Influence Of Cyclic RGD Peptide On Hepatic Stellate Cells Biological Behavior

Liver fibrosis is a liver tissue healing reaction after chronic liver injury which is caused by different reasons.Its pathological features are regeneration accompanied significant fibrogenesis and angiogenesis.Hepatic stellate cells play a central role in fibrogenesis in chronic liver injury.Their activation is the central link of hepatic fibrosis.Activated hepatic stellate cells increase proliferation,migration and mobility,synthesize a large number of extracellular matrix,secrete more pro-angiogenic factors,and induce the expression of integrin αvβ3,which is related to angiogenesis.These changes in biological behavior promote the reconstruction of interstitial.Integrin is a surface transmembrane receptor composed of alpha and beta subunits.It links cells,the extracellular matrix and cell adhesion.Sametimes,it mediates and integrates information transfer of internal and external.Each subtype of the integrin family exhibiting a certain rule of time and the distribution of specific space when expression.Subtype of integrin αvβ3 is the most widely extracellular matrix receptors,which play an important role in the adhesion and information transfer process of activated hepatic stellate cells in contact with the extracellular matrix such as laminin,fibronectin,collagen type Ⅰ,vWF.Moreover,it also has a signal synergistic effect with cell growth factor,but there is no expressing in normal liver tissue and quiescent hepatic stellate cells.RGD cyclic peptide with Specific conformation by artificial synthesis can be recognized by integrin αVβ3 receptor specifically.In order to provide experimental basis for the effect of integrin αVβ3 receptor intervention in anti liver fibrosis treatment,we make the experiment about RGD cyclic peptide intervene on activated hepatic stellate cells to observe its effects on some biological behaviors,such as cell proliferation,migration,mobile and contraction,and αVβ3 expression.Objective:To explore the effects of RGDyK cyclic peptide on integrin αvβ3 expression and cell migration in activated hepatic stellate cells.Methods:Isolation and culture of primary hepatic stellate cell from Sprague-Dawley rat and establish of activated hepatic stellate cells model in vitro,with RGDyK cyclic peptide treatment of activated hepatic stellate cells,by MTT to calculate the growth inhibition rate of cells,by immunochemistry staining and immunofluorescence staining,RT-PCR and Western Blot analysis of integrin αvβ3 and α-SMA expression in activated hepatic stellate cells compared with controls group,observation the effects of cell migration by wound scratche assay.Results:RGDyK cyclic peptide inhibited activate hepatic stellate cells proliferation compared with control group and showed concentration-response relationship,RGDyK cyclic peptide inhibited the expression of integrin αvβ3 and a-SMA both of mRNA and protein in activated hepatic stellate cells respectively(P<0.05),and weaken the mobility of activated hepatic stellate cells(P<0.01).Conclusion:RGDyK cyclic peptide inhibited activated hepatic stellate cell proliferation and migration by down-regulated expression of integrin αvβ3.