Theory Of Gan Luo Bing (Collateral Disease Of Liver) Based On Experimental Research On Effects Of Kangfuxinye On Hepatic Fibrosis Rats

Objectives1.To research theoretical understandings that hepatic fibrosis pertains to Gan Luo Bing?collateral disease of liver?in Traditional Chinese Medicine.2.To research theoretical understandings that Kangfuxinye treats hepatic fibrosis by unblocking the collaterals and vessels.3.By exploring anti-hepatic fibrosis effects and action mechanisms of Kangfuxinye on CCl₄-induced hepatic fibrosis rats,experimental evidences will be provided to strengthen theoretical understandings that hepatic fibrosis pertains to Gan Luo Bing,and to expand applications of Kangfuxinye to treat hepatic fibrosis.Methods1.Theoretical researches:?1?Put forward theoretical understandings that hepatic fibrosis pertains to Gan Luo Bing by literature review on correlations between Gan Luo Bing and hepatic fibrosis.?2?By reviewing researches of Kangfuxinye,theory was proposed that Kangfuxinye unblocks collaterals and vessels to treat hepatic fibrosis due to collaterals of the liver obstructed by blood stasis.2.Experimental researches:?1?The hepatic fibrosis rats were induced by intraperitoneal injection of CCl₄?CCl₄:olive oil=2:3?.?2?Observe the general signs of rats with the naked eye.?3?The pathological changes of liver was observed by HE staining,and the collagen fibers were identified by Masson's trichrome staining.?4?ALT,AST,HA,LN,P?NP,ColIV and Hyp in serum were tested by ELISA.?5??-SMA,Col?and TIMP-1 in the liver were detected by immunohistochemistry.?6?Proteins including TGF-?1,Smad3,?-catenin,Axin and GSK-3?in the liver were detected by Western Blot.Results1.The therapeutic effects of Kangfuxinye:?1?General signs:Rats in control group have shiny hairs,strong physique,vitality and good appetite.While rats in CCl₄treated groups are quite,fatigue with lusterless hair,poor appetite and loose stools.?2?Histology of the liver:Results of HE staining shown that the liver of rats of control group were normal,characterized by clear hepatic lobule,well-arranged hepatic cords,hepatocytes with uniform size and clear nucleus and cytoplasm,instead of degeneration or necrosis,and without hyperplasia of fibrous connective tissue and bile duct.The liver of rats of vehicle treated group manifested as unclear architecture or lesion of liver lobule,disordered hepatocytes with diffuse steatosis,hyperplasia of fibrous connective tissue around portal area.Compared with vehicle treated group,conditions of treatment groups were improved.Results of Masson's trichrome staining shown that the liver of vehicle treated group manifested as massive distribution of collagen fibrils around the lobule,portal area and central vein.Collagen fibrils of the liver of treatment groups were reduced by comparison with vehicle treated group.?3?Results of ELISA tests shown that activity of AST and ALT of treatment groups is reduced compared with vehicle treated group.?4?Results of ELISA tests shown that content of HA,LN,ColIV,P?NP and HyP in serum of treatment groups are lower compared with vehicle treated group.?5?Results of immunohistochemistry shown that?-SMA,Col?and TIMP-1 in the liver of treatment groups are lower compared with vehicle treated group.2.Mechanisms of anti-hepatic fibrosis of Kangfuxinye:Results of Western Blot shown that,compared with vehicle treated group,proteins in the liver including TGF-?1,Smad3 and?-catenin of treatment groups were reduced,while Axin and GSK-3?were increased.Conclusions1.Theoretical researches:TCM holds that invasion of damp-heat and epidemic pathogenic factor causes deficiency of Vital qi failing to promote blood circulation and resulting in blood stasis,then combination of dampness,toxins and blood stasis blocks liver collaterals to cause liver fibrosis,which indicates that liver fibrosis pertains to Gan Luo Bing.2.Experimental researches:?1?Theoretical understandings that hepatic fibrosis pertains to Gan Luo Bing is verified on CCl₄-induced hepatic fibrosis rats by evidences including histology of the liver,fibrotic markers in both serum and the liver,and key proteins of TGF-?1/Smad3 and Wnt/?-catenin signaling pathway.?2?Kangfuxinye could relieve hepatic fibrosis by improving conditions of damaged liver lobule,reducing hyperplasia of fibrous connective tissue,and lowering content of HA,LN,ColIV,P?NP,HyP in serum and?-SMA,Col?,TIMP-1 in the liver.?3?Kangfuxinye could down-regulate TGF-?1,Smad3,?-catenin and up-regulate Axin and GSK-3?to relieve hepatic fibrosis.