Effect Of Tacrolimus On Transdifferentiation Induced By Tgf-β And Signal Transduction Pathway In Human Tenon's Fibroblasts

Scarring formationg of the filtering path leads to surgical failure after glaucoma filtering surgery. Now this situation is improved to some extent with surgery and anti-metabolic drugs use, but brought many complications still. Therefore,the search for inhibitior of scar formation ,which is both more effective and fewer side effects, has the great significance. Tacrolimus is one kind of widely used immunosuppressive agents,topical anti-rejection of corneal transplantation. Tacrolimus has been used in anti-fibrosis and inhitition excessive of wound healing research,but the effect of tacrolimus on human Tenon's capsule fibroblasts is unclear.In this study, human Tenon's capsule fibroblasts were cultured in vitro and induced by transforming growth factor-β,which transdifferentiation to myofibroblasts. Detection MF cells maker proteinα-smooth muscle actin(α-SM-actin) to identified the transdifferentiation. Different concentrations of tacrolimus were added into the transdifferentiated cells with 10, 20, 40, 80, 100nmol/l seperatedly. CCK8 assay was used to evaluate the vital force of HTF after measured 72h. Western-blot detection of cellα-SM-actin, connective tissue growth factor (CTGF), Collagen fiberⅠ(ColⅠ)expression. Western blot detection of Smad2, extracellular signal-regulated kinase(ERKs), p38 protein expression. The results showed that HTF cultured successfully in vitro, identified by immunocytochemistry confirmed. After TGF-β1 simulation with concertration of 1~40ng/ml, the protein expression ofα-SM-actin were up-regulated, to hint the HTF transdiffrentiation into MF. Inducting of 48h,α-SM-actin expression is significantly increasing trend. Compared with the control group,10~80nmol/l tacrolimus inhitit the proliferation of HTF which induced by TGF-β; 20~80nmol/l tacrolimus inhibitedα-SM-actin, CTGF and ColⅠprotein. Tacrolimus inhibits TGF-β-induced p-Smad expression, ERK and p38 signaling pathway has no effect. The effect of HTF transdifferentiation by tacrolimus through p-Smad pathway mainly.The study suggests that, TGF-βcan induce HTF transdifferent into MF. Tacrolimus inhibits TGF-βinduced transdifferentiation and dose-dependent manner. Tacrolimus activates the Smad signaling pathway to inhibit HTF transdifferentiation.