CCl_4大鼠肝纤维化形成与消减过程中肝组织蛋白质组的动态变化及扶正化瘀方的干预作用

Dynamic Demonstration on Hydrophilicity Protein Molecular of Fibrosing Hepatic Tissue in Rats with Intervention of Fuzheng Huayu Decoction

目的:通过大鼠纤维化肝组织高亲水性蛋白质分子差异展示及扶正化瘀方干预作用,寻找与扶正化瘀方治疗肝纤维化相关的蛋白质分子。方法:通过皮下注射大鼠CCl4,分别持续4、8、12周,对大鼠肝纤维化过程进行动态观察,并在不同时期进行扶正化瘀方干预,治疗从第12周始至16周止。采用蛋白质分步抽提方法,提取肝组织蛋白质,分别进行二维聚丙烯酰胺凝胶电泳(2DE)分离、凝胶银染,显示各组蛋白质分子的差异表达的蛋白质分子。结果:在各组银染凝胶图谱上,可辨识的蛋白质斑点达1000～1400个左右,蛋白质表达中除有肝组织看家蛋白质,模型组也有特异蛋白质表达,且在不同时期蛋白质分子表达也有差异,特异表达蛋白质分子为100至300多个不等,在不同阶段表达是不相同的,经质谱鉴定确定了有几类功能不同的蛋白质分子。结论:扶正化瘀方抗肝纤维化作用机制是多方面的,其干预作用与多种蛋白质分子相关。

Objective:By demonstration of the differences in hydrophile protein molecular of hepatic tissue in rats, correlative protein molecular to intervention in hepatic fibrosis with Fuzheng Huayu decoction has been observed to seek. Methods: Injection of CCl 4 hypodermicly last for 4、8、12 weeks separately to make a dynamic animal model of hepatic fibrosis in rats and therapeutic intervention to them from the week 12th to 16th. Proteins in hepatic tissue were extracted by methods of protein extraction step by step and separated by 2-DE(2 dimensional electrophoresis) to detect the differential protein molecular among varied groups by silver stain. Results: According to gel maps among varied groups, approximately 1 000 to 1 400 protein spots was visible. Family proteins were observed in hepatic tissue and special proteins were observed in model group, in addition, special proteins varied from time to time. The differential proteins were about several deca to one hundred. Molecular weight and equipotentiality have been confirmed primarily in the course of the study. Conclusion: There are multi-aspects to the mechanism of anti-hepatic fibrosis with Fuzheng Huayu decoction and its intervention effects could be related to varied proteins.