中药肝复康干预肝星状细胞及肝组织的蛋白质组学分析

目的：本实验通过中药对肝星状细胞和肝组织蛋白质组的干预实验，从蛋白质组学角度进一步揭示肝纤维化的发病机制和中药的药理作用，为新药研制提供理论依据。方法：制备正常大鼠肝星状细胞、加入血小板衍生生长因子的肝星状细胞及加入肝复康药物血清和血小板衍生生长因子的肝星状细胞，分别设定为正常对照组、模型组及治疗组；同时分别将注射二甲基亚硝胺、肝复康的大鼠设定为模型组及治疗组，并将各组大鼠肝脏取出。分别提取以上各组的总蛋白质并进行等电聚焦电泳，随后进行SDS-PAGE电泳。凝胶染色后比对蛋白质斑点，找出上调和下调的蛋白质。结果：随着治疗时间的不断延长，HSC的蛋白质表达在不同时间有了不同变化。大鼠造模4周后，模型组与治疗组肝组织总蛋白质图谱存在着一定差异，部分蛋白质在两组中表现出不同的丰度。结论：肝纤维化的发生与多种蛋白质的作用有关。中药肝复康治疗肝纤维化是通过调节多个蛋白质表达产生的。

Proteomics Analysis of Effect of Chinese Traditional Medicine Ganfukang on Liver Stellate Cells and Tissue

Objective： To investigate the influence of Chinese medicine on liver tissues and HSC by 2-DE and to supply the theory basement for drug development. Methods： HSCs were assorted for three groups： control group, model group, Chinese herb-treated group; Live tissues were divided into model group and Chinese herb-treated group. All protein from those groups was extracted. The method of O＇Farrell, with modification, was followed. Isoelectric focusing （IEF） separation was carried out with a vertical mini-IEF system （Jim-X Company） and the SDS-PAGE was carried out to separate proteins in the second dimension. Results： There are different protein spots in different phase comparing Gel scan among three model groups both in HSCs and liver tissues. The number of protein is changed as the time prolonged.After building rats model 4 weeks, there existed some differences between model group and Chinese herb-treated group in total protein atlas. Some proteins shows different abundance in different groups. Conclusions： There is significant difference at protein level among different groups. This shows that the pathogenesis of liver fibrosis involve in many kinds of protein. It suggests that the differential expression of proteomes were useful to further study on mechanism of Ganfukang, and search for new targets of anti-liver fibrosis medicine.