The Function And Possible Mechanisms Of ADSCs To The Benign Bile Duct Scar Fibroblast On TGF-β/smad Signal Pathway

Objective:To study the function and possible mechanisms of transforming growth factor(TGF-β1) and adipose tissue-derived mesenchymal stem cells (ADSCs) in the benign bile duct scar fibroblast.Methods: 1. The resuscitation and serial subcultivation of adipose tissue-derived mesenchymal stem cells.2. The resuscitation and serial subcultivation of human benign bile duct scar fibroblast.3.Different treatment of human benign bile duct scar fibroblast,then grouping them:A. (blank group):human benign bile duct scar fibroblast.B.(ADSCs-interfered group);:human benign bile duct scar fibroblast+ADSCs (induced on the 7th day); C.(TGF-β1-stimulated group):human benign bile duct scar fibroblast+25ng/ml transforming growth factor; D.(adipose interfered TGF-β1-stimulated group):human benign bile duct scar fibroblast+25ng/ml transforming growth factor+ADSCs (induced on the 7th day)。4. Co-culturing adipose tissue-derived mesenchymal stem cells which is induced on the 7th day and human benign bile duct scar fibroblast. 5. Observating the growth condition of ABCD group of human benign bile duct scar fibroblast. Detecting the expression of alpha-smooth muscle actin(a-SMA) the collagen of type I and type III on human benign bile duct scar fibroblast by immunofluorescence technique, detecting the mRNA expression ofα-SMA,the collagen of type I and type III on human benign bile duct scar fibroblast by reversed transcription-polymerase chain reaction (RT-PCR).Results:1.Human benign bile duct scar fibroblast trained in vitro could express a-SMA, the collagen of type I and type III. 2. The result of MTT show:growth condition of human benign bile duct scar fibroblast treated by transforming growth factor (C group)is better than A group. The result of RT-PCR show:the mRNA expression of the a-SMA,collagen of type I and type III, smad2/smad3/smad4 is higher than A group,but smad7 do not have evident difference.It cue that there is TGF-β/smad Signal pathway on the benign bile duct scar fibroblast, TGF-β1 could activate TGF-β/smad Signal pathway on the benign bile duct scar fibroblast, facilitating the growth of benign bile duct scar fibroblast.3. The result of MTT show:growth condition of human benign bile duct scar fibroblast treated by ADSCs (B and D group)is poorer than A group.The result of RT-PCR show:the mRNA expression of the a-SMA,collagen of type I and type III, smad2/smad3/smad4 is lower than A group,but smad7 do not have evident difference. ADSCs induced on the 7th day could inhibit the growth of benign bile duct scar fibroblast, possibly by inhibiting the TGFβ/Smad pathway.Conclusions:1.Human benign bile duct scar fibroblast trained in vitro could express a-SMA, the collagen of type I and type III.2.TGF-β1 could activate TGF-β/smad Signal pathway on the benign bile duct scar fibroblast, facilitating the growth of benign bile duct scar fibroblast. Enhancing the mRNA expression of the a-SMA,collagen of type I and type III, smad2/smad3/ smad4.3. ADSCs induced on the 7th day could inhibit the benign bile duct scar fibroblast growth, inhibiting the mRNA expression of the a-SMA,collagen of type I and type III, smad2/smad3/smad4.4. ADSCs induced on the 7th day may inhibit the benign bile duct scar fibroblast growth by inhibiting the TGFβ/Smad pathway.