The Influence Of Gbe On Proliferation,apoptosis And Collagenous Protein Synthesis Of Primary Cultured Hsc Detached From Sd Rat

AimsTo investigate the influence of GBE under different concentration on primary cultured HSC of SD rat in its cell proliferation,apoptosis collagenous protein synthesisin the process of culture in vitro.Methods1.The rat HSCs were detached through an improved collagenase-digesting methods.2.The producing capacity and survival rate of HSC may be gained by 0.4% Trypan Blue staining, cellular morphology were observed through microscope and cells were identified as HSC by immunocytochemical stain.3.Passage cells were used to investigated the effect of GBE(0,1,10,100,500μg/ml) on proliferation of HSC though MTT method.4.The cells were divided into five groups:normal control group,. four different concentration group(0,1,10,100,500μg/ml), influence of GBE on cell proliferation were observed through MTT method.5.The cells were divided into five groups:normal control group, four different concentration group(0,1,10,100,500μg/ml),. HSCs apoptosis were detected through ANNEXIN V FITC/PI double staining.6.The cells were divided into five groups:normal control group, four different concentration group (0,1,10,100,500μg/ml), collagenous protein synthesis were detected through ELISA method.Results1.The yield of HSC was 2-5×107 per rat, purity exceed 95% and survival rate exceed 90%.2.GBE had a dose-dependent increased inhibitory effect on cell prolifer- ation in HSC at the concentration of 0,1,10,100,500μg/ml, respectively.Each group has different ratio of inhibition (7.83%,21.5%,32.4%,44.8%,P<0.05)3.The apoptosis rate was gradually increased accompanying the increase of different GBE concentration by ANNEXIN VFITC/PI double staining(P< 0.01)4.ⅣCollagenous protein synthesis was gradually decreased accompanying the increase of different GBE concentration by ELISA method.HA synthesis was gradually decreased accompanying the increase of different GBE concen-tration by ELISA method(P<0.01)Conclusion1.The rat HSCs were detached through an improved collagenase-digesting method successfully.It is an important model to use primary cultured rat HSC to investigate liver fibrosis in vitro.2.GBE can inhibit HSC proliferation and accelerate its apoptosis obviously and it can decrease the content of HA and IVcollagen.