补肾益精法对系统性硬皮病血管损伤的保护机制：拮抗Snail介导的EndMT所致血管损伤?

补肾益精法是邓铁涛教授辨治硬皮病的经验治法；主张从“肺脾肾相关理论” 辨治硬皮病、“补肾益精为主，健脾养肺为辅”。前期研究确定了抗纤维化的主要机制与有效成分；同时，也发现其对硬皮病血管的保护作用：补肾益精法可增加皮损区毛细血管数量，内皮细胞表达α-SMA减少；在体外可抑制血管内皮细胞-间质转化（EndMT）过程，且伴随核内Snail水平回落。然而，具体机制不明确。因此，结合传统中医肾与血脉相关理论，提出研究假说：补肾益精法对硬皮病血管损伤的保护作用，可能通过拮抗Snail介导的EndMT过程、抑制纤维增生性闭塞性血管损伤而产生？本项目拟在体外与体内两个层面，采用免疫荧光、PCR、Western Blot、质粒过表达、RNA干扰、细胞谱系示踪等技术，阐明补肾益精法对硬皮病血管保护作用的相关机制，为中医治疗疑难免疫性疾病提供科学依据，为硬皮病治疗提供新的干预策略，为传统中医理论夯实科学基础。

Tonifying kidney and Boosting essence treatment is the therapy for systemic sclerosis from Prof. Deng, who recommended treating this disease on the theory of “Lung-Spleen-Kidney Correlation”, and “tonifying kidney and boosting essence at the first place, strengthening spleen and nourishing lung at the supplement place”. In the previous projects on this therapy, we found the anti-fibrotic mechanism and its main gradient; meanwhile, we also discovered its protective role in proliferative vasculopathy of SSc: this therapy increased the number of capillaries in the fibrotic legion, and decreased the amount of α-SMA positive endothelial cells; in TGF-β induced endothelial to mesenchymal transition(EndMT) cell models, this treatment inhibited the transition from endothelial to mesenchymal cells, correlated with declined snail levels in the nuclei. Thus, having referred to the classical theory on the relation between kidney and vessel, we proposed the hypothesis: one of therapeutic mechanisms of Tonifying kidney and Boosting essence treatment on proliferative vasculopathy in systemic sclerosis could be that the Chinese medicine therapy might antagonize vascular injuries caused by the snail mediated EndMT? This project would be carried out in vivo and in vitro, by utilizing techniques such as immunofluorescence, PCR, Western Blot, gene over-expression or interference by RNA interference, cell lineage tracing, etc., to comprehensively study the mechanisms of the Tonifying kidney and Boosting essence treatment on vascular protection in systemic sclerosis, so as to provide substantial evidence for this classical Chinese therapy, to possibly suppose an EndMT-targeted strategy in the treatment of systemic sclerosis, and to strengthen the basis of Chinese medicine theory.