我国的肝纤维化群体十分庞大，探索防治肝纤维化的有效方法是当前非常必要和迫切的工作。本课题组在上一个国家自然科学基金的研究中，筛选和鉴定到EGCG、Taurine和Genistein三者联合抗肝纤维化的5个候选靶蛋白。本项目将对这5个候选靶蛋白进行功能验证和质谱确认：首先以Taurine、EGCG和Genistein联合作用肝星状细胞和肝纤维化大鼠，采用QPCR和Western-blot技术对候选靶蛋白的mRNA和蛋白质在细胞、血清和组织中的表达进行验证；然后在肝星状细胞中，通过沉默或过表达候选靶蛋白基因进行功能验证，以验证候选靶蛋白与肝纤维发生发展的关系；而后建立以MRM为核心的质谱MIDAS技术流程方案，对候选靶蛋白在大鼠血清及肝组织中的表达进行定性确认；最后采用多通道微流控固相多肽合成芯片技术合成目标多肽，实现MRM技术对靶蛋白在大鼠血清和肝组织中的表达进行最终绝对定量确认。

The hepatic fibrosis people is very large in china. The prevention and treatment of liver fibrosis is currently very necessary and urgent work . The previous project, based on the National Natural Science Foundation (NSFC) , screened and identified five candidate target proteins by iTRAQ labeling and LC-MALDI-TOF/TOF for EGCG, Taurine and Genistein three drug combination against liver fibrosis. This project will confirm target protein candidate by functional verification and mass spectrometry validation. After Taurine, EGCG and Genistein combinate therapy of hepatic stellate cells and in rats, using QPCR and Western-blot analyses mRNA and protein expression of candidate target proteins in cells, serum , and organization. Through gene silencing and overexpression in liver cells, The candidate target proteins are verified the relationship between target protein and liver fibrogenesis. Establish MRM technology as the core of MIDAS process solutions The absolute quantification of the candidate target protein will validated MRM technology as the core of MIDAS process solutions.