补肾活血化浊方对Wilson病TX小鼠肝纤维化肝再生作用及机制研究

Liver regeneration effect and mechanism of Bushen Huoxue Huazhuo Recipe on Wilson disease liver fibrosis of TX mice

目的:观察补肾活血化浊方对Wilson病TX小鼠肝组织转化生长因子-β1(TGF-β1)、骨形态蛋白-7(BMP-7)的影响,探讨其对Wilson病TX小鼠肝纤维化肝再生的作用机制。方法:将40只TX小鼠随机分为模型对照(MG)组、中药补肾活血化浊方(TCM)组、青霉胺(PCA)组和谷胱甘肽(GSH)组,每组10只,另设DL小鼠10只为空白对照(NG)组,采用羟脯氨酸(Hyp)试剂盒检测小鼠肝组织Hyp含量,采用RT-PCR法和Western Blot法分别检测肝组织中TGF-β1、BMP-7 mRNA和蛋白的表达。结果:与NG组比较,MG组TX小鼠肝组织Hyp含量显著升高(P<0.01);与MG组比较,各给药组TX小鼠肝组织Hyp含量显著降低(P<0.01)。与NG组比较,MG组TX小鼠TGF-β1 mRNA和蛋白表达显著升高(P<0.01),BMP-7 mRNA和蛋白表达降低(P<0.05,P<0.01);与MG组比较,各给药组TX小鼠肝组织中TGF-β1 mRNA和蛋白表达均显著降低(P<0.05,P<0.01),BMP-7 mRNA和蛋白表达均升高(P<0.01)。结论:补肾活血化浊方对Wilson病TX小鼠肝纤维化具有抑制作用;其机制可能是通过促进Wilson病TX小鼠肝组织中BMP-7表达,抑制TGF-β1表达,减少胶原纤维生成,以此调节肝再生过程发挥抗肝纤维化作用。

Objective:By observing the effect of Bushen Huoxue Huazhuo Recipe on TGF-β1,BMP-7 of liver issue of TX mice,to investigate the mechanism of liver regeneration in Wilson disease TX mice with liver fibrosis.Methods:A total of 40 TX mice were randomly divided into model control(MG)group,traditional Chinese medicine Bushen Huoxue Huazhuo Recipe(TCM)group,Penicillium(PCA)group and glutathione(GSH)group,10 DL mice were as the blank control(NG)group.Finally,the content of hydroxyproline(Hyp)in the liver tissue of mice was detected by hydroxyproline kit,the mRNA expression of TGF-β1 and BMP-7 in the liver tissue was detected by RT-PCR,and the protein expression of TGF-β1 and BMP-7 in the liver tissue was detected by Western Blot.Results:The Hyp content in the MG group was significantly higher than the NG group(P<0.01);Compared with model control group,the content of Hyp in liver tissue of TX mice in three giving drug groups decreased significantly(P<0.01).Compared with NG group,the expression of TGF-β1 mRNA and protein in the liver tissue of TX mice of the MG group was significantly higher(P<0.01),the expression of BMP-7 mRNA and protein in the MG group was significantly lower(P<0.05,P<0.01).Compared with the MG group,the expression of TGF-β1 mRNA and protein in the liver tissue of TX mice in three giving drug groups were significantly lower(P<0.05,P<0.01),while their BMP-7 mRNA and protein were significantly higher(P<0.01).Conclusion:Bushen Huoxue Huazhuo Recipe has inhibitory effect on liver fibrosis of TX mice with Wilson’s disease,which may be to promote the expression of BMP-7,inhibit the expression of TGF-β1,and reduce the generation of collagen fibers,so as to regulate the process of liver regeneration and play an anti fibrosis role.