Paclitaxel Effect On Fibroblast Of Human Benign Bile Duct Scar In Vitro

[Objective]Explore paclitaxel benign bile duct scar fibroblasts sensitivity study of paclitaxel on human benign biliary hypertrophic scar fibroblasts proliferation,apoptosis and cell cycle.Study of paclitaxel on the expression of typeⅠ、Ⅲcollagen、TGF-β₁andα-SMA find new ways implications for the clinical treatment of bile duct scar.[Methods]The established early recovery and subcultured benign biliary hypertrophic scar fibroblasts,using different concentrations（170mg/L,85mg/L,42.5mg/L,21.25mg/L,10.625mg/L,5.3125mg/L,2.65625mg/L）paclitaxel to intervene at different times（24h,48h,72h）,by morphological observation and MTT assay study of paclitaxel on benign bile duct scar fibroblast proliferation and viability;flow cytometry（TUNEL method）detection paclitaxel 50%inhibitory concentration（IC50）for 48 hours bile duct scar fibroblast apoptosis;immunofluorescence detection of TGF-beta1,α-SMA,Ⅰ,type Ⅲ collagen expression.Fully automatic image analysis system detects high content of paclitaxel inhibition concentration（IC50）48hours after the bile duct scar fibroblast cell cycle.[Results]1.The benign biliary scar into fiber cells that covered the bottom of culture flask,fusiform cells form a single,highly proliferative recovery two days.2.MTT assay showed that:the concentration of paclitaxel into benign biliary scar fibroblast drug intervention group and non-intervention group（negative control group）Inhibition of a statistically significant difference（p<0.05）;concentrations of paclitaxel on human benign bile duct scarof cell proliferation was significantly inhibited,the drug concentration-time-dependent inhibition;within a range of2.65625mg/ml～170mg/ml,the higher the concentration of paclitaxel on cell growth inhibition rate was higher（P<0.05）;drug action time is longer,the higher the inhibitory rate（P<0.05）.The 24h,48h,72h IC₅₀was 28.71mg/L,7.91mg/L,3.81mg/L,selected 48h IC₅₀concentration as a drug experiment.3.TUNEL method prompts benign biliary paclitaxel significantly promote apoptosis of scar fibroblasts.4.High content fully automatic image analysis system（HCS）and cell immunofluorescence method prompts alcohol into benign biliary scar fibroblast TGF-beta1,α-SMA,Ⅰand type Ⅲ collagen expression was significantly inhibited.5.High content automatic image analysis system（HCS）analysis showed that paclitaxel intervention bile duct scar fibroblasts arrest at the G₂phase and M phase.[Conclusions]Paclitaxel can significantly inhibit the benign biliary scar fibroblast proliferation,and has obvious drug concentration and time-dependent manner;promote the apoptosis cells were significantly arrested in the G₂and M phases;through inhibition of thecells TGF-β₁,α-SMA,Ⅰ and Ⅲ type collagen expression and to reduce the benign biliary scar formation.