title: Fasudil inhibits proliferation and collagen synthesis and induces apoptosis of human fibroblasts derived from urethral scar via the Rho/ROCK signaling pathway.
abstract: Fasudil has shown antifibrotic effects in various fibrotic diseases. However, its effects on human urethral fibroblasts are unknown. This study evaluated the effects of fasudil on cellular proliferation, migration, apoptosis, and collagen synthesis in human fibroblasts derived from urethral scar tissues. Human urethral scar fibroblasts were cultured by explant and incubated for 24 h or 48 h with fasudil (12.5, 25, 50 µmol/L) with or without transforming growth factor β1 (TGF-β1, 10 ng/mL), or left untreated (control). Cell proliferation and migration was determined by MTT assay and Transwell chambers, respectively. Apoptosis was measured by flow cytometry. Levels of α-smooth muscle actin (α-SMA), myosin light-chain phosphatase (MLCP), LIM domain kinase 1 (LIMK1), phospho-cofilin (p-cofilin), collagen I, and collagen III were determined by Western blot. Compared with the control group, TGF-β1 was associated with a significant increase in urethral fibroblast proliferation and migration, and α-SMA, MLCP, LIMK1, p-cofilin, collagen I, and collagen III levels. Compared with the control group, fasudil (with or without TGF-β1), significantly and negatively correlated, in a dose-dependent manner, with the proliferation and migration of urethral fibroblasts, as well as α-SMA, MLCP, LIMK1, p-cofilin, collagen I, and collagen III levels. Moreover, fasudil significantly induced apoptosis of fibroblasts induced by TGF-β1. Higher concentrations of fasudil (50 μmol/L) were associated with greater cell apoptosis without TGF-β1 stimulation compared with the normal control group. Fasudil, with or without TGF-β1 stimulation, may inhibit human urethral fibroblasts proliferation, migration, apoptosis, and collagen synthesis via the Rho/ROCK signaling pathway.
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Xu
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Shao-Hao
authors.initials: SH
authors.lastname: Chen
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Gen-Yi
authors.initials: GY
authors.lastname: Qu
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Xiao-Dong
authors.initials: XD
authors.lastname: Li
authors.affiliation: Department of Urology, Chinese PLA 476 Hospital Fuzhou, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Lin
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Xue-Yi
authors.initials: XY
authors.lastname: Xue
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Yun-Zhi
authors.initials: YZ
authors.lastname: Lin
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Qing-Shui
authors.initials: QS
authors.lastname: Zheng
authors.affiliation: Department of Urology, First Affiliated Hospital of Fujian Medical University Fuzhou, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wei
keywords: Fasudil; Rho kinase inhibitor; collagen synthesis; extracellular matrix; urethral fibroblasts