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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。
Objective To investigate the dynamic changes of hepatic tissue proteome and the intervention of Fuzhenghuayu Decoction in the formation and depletion of hepatic fibrosis in carbon tetrachloride (CCl4) rats. Methods CCl4-induced hepatic fibrosis model was established by subcutaneous injection of 40% CCl4-olive oil solution. The treatment group was given Fuzheng Huayufang intragastrically. The rats were killed in batches on the 4th, 8th, 12th and 16th weekends. The histopathology and hydroxyproline (Hyp) content in liver tissue of rats were determined. Hepatic tissue proteins were extracted and analyzed by two-dimensional electrophoresis. Protein profiling was analyzed by PDQUEST 2-DE software and differentially expressed by MALDI-TOF-MS. protein. Results (1) Results of two-dimensional electrophoresis: Proteins with different expression levels and protein levels were observed in the model rats from the 4th week and peaked in the 12th week; (2) most of the proteins identified by the mass spectrometry appeared in the 4th week; There were 10 cases in each segment, 8 of which were not found in the normal group; (3) The regulation of protein expression in the Fuzheng Huayu Decoction on the expression differences was as follows: 4th and 8th weeks had significant effect on protein up-regulation. The up-regulation of protein was significantly increased at 8, 12 and 16 weeks. Proteins specific to model rats, after intervention of Fuzheng Huayu Decoction, tend to be normally expressed through degradation, down-regulation of expression and other forms. Conclusion (1) The change of protein expression is the material basis of the overall lesion of liver fibrosis. Abnormal expression of proteins involved in the body’s metabolism, neuroendocrine, cell proliferation and apoptosis systems, (2) Fuzheng Huayufang has comprehensive advantages in anti-cell proliferation, promote normal protein synthesis and other aspects, through multiple proteins, multiple pathways The comprehensive regulation of anti-hepatic fibrosis results.