Study On Dihydromyricetin Of Ampelopsis Grossedntata Regulating TGF-β1/Smad And PDGF Signaling Pathway Against Liver Fibrosis

Objective:Based on the signaling pathways of TGF-beta 1/Smad and PDGF,the anti-fibrotic mechanism of Dihydromyricetin（DIH）was studied.Methord:98 KM mice,half male and half female,were randomly divided into six groups:normal control group,model group,colchicine positive control group and DIH high,medium and low dose group.Each groups were induced by subcutaneous injection of 25%CCL₄peanut oil solution in mice once every 3d and for 10 weeks,2 mL/kg,except for the blank control mice.The mice were treated by intragastric administration the corresponding drugs or distilled water once a day and last for 10weeks.At the end of the 10th week,after the last administration of 1 hour,the eyeballs of mice were taken for blood collection,serum was separated and stored in a cryogenic refrigerator for reserve.Liver tissues were harvested and fixed with 4%tissue and cell stationary fluid,and some of them were preserved in cryogenic refrigerator.Respectively test the content of ALT,AST in blood serum and the COL Ⅰ,COL Ⅲ,HA,LN in mice liver.Making pathological section and use HE,Masson staining to see the stage of mice liver fibrosis.The effects of DIH on the expression of major genes and proteins in the TGF-beta 1/Smad and PDGF pathways in liver tissues of mice with hepatic fibrosis were detected by RT-PCR and Immunohistochemistry.Results:Compared with the model group,the high,medium and low dose groups of DIH could reduce the levels of ALT,AST in serum and COL I,COL Ⅲ,HA,LN and Hyp in liver tissue of mice with hepatic fibrosis induced by CCL₄,improve the degree of pathological damage,reduce the infiltration of inflammatory cells in liver tissue,reduce the production of collagen in liver,and reduce the Ishak score.Compared with the model group,The high-dose of DIH can reduce the genetic expression of tgf-β1、smad 2、α-sma The mid-dose of DIH can reduce the genetic expression of smad 2、smad 4、α-sma.The low-dose of DIH can reduce the genetic expression of smad 4、α-sma.The high-,mid-dose of DIH can increase the genetic expression of smad 7.The high-,mid-.low-dose of DIH can reduce the protein expression of TGF-β1、Smad 2、Smad 4.The mid-,low-dose of DIH can reduce the protein expression ofα-SMA.The high-,mid-dose of DIH can increase the protein expression of Smad 7.Compared with the model group,The high-,mid-dose of DIH can reduce the genetic expression of pdgf,pi3k,akt andα-sma.The low-dose of DIH reduce the genetic expression of akt,α-sma.The high-dose of DIH increase the genetic expression of pten.The high-,mid-,low-dose of DIH can reduce the protein expression of PDGF,PI3K,increase the protein expression of PTEN.The mid-,low-dose can reduce the protein expression ofα-SMA.Conclusion:（1）DIH can significantly interfere with liver fibrosis induced by CCL₄.（2）The anti-fibrosis mechanism of DIH may be related to the signaling pathways of TGF-beta 1/Smad and PDGF.