动脉粥样硬化(AS)是冠心病等缺血性心脑血管疾病的重要病理基础。中医学“痰瘀互结”是AS主要病理机制，其始动因素表现为脂质沉积。胆固醇逆向转运(RCT)的能力决定AS的进程，而细胞自噬是RCT调节的重要方式。近年报道具有良好成药性的中药单体雷公藤红素（CeT）通过促进RCT抗AS，但机制未明。前期发现，CeT可诱导荷脂细胞发生自噬，故提出“CeT介导的细胞自噬促进RCT”的思路：即CeT靶向PPARγ，转录调控Wnt5a，经PKC/mTOR/TFEB信号通路，诱导细胞自噬，促进RCT。本研究拟建立荷脂细胞及apoE敲除小鼠AS模型，分别在细胞和整体水平观察CeT对自噬和RCT的影响；采用分子对接分析和pull-down技术，鉴定其作用靶点；采用病毒载体构建稳定细胞株，予以小分子干扰或激动剂阐明分子机制。该研究将揭示CeT作为传统中药单体抗AS的潜在靶点和新机制，为中药研发提供新的靶点和思路

Atherosclerosis (AS) is the main pathological basis of ischemic cerebrovascular diseases such as coronary heart diseases. In the concept of traditional Chinese medicine, syndrome of intermingled phlegm and blood stasis is the main pathological mechanism to AS with lipid deposition as the initiative factor. While the imbalance of intracellular cholesterol transport is an essential factor in the development of AS and the capacity of reverse cholesterol transport plays a key role in determining the process and outcome of AS, autophagy is a primary way to regulate intracellular RCT. Recent reports have revealed that celastrol as traditional Chinese medicine monomer exhibits good druggability and it can play the anti-atherosclerosis role by promoting RCT. However, so far the underlying mechanism remains unclear. Our previous study has confirmed that celastrol can enhance autophagy in lipid-loaded cells. Hence our present proposal is based on the hypothesis that celastrol can promote RCT to exert the role of anti-atherosclerosis by activating autophagy. Specifically speaking, we believe that celastrol can target PPARγ/Wnt5a to induce autophagy via PKC/mTOR/TFEB signaling pathway and as a result accelerate RCT. This project will establish lipid-loaded cell and AS mice models, and then focus on observing the effect of celastrol on autophagy and RCT at the cell and the whole level. Thereafter, it will endeavor to identify the underlying targets of celastrol by using molecular docking analysis and pull-down assays. Furthermore, it will clarify the potential mechanism by small molecule interference or agonist in established stable cell lines using retroviral vector infected systems. By accomplishing this proposal, we expect to identify potential targets and new mechanisms for celastrol as a traditional Chinese medicine, to advance the anti-atherosclerosis drug development, and to provide novel approaches in developing traditional Chinese medicines.