芪蚣抗纤方拆方对肝纤维化大鼠Smad2/3及Smad7蛋白的影响

Impact of Qigong Kangxian Prescription on Smad2/3 and Smad7 in Hepatic Fibrosis Rat

目的:观察芪蚣抗纤方(Qigong Kangxian Prescription,QF)活血组和软坚组对肝纤维化大鼠细胞内信号转导分子2,3,7(Smad 2/3及Smad7)的影响。方法:成年健康雄性SD大鼠60只,随机分为6组,除正常对照组10只外,其余50只均用猪血清腹腔注射法复制免疫损伤性肝纤维化大鼠模型,0.5 mL/只,每周2次,连续8周。活血组和软坚组分别给于活血方和软坚方。低、高剂量组按生药量以5,10 g.kg-1ig给药。实验结束处死大鼠,断头取血,分离血清,用ELISA法测定转化生长因子(transforming growth factor,TGF)β1的含量。部分肝组织固定于10%甲醛液,石蜡包埋,制光镜切片,免疫组织化学法检测肝组织TGF-β1、Smad2,Smad3及Smad7蛋白的表达。结果:①血清及肝组织TGF-β1含量及表达活血高剂量组分别为(0.65±0.09)μg.L-1,4.33±2.06,软坚低剂量组分别为(0.62±0.08)μg.L-1,4.11±1.14,与模型组(1.22±0.16)μg.L-1,10.33±3.34相比有显著差异(P<0.01);②Smad2和Smad3蛋白表达显色指数活血高剂量组分别为(3.88±2.00),(3.22±1.89)分,软坚低剂量组分别为(4.22±1.49),(4.77±1.75)分,与模型组(10.20±3.06),(10.22±3.06)分相比有显著差异(P<0.01);③Smad7蛋白表达显色指数活血高剂量组为(6.89±1.50)分,软坚低剂量组为(4.33±1.78)分,与模型组(1.56±0.73)分相比有显著差异(P<0.01)。结论:芪蚣抗纤方活血组及软坚组可通过减少Smad 2/3的激活,增加Smad7的表达,抑制肝纤维结缔组织增生,减轻肝损伤及肝纤维化程度。具有抑制肝纤维化作用。

Objective： To explore the impact of Qigong Kangxian prescription（QF） on Smad 2/3,Smad7 in hepatic fibrosis rats.Method： Sixty SD rats randomly divided into 6 groups,in addition to the normal control group,the other groups were intraperitoneally injected with 0.5 mL pig＇s serum twice per week for 8 weeks,the rat hepatic fibrosis model was induced.Rats were given corresponding herbal medicines.After the end of experiment,serum samples were collected.Determination of the content of transforming growth factor was carried out by using ELISA method.Sections of liver tissue was fixed in 10% formaldehyde solution,and then embedded in paraffin,Under the light microscope.Expression of liver tissue TGF-β1,Smad2,Smad3,Smad7 was detected by immunohistochemical method.Result： In the Content of TGF--β1 and expression of liver tissue TGF-β1,Smad2,Smad3 and Smad7 the significant differences were found in administration groups compared with the model group（P 0.01）.Conclusion： QF can suppress hyperplasy of hepatic fibrous connective tissue,lessen hepatic injury through decreasing Smad 2 /3 expression and increase Smad7 expression.