Expression Of Thymosin Beta4 In Human Tenon's Capsule Fibroblasts And The Effect Of CTGF On Its Expression

Purpose:Thymosin beta4 (Tβ₄) is a main actin-sequestrating peptide that modulates inflammation and healing in different tissues. The purpose of this study was to investigate the molecular and biochemical expression of Tβ₄ in human Tenon's capsule fibroblasts(HTFs) and to investigate the effect of connective tissue growth factor (CTGF) on the expression of Tβ₄.Methods:Total RNA from cultured 3-4 passage HTFs in vitro was extracted with TRIZOL reagent. Tβ₄ messenger RNA was measured by reverse transcriptase-polymerase chain reaction (RT-PCR). An immunohistochemical stain was used to detect Tβ₄ protein. After treated with 0 ng/ml(control), 1.0, 10.0 and 100.0ng/ml CTGF for 24 h,Tβ₄ expression in cultured 3-4 passage human Tenon's capsule fibroblasts was measured by semi-quantitative RT-PCR .Results: A single RT-PCR amplified product was obtained. Tβ₄ immunohistochemical staining was positive in cytoplasm of HTFs, while negative in nucleus. The result of the semi-quantitative RT-PCR was analyzed by image analysis. Respectively, treated with1.0, 10. 0 and 100.0ng/ml CTGF, the values of Tβ₄/β-actin were 1.11±0.04,1.05±0.03,0.82±0.06. The difference between these treated groups and the control group (1.23±0.02) was statistically significant ( P < 0.05).Conclusion: The date provides evidence that Tβ₄ expression in HTFs and CTGF can decreased the expression of Tβ₄ in dose-dependent manner.This may be a possible mechanism in the bleb scarring formation process after glaucoma filtration surgery。...