought to be the potent therapeutic strategy because of the multi-targeted effectiveness. In our previous multi-center clinical trial, we demonstrated that tonifying kidney clearing and activating blood compound (TCAC) could improve the renal function in patients with CKD stage 3. Furthermore, our basic research studies showed that the main components of TCAC could retard renal fibrosis. In this study, TGF-β1-induced tubular epithelial cell injury in vitro and renal fibrosis rat model in vivo will be established. By using kidney targeting liposome delivery system (KTLDS), we will investigate the role of TCAC-KTLDS on microRNA expression profile and downstream targeting renal fibrosis-related signaling transduction. Moreover, patients with renal failure, characterized by renal fibrosis, will be enrolled and the urine sample will be analyzed to clarify the characters of microRNA expression profile and identify the specific microRNA.

肾纤维化是多种慢性肾脏病(CKD)进展的最终共同通路，受多环节的调控，近年来研究发现microRNA参与了肾纤维化的整个调控过程。肾纤维化目前尚缺乏有效的治疗，中药复方的多组分治疗作用，将是多靶点抗肾纤维化的有效措施之一。我们前期完成的多中心的临床研究提示经验方益肾清利活血方对于CKD3期的患者具有良好的治疗作用，课题组前期分别对该方的有效组分作了相应的基础研究，发现均具有良好的抗肾纤维化作用。本项目将以microRNA为靶点，采用肾靶向复方脂质体技术，利用TGF-β1诱导的肾小管上皮细胞转分化模型及肾纤维化动物模型，研究益肾清利活血组方肾靶向干预，对肾纤维化相关microRNA谱及特异microRNA下游相关肾纤维化信号通路的影响，阐明其通过调控microRNA拮抗肾纤维化的机理。同时筛查慢性肾功能衰竭患者肾纤维化的尿液标本miRNA谱变化，筛查临床特异microRNA。

肾纤维化是多种慢性肾脏病(CKD)进展的最终共同通路，受多环节的调控，近年来研究发现microRNA肾纤维化的调控过程中发挥重要作用。由于肾纤维化目前尚缺乏有效的治疗，中药复方的多组分治疗作用，将是多靶点抗肾纤维化的有效措施之一。基于课题组前期益肾清利活血方对于CKD3期的患者疗效的多中心的临床研究，我们进一步对该方的有效组分作了提取和基础研究，同时采用肾靶向复方脂质体技术，成功建立肾靶向超分子递药系统。在靶向中药复方上，我们成功制作了小复方四味单药主要单体成分的脂质体包裹以及靶向肾脏，同时证实可以在肾脏释放和蓄积。我们构建了TGF-β1诱导的NRK-52E肾小管上皮细胞纤维化模型，并探索中药单体及复方脂质体对TGF-β1诱导的肾小管上皮细胞EMT模型的作用及机制。在研究过程中我们发现衰老与纤维化之间具有相关性，围绕此进一步探究肾脏纤维化与衰老之间共性通路和拮抗相关性（先后性），并进行了清利药黄蜀葵花的主要成分槲皮素、冬虫夏草菌丝在衰老和纤维化方面的相关实验。我们还完成了益肾清利活血组方的网络药理研究及部分验证，对幼龄及老龄鼠进行全转录组测序，筛选了差异microRNA的表达和部分验证。为评估microRNA作为肾脏纤维化生物标志物的临床价值，为进一步探索抗衰老、抗纤维化从而达到肾脏保护作用的研究奠定基础。