复方鳖甲软肝片治疗免疫性肝纤维化大鼠肝细胞质膜蛋白质组分的变化

目的研究复方鳖甲软肝片治疗肝纤维化大鼠肝细胞质膜蛋白质组分的变化，以寻找与肝纤维化逆转相关的质膜蛋白质。方法以猪血清腹腔注射大鼠8周制备肝纤维化模型，再以复方鳖甲软肝片灌胃治疗8周。采用蔗糖密度梯度离心获得肝组织匀浆和质膜组分，采用免疫印迹法检测质膜的纯度，分析差异蛋白质组学变化。结果病理学检查发现实验8周后，大鼠肝脏发生中度肝纤维化；经鳖甲软肝片治疗8周后肝纤维化程度减轻，基本恢复到正常水平；免疫印迹检测显示质膜组分得到了较好的富集，其中线粒体等其他细胞器蛋白减少；通过差异蛋白质组学分析发现，治疗组中有22个3倍以上差异蛋白质点，准确鉴定了9个蛋白质，通过免疫印迹验证发现了Ⅱ型细胞支架蛋白8和膜联蛋白A2的差异表达。结论鳖甲软肝片治疗后，大鼠肝纤维化发生了逆转，并伴随一些质膜蛋白质的表达变化。

Differential proteomic analysis of liver plasma membrane in hepatic fibrosis rata treated with Fufang Biejia Ruangan Pian

Objective To probe the membrane protein changes inrats with hepatic fibrosis after the treatment of Fufang Biejia Ruangan Plan. Method This research set up an rat model with porcine sera-induced hepatic fibrosis and the model rats were treated by intragastric infusion of Fufang Biejia Ruangan Plan solution for 8 weeks. Plasma membrane was purified by density gradient centrifugation,the purity of them was verified by Western blot and differential proteomic analysis of liver plasma membrane was done. Result An animal model of hepatic fibrosis was set up successfully at week 8. After treatment with Fufang Biejia Ruangan Pian for eight weeks,the livers returned to nornlal levels;Western blot analysis showed that the plasma membrane was enriched while the contamination of other subcellular fractions such as mitochondrion reduced. 22 proteins with more than three-fold differences were found and 9 proteins were successfully identified. Western blotting verified the different expression of keratin 8 and annexin A2. Conclusion Fufang Biejia Ruangan Plan has anti-fibrotic effects by regulating keratin 8 and annexin A2 expression in the plasma membrane during hepatic fibrosis process.