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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.
This study attempted to investigate the effect of CTGF on the mesenchymal-epithelial transition of HTFs in vitro.
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.
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.
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.