Experiment Study Of5-FU On Role Of Human Bile Duct Scar Fibroblasts In Vitro

Objective:To explore the role of5-FU of Human benign bile duct scar fibroblasts in vitro.Methods:1.The resuscitation and serial subcultivation of human benign bile duct scar fibroblast.2.Different concentration5-FU treatment of human benign bile duct scar fibroblast; human benign bile dust scar fibroblast; human benign bile dust scar fibroblast were stimulated with5-FU(0.78125g/l,1.5625g/l,3.125g/l,6.25g/l,12.5g/l);Different concentration5-FU treatment of human benign bile duct scar fibroblast for24,48,72Hours, the human benign bile duct scar fibroblast is observed under the100times Optical microscope and the inhibition ratio and50%Inhibitiory concentration(IC50) were calculated by MTT.In addition, the most approach IC505-FU concentration were prepared for the following experiment.3. The cell apoptosis were measured by flow cytometry.4.The influence of5-FU on TGF-β1,a-SMA,collagen-Ⅰ and collagen-Ⅲ expressions were detected by Immune fluorescent chemical.5.The influence of5-FU on TGF-β1,a-SMA,collagen-Ⅰ and collagen-Ⅲ expressions were detected by Western Blot.Results:1.5-FU treatment of human benign bile duct scar fibroblast for72hour, IC50is6.25g/L;2.After intervention of Different concentrations of5-FU in the bile duct scar fibroblasts, the growth curves show that cells are inhibited more with time and the concentration of drug increasing,72hours suppression effect is the most obvious(see Figure10);3.5-FU treatment of human benign bile duct scar fibroblast, its apoptosis obvious greater than blank group;4.5-FU treatment of human benign bile duct scar fibroblast in vitro, TGF-β1, a-SMA,collagen-Ⅰ and collagen-Ⅲ expressions were detected downgrade by Immune fluorescent chemical and Western Blot.Conclusions:1.5-FU ihibits the growth of the human bile duct scar fibroblasts2.5-FU inhibits the growth of the human bile duct scar fibroblasts by the promotion of apoptosis.3.After the intervention of5-FU in human bile duct scar fibroblasts, the TGF-β1, a-SMA, type Ⅰ collagen, type Ⅲ collagen expression is significantly reduced.4.The research revealed the mechanism of action on5-FU treated human benign biliary scars stricture.5-FU could induce apoptosis on human benign bile duct scar fibroblast in vitro. With5-FU topical administration would be a new consider of human benign biliary scars stricture treatment.