Promoting effect of connective tissue growth factor on expression of E-cadherin in human Tenon capsule fibroblasts / 中华实验眼科杂志

ABSTRACT

Background Scarring of filtration channel following glaucoma filtering surgery is a main cause of the failure of the surgery.The proliferation,epithelial-mesenchymal transition and extracellular matrix remodeling of fibroblasts are thought to be the primary pathological mechanism of scarring.Connective tissue growth factor (CTGF) plays a promoting role in the formation of scar.Whether CTGF participates in mesenchymal-epithelial transition of human Tenon capsule fibroblasts (HTFs) is not clear yet.Objective This study attempted to investigate the effect of CTGF on the mesenchymal-epithelial transition of HTFs in vitro.Methods HTFs were cultured and passaged in high glucose DMEM medium with 10％ fetal bovine serum,and the cells of generation 3-6 were used in this experiment.The cells were divided into the blank control group and CTGF-treated group and were routinely cultured in the blank control group.CTGF was added in the medium in the CTGF-treated group,with the final concentration 50 ng/ml CTGF.Immunofluorescence staining was used to identify and locate the expression of E-cadherin protein in the cells,and Western blot assay was employed to quantitatively analyze the expression level of E-cadherin protein in 48 hours after culture.Results The HTFs grew well with the spindle-like shape and vortex-like arrangement.The red fluorescence (E-cadherin protein) in the cytoplasm and blue fluorescence in the cellular nucleus were seen in the CTGF-treated group,but only nucleus with blue fluorescence were obtained in the blank control group.Western blot assay showed that the E-cadherin protein expression was absent in the blank control group,however,the relative expression level of E-cadherin protein in the cells was 0.63± 0.08.Conclusions E-cadherin protein is not expressed in fibroblast derived from mesenchymal tissue.However,CTGF can induce the expression of E-cadherin in HTFs.This study suggests that CTGF promotes the mesenchymal-epithelial transition of HTFs in vitro.