The Role And Mechanism Of Fasudil On Urethral Scar Fibroblast Cell Extracellular Matrix Synthesis

ObjectiveThe study by Rho kinase inhibitor-fasudil under TGF-β1in vitro stimulation ofurethral fibroblasts proliferation, and extracellular matrix synthesis and involving inthe contraction of the urethral scar of cytoskeleton-associated proteins, such as α-actin(α-SMA), phosphorylated myosin light chain (myosin light chain phosphorylation,p-MLC), LIM kinase1(LIM kinase, LIMK-1) phosphorylated cofilin (Cofilinphosphorylation, p-Cofilin), and collagen I and III, of the effects, further reveal thepathogenesis of urethral scarring, provide new theories and methods for thepathogenesis of various fibrotic diseases.MethodsTake the urethra scar primary human fibroblasts were cultured. Divided intoeight experimental groups, the culture medium were added TGF-β1(10ng/ml), fasudil(12.5μmol/L,25μmol/L,50μmol/L), TGF-β1(10ng/ml)+fasudil (12.5μmol/L,25μmol/L,50μmol/L) and the control group. Detection of cell viability by MTT assay.with apoptosis rate of flow cytometry. Western-blot to detect the expression of α-SMA,p-MLC, LIMK-1, p-Cofilin, and I and III collagen cell viability.ResultsTGF-β1significantly increased viability of urethra fibroblasts, can induce theexpression of α-SMA p-MLC p-Cofilin, LIMK, I and III collagen, with theconcentration of which increases, induction of TGF-β1was gradually increasing trend.With increasing concentration of fasudil intervention, decreased activity of urethrafibroblasts; α-SMA, p-MLC, LIMK-1, p-Cofilin, and the expression of collagen I andIII decrease; Flow cytometry showed that in the absence of TGF-β1effect, higherconcentrations of fasudil group of early apoptosis rate was higher than control team (P <0.05); join under the action of TGF-β1, fasudil to urethra into the early apoptoticfibroblasts in a concentration-dependent manner.ConclusionsFasudil may inhibit TGF-β1induced by Rho/Rock signaling pathway in humanurethral fibroblast proliferation, migration, transformation, and synthesis ofextracellular matrix (ECM) and other biological behavior, and induce early apoptosis.Cytoskeleton-associated protein movement makes α-SMA, p-MLC, LIMK-1,p-Cofilin expression decreased, and a dose-dependent manner.