抗肝纤冲剂对肝纤维化小鼠Ⅰ型胶原蛋白表达的影响

Influence of Kangganxian Chongji on expression of collagen Ⅰ in hepatic fibrosis tissue of mice

目的:探讨抗肝纤冲剂对猪血清所致的免疫损伤肝纤维化小鼠肝组织Ⅰ型胶原(ColⅠ)蛋白表达的影响,阐明抗肝纤冲剂对肝纤维化小鼠的治疗作用。方法:将75只雌性Balb/c小鼠分为5组,每组15只:正常对照组,肝纤维化模型组,秋水仙碱组,抗肝纤冲剂低剂量组,抗肝纤冲剂高剂量组。除正常对照组外,其余各组均采用腹腔注射猪血清的方法制备肝纤维化小鼠模型。造模的同时分别用药物对小鼠进行干预治疗7周。7周末处死小鼠,取肝组织,经固定、切片、HE染色,光镜下观察。其他肝组织用于实时定量PCR和Western blotting检测。结果:肝组织病理学结果显示,抗肝冲剂治疗组与模型组比较,肝组织的炎症减轻;抗肝纤冲剂低剂量组与肝纤维化模型组比较,肝组织中的ColⅠa2在mRNA表达水平显著降低(P<0.01);Western blotting检测结果显示,抗肝纤冲剂高剂量组的肝组织中ColⅠa1的表达也显著减少。结论:抗肝纤冲剂可以显著减少免疫损伤肝纤维化小鼠肝组织ColⅠ的表达,提示抗肝纤冲剂对肝纤维化小鼠的具有较明显的治疗作用。

Objective To explore the influence of Kangganxian Chongji on the expression of the hepatic collagenⅠ（Col Ⅰ）in the mice with hepatic fibrosis and clarify its theraputical function.Methods 75 female Balb/c mice were randomly divided into 5 groups（n=15）：normal control group,hepatic fibrosis model group,Colchicine group,low dosage therapeutic group and high dosage therapeutic group of Kangganxian Chongji.The hepatic fibrosis model mice were induced by intraperitoneal injection of pig serum respectively（0.5 mL/once,once a week,total 7 weeks）except for normal control group.At the same time the mice in therapeutic group were treated with Colchicine or Kangganxian Chongji once a day,lasted for 7 weeks.The mice were killed and the livers were took from the mice at the end of the 7 th week.The liver tissue slices were made and dyed with HE,and then the pathological changes of the mouse liver were observed under microscope.The mRNA expression of Col Ⅰa2 and protein expression of Col Ⅰa1 in the liver tissues were detected with real time PCR and Western blotting respectively.Results The pathological result indicated that the inflammation of liver tissue in Kangganxian Chongji group was alleviated compared with model group.In low dosage Kangganxian Chongji group,the expression of Col Ⅰα2 mRNA in liver tissue was markedly reduced compared with model group（P〈0.01）;at the same time,the expression of ColⅠα1 protein in liver tissue was markedly reduced according to Western blotting analysis compared with model group.Conclusion Kangganxian Chongji can obviously decrease the expression of ColⅠ protein of the liver tissue in the mice with hepatic fibrosis,and the results suggest that Kangganxian Chongji has obvious therapeutic function on hepatic fibrosis.