纤维化大鼠肝组织与物质代谢相关蛋白质动态变化及扶正化瘀方的调节作用

Dynamic Change of Metabolism Related Protein in Liver Tissue of Rats' M odel of Hepatic Fibrosis and Regulatory Effect of Fuzheng Huayu Decoction on It

目的探讨扶正化瘀方干预治疗肝纤维化的作用。方法将Wistar大鼠随机分正常组、模型组、扶正化瘀方(简称中药)组。大鼠皮下注射40%CCl4-橄榄油溶液造模,正常组仅用生理盐水皮下注射,中药组用中药灌胃,取肝组织做病理、羟脯氨酸、蛋白质的分步提取。蛋白质定量后进行二维电泳,凝胶银染,运用PDQUEST2-DE图像分析软件对获得的蛋白质图谱进行分析,运用MALDI-TOF-MS鉴定了30多个差异表达的蛋白质。结果(1)模型组大鼠出现生命活动整体反应(如体重、活动度等)下降;(2)肝纤维化过程中存在物质代谢障碍;(3)蛋白质组改变质谱结果显示模型组和正常大鼠肝组织差异表达的蛋白质与物质代谢相关的蛋白质有:过氯酸可溶性蛋白质、磷脂酰肌醇转移蛋白酶、磷酸甘油酸激酶、内质网-60蛋白酶;(4)中药组大鼠肝组织中过氯酸可溶性蛋白质、磷脂酰肌醇转移蛋白酶、磷酸甘油酸激酶、内质网-60蛋白酶的表达趋近正常。结论在肝纤维化过程中伴有蛋白质合成与分解异常和糖酵解活动增强;中药调节物质代谢相关蛋白质的表达是抗肝纤维化的途径之一。

Objective To investigate the effect of Fuzheng Huayu decoction （FHD） intervention on hepatic fibrosis. Methods Wistar rats were randomly divided into 3 groups： rats in the normal group only treated with subcutaneous injection of saline, rats in the model group and the FHD group were made into hepatic fibrosis by subcutaneous injection of 40 % carbon tetrachloridc （CC14）-olive solution and then those in the FHD group were treated with FHD by gastric perfusion after modeling. Liver samples of the rats were obtained for routine pathological observation, hydroxyproline determination and proteomc quantitative determination. After then, the proteome profile was obtained through 2-dimensional electrophoresis and silver staining, and analyzed, More than 30 proteins with different expression were identified by MALDI-TOF-MS. Results （1） The integral response of vital movement such as body weight and activity of hepatic fibrosis declined in the CCl4 induced liver fibrosis rats; （2） Liver fibrosis were associated with abnormal metabolism; （3） There were four material metabolism-related protein showed by hepatic proteome mass spectrography, which expressed different between the normal and the fibrotic rats, i. e, the perchloric acid soluble protein, the phosphatidylinositol transferase, the phosphoglycerate kinase and the endoplasmic reticulum-60 protease; （4） The expressions of the above-mentioned four proteins in the FHD group were nearly the same as those of normal level. Conclusion （1） Liver fibrosis is accompanied with abnormal protein synthesis and decomposition, as well as the enhanced activity of glycolysis; （2） The existence of metabolism-related proteins is one of the elements for the liver in regulating metabolism; （3） The regulation on the expressions of metablism-related proteins is one of the pathways for FHD to exert its anti-hepatic fibrosis effect.