| 【Objective】To improve the theory of TCM about collateral disease,and summarized the law of hepatic collateral disease treatment,to explore the traditional Chinese medicine compound Dahuangzhechong-pill combined with Astragalus and Fructus Polygoni orientalis regulating liver meridian function,and to explore the anti fibrosis effect of Dahuangzhechong-pill after and the regulation mechanism of MMP-1/TIMP-1 in the liver from modern medicine pathology perspective study on whether it can enhance the efficacy of anti hepatic fibrosis.To provide theoretical basis and experimental basis for the diagnosis and treatment of hepatic fibrosis.【Methods】 Review of the medical classics of Troditional Chinese Medicine theory in-depth,classic literature research,and summarizes the back liver meridian theory origin,to provide a theoretical basis for experimental research.An experimental study on porcine serum model,immune hepatic fibrosis model in SD rats induced by porcine serum,respectively set up Normal group,Model group,Dahuang Zhechong-Pill group,Jiawei Dahuangzhechong-pill group,Colchicine group,through 12 weeks of feeding and injection molding,6 weeks of intragastric administration,then collected serum samples and tissue samples,the liver and spleen index,serum liver function and liver fibrosis markers in liver tissue were observed by HE staining(hematoxylin-eosin staining)and MASSON staining pathology,expression of messenger RNA of MMP-1(metalloproteinase-1)and TIMP-1(tissue inhibitor of metalloproteinase-1)in liver tissue were detected by RT-PCR(real-time fluorescence quantitative PCR polymerase chain reaction).Elisa(enzyme immunosorbent assay,enzyme-linked immunosorbent assay)method was used to detect the changes of MMP-1 and TIMP-1 protein in liver tissue of rats.【Results】 1.The general situation: Compared with the normal group,the rats in the model group,the general situation of the spirit,diet,hair color and so on are poor,weight gain is slow.Compared with the model group,the symptoms of the rats in each treatment group were alleviated.2.Rat liver and spleen index: Compared with the normal group,the liver weight of the model group increased,the difference was not significant;spleen weight,liver index and spleen index increased significantly,the difference was significant(P<0.01).Compared with the model group,the liver weight and liver index of the rats in each treatment group were decreased,the difference was not significant,the spleen weight and spleen index were decreased,and the difference was significant(P<0.05).3.Liver pathology ofrats: Observation of liver pathology under light microscope,HE staining showed that the structural damage of liver lobule rats in the model group,the liver cells arranged in disorder,visible ballooning degeneration,periportal enlargement accompanied by inflammatory cell infiltration and collagen formation of fibrous septa,the size range of pseudolobuli;MASSON trichrome staining showed that collagen of the rats in the model group liver fiber thickening,formation of fibrous septa have obvious formation of pseudolobuli.Colchicine group,Dahuangzhechong-pill group and Jiawei Dahuangzhechong-pill group,the liver cells arranged more orderly,abnormal structure of liver cell is rare,a small amount of collagen fibers deposition,inflammation and fibrosis of liver was slightly reduced compared with model group(P<0.05).4.Liver function : Compared with normal group,model group,serum Ast(aspartate aminotransferase,aspartate aminotransferase)were significantly increased,significant difference(P<0.05);Alt(alanine,aminotransferase,serum alanine aminotransferase(gamma-glutamyl),Ggt transpeptidase,r-glutamyltranspeptidase(total Tbil),bilirubin,total bilirubin elevated serum Tp(total),protein,total protein(albumin),Alb,albumin,Glb(globulin),immunoglobulin(albumin/),A/G globulin,white ball ratio)decreased,the difference was not significant.Compared with the model group,the serum Alt,Ast,Ggt decreased,Tbil increased,serum Tp,A/G decreased,serum Glb and Alb increased,the difference was not significant.Compared with the model group,Dahuang Zhechong Pill on serum Alt,Ast and Tbil decreased,there were significant differences(P<0.05);the serum Ggt decreased,serum Tp,Glb,Alb and A/G increased,the difference was not significant.5.Serum markers of liver fibrosis: Compared with the rats in the normal control group,model control group rats serum HA(hyaluronic acid),LN(laminin),and PC III(procollagen type III),IV-C(type IV-collagen)increased significantly(P<0.05);colchicine group and Dahuangzhechong pill group HA,LN,PC III compared with the model control group was significantly decreased(P<0.05);Jiawei Dahuangzhechong-pill group HA,LN,IV-C,PCIII was significantly lower than the model control group(P<0.05).6.The expression of mRNA of MMP-1/TIMP-1 in liver tissue : Compared with normal group,the liver fibrosis model group increased,MMP-1m RNA decreased and TIMP-1m RNA increased(P<0.05,P<0.01);compared with the model group,Dahuangzhechong pill group,Jiawei Dahuangzhechong pill group MMP-1m RNA had increased trend,TIMP-1m RNA had a decreasing tendency,which the difference of Jiawei Dahuangzhechong pill group significantly(P<0.01).7.The content changes of MMP-1/TIMP-1 protein in liver tissue: Compared with normal group,MMP-1 model group rats decreased,TIMP-1 increased,the difference was statistically significant(P<0.05 or P<0.01);compared with the model group,colchicine group,Chinese medicine group TIMP-1 decreased,the difference has statistical significance(P<0.05).【Conclutions】 From the point of view of traditional Chinese medicine,it can be concluded that "qi stagnation of liver qi" and "blood stasis of liver collateral" are the basic pathogenesis of the formation and development of hepatic fibrosis.The results showed that the pig serum induced liver fibrosis in rats was impaired liver function,fibrous hyperplasia,there were obvious pathological damage of liver tissue,various types of collagen deposition in liver tissue,resulting in MMP-1/TIMP-1 loss of balance.That Jiawei Dahuangzhechong-pill from the modern perspective of Pathology(i.e.Dahuangzhechong-pill combined with Astragalus and Fructus Polygoni orientalis)can regulate MMP-1/TIMP-1 metabolism,inhibit collagen deposition,promote fiber degradation,improve liver function,has a definite therapeutic effect on liver fibrosis,provide some experimental evidences for clinical diagnosis and treatment of liver fibrosis can. |
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