扶正化瘀方对肝纤维化大鼠不同病理阶段肝组织蛋白质组变化的影响

目的探讨四氯化碳（CCl4）大鼠肝纤维化形成及消减不同时期肝组织蛋白质组的动态变化及扶正化瘀方的干预作用。方法40％CCl4-橄榄油溶液皮下注射法制备CCl4大鼠肝纤维化模型，治疗组给予扶正化瘀方灌胃，在第4、8、12和第16周末分批处死大鼠，观察各组大鼠肝组织病理学、羟脯氨酸（Hyp）含量变化，并提取肝组织蛋白质进行二维电泳，运用PDQUEST 2-DE软件对蛋白质图谱进行分析，以MALDI—TOF—MS鉴定差异表达的蛋白质。结果（1）二维电泳结果：模型大鼠特有蛋白质和表达量差异的蛋白质从第4周始出现，第12周达顶峰，（2）质谱鉴定的蛋白质多数在第4周出现，四个时间段均出现的有10个，其中有8个是正常组所没有的，（3）扶正化瘀方对表达差异的蛋白质调节表现在：第4、8周对上调的蛋白质降低作用显著，在第8、12、16周对下调的蛋白质的升高作用较显著，模型大鼠特有的蛋白质经扶正化瘀方干预后，通过降解、下调表达量等形式，趋向正常表达。结论（1）蛋白质表达改变是肝纤维化整体病变的物质基础。表达异常的蛋白质涉及机体的物质代谢、神经内分泌、细胞增殖凋亡等系统，（2）扶正化瘀方在抗细胞增生、促进正常蛋白质合成等方面有综合优势，通过对多个蛋白质、多途径的综合调节达到抗肝纤维化的效果。

The effect of Fuzheng Huayu Decoction on the hepatic proteome during the formation and resolution of rat liver fibrosis

Objective To investigate the dynamic changes of the hepatic proteome during the formation and resolution of rat liver fibrosis induced by carbon tetrachloride and the effect of Fuzheng Huayu Decoction on it. Methods Liver fibrosis in Wistar rats was induced by subcutaneous injection of 40% CCl4 in olive oil while the rats in the treatment group received Fuzheng Huayu Decoction. Pathological examination, hydroxyproline content determination and protein extraction of the rats livers were performed at four time points, i.e. at the end of 4, 8, 12 and 16 weeks. After 2-dimensional electrophore-sis (2-DE), the gels were analyzed using the PDQUEST 2-DE image analysis software and differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Results (1) The 2-DE examination showed that the model group-specific proteins and differentially expressed proteins presented started at 4 weeks and reached their peaks at 12 weeks, and this was conformed by MALD-TOF-MS identification. (2) In addition, 18 proteins were present at all time points and another 19 proteins were absent in the model group among those identified by MALD-TOF-MS. (3) Fuzheng Huayu Decoction down-regulated proteins and their expressions increased at 4 and 8 weeks, whereas it up-regulated those which decreased at 8, 12 and 16 weeks. Conclusions (1) The changes in protein expressions, which effect metabolism, neuroendocrine function and cell proliferation, are the basis of liver fibrosis. (2) The anti-fibrotic effect of Fuzheng Huayu Decoction is holistic, specifically because it can decrease the abnormal cell proliferation and can increase the synthesis of normal proteins.