分泌蛋白是药物治疗的有效靶点，是进行疾病诊断、预后和跟踪等生物标志物的丰富资源。分泌蛋白质组的研究有助于找到直接与特定生理或病理状态相关的生物分子。细胞培养稳定同位素标记（SILAC）被认为是目前体外研究分泌蛋白质组最好的方法之一。本研究基于SILAC的分泌蛋白质组学技术，全面阐述中药绞股蓝总苷在治疗肝损伤进程中的作用机理。研究包括（1）建立体外肝损伤及肝细胞凋亡模型，应用SILAC技术进行标记，通过质谱技术、蛋白质数据库查询、生物信息学等方法鉴定差异蛋白表达谱以此建立分泌蛋白组学研究框架；（2）建立体内肝损伤大鼠模型并给予中药绞股蓝总苷治疗，利用免疫印迹、免疫组化等方法对候选关键差异蛋白进行定量研究，以期发现疾病的相关蛋白、标记分子、药物靶标和发病机理。本项目的实施，将有利于分析化学、生物化学、分子生物学等相关学科的衔接与交叉集成，可从微观角度上获取关于细胞代谢、疾病发生等过程整体认识。

Secreted protein is an effective target of drug therapy and a rich resource of biomarkers for disease diagnosis, prognosis and tracking. The study of the secretory proteome helps to identify biomolecules directly associated with specific physiological or pathological states. Stable isotope labeling (SILAC) in cell culture is considered as one of the best methods to study secreted proteome in vitro. Based on the secretory proteomics of SILAC, this study comprehensively elaborated the mechanism of gypenoside in the treatment of liver injury. The research included (1) establishing in vitro models of liver injury and liver cell apoptosis, using SILAC technology for labeling, identifying differential protein expression profiles through mass spectrometry, protein database query, bioinformatics and other methods to establish a framework for secretory proteomics research; (2) a rat model of liver injury in vivo was established and gypenoside was given as treatment. Immunoblotting, immunohistochemistry and other methods were used to conduct quantitative research on candidate key differentially expressed proteins, so as to find the relevant proteins, marker molecules, drug targets and pathogenesis of the disease. Project implementation, will be conducive to analytical chemistry, biochemistry, molecular biology and other related disciplines of cohesion and cross integration, can be obtained from microcosmic point of view about the process of cell metabolism, disease, and so on overall understanding, to reveal the secretory protein in liver injury and liver cell apoptosis process with nature and traditional Chinese medicine (TCM) in the process of gynostemma total glycosides treatment of liver damage potential molecular targets.