Effect Of Apigenin On Fibroblasts Of Benign Bile Duct Scar In Human

Objectives:Celery element on the original generation of in vitro cultured human benign bile duct scar fibroblasts proliferation,migration,apoptosis,cell cycle,and observation of cell TGF-?1,?-SMA,Collagen ? and Collagen ? protein expression level,explore the celery,the application prospect in the treatment of benign bile duct stricture and the possible mechanism.Methods:In vitro primary culture of benign bile duct scar fibroblasts were identified,and the fourth or fifth generation cells were randomly divided into the experimental group and the control group.OD values of apigenin at different concentrations(80?mol/L,40?mol/L,20?mol/L,10?mol/L,5?mol/L)were detected by CCK-8 method,and the semi-inhibitory concentration(IC50)values were calc?lated.CCK-8 and EDU-488 methods were used to observe the effect of apigenin on cell proliferation indirectly and directly.Cell scratch and Transwell were used to detect the effect of apigenin on cell migration.Annexin V and PI were used to label cells and flow cytometry was used to detect apigenin-induced apoptosis.The cells were labeled by PI and the cell cycle of apigenin was detected by flow cytometry Using cell immunofluorescence and Western Bolt testing celery element after the intervention cells TGF-?1,?-SMA,Collagen ? and Collagen ? protein expression All the study data were statistically analyzed by SPSS 21.0.Res?lts:The primary culture cells grew logarithmically,and the cells were in the shape of mononuclear shuttle.Vimentin and Keratin were used to test the strongly positive expression(Green fluorescence)of the cells to prove that this cell is HBBDSF.The CCK-8 method shows that the calculated result of selecting OD value data of 48h is taken as the IC50 value of apenin,IC50=22.75 mol/L,R2=0.797.At the observation time points of 24h,48h and 72h,compared with the control group,apigenin treatment group had a significant inhibitory effect on the proliferation of HBBDSF(P<0.05),and had a concentration dependence.EDU-488 method showed that compared with the control group,apigenin treatment group had a significant inhibitory effect on the proliferation of HBBDSF(P<0.05),and had a concentration dependence.Both cell scratch and Transwell(Migration experiment)showed that compared with the control group,apigenin treatment group had a significant inhibitory effect on the migration ability of HBBDSF(P<0.05)and had a concentration dependence.Annexin V and PI double standard methods showed that compared with the control group,apigenin treatment group significantly promoted the apoptosis of HBBDSF(P<0.05),and the concentration was dependent.Detection of apigenin's effect on cell cycle by PI single standard method showed that compared with the control group,apigenin treatment group could significantly inhibit the G1/S phase of HBBDSF(P<0.05),and it was concentration-dependent.Cell immunofluorescence and Western Bolt testing celery element after the intervention cells TGF-?1,?-SMA,Collagen ? and Collagen ? protein expression of the situation,found that compared with the control group had obvious inhibitory effect(P<0.05),and has the concentration dependenceConclusion(s):1.Successfully established the experimental model of human benign bile duct scar fibroblasts;2.Apigenin can inhibit the proliferation of HBBDSF by blocking their G1/S phase;3.Apigenin can inhibit the migration of HBBDSF by inhibiting TGF-?1 pathway;4.Apigenin can induce the apoptosis of HBBDSF;5.Celery element after intervention in HBBDSF,the cells of TGF-?1,?-SMA,Collagen ? and Collagen ? protein expression level decreased obviously.