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BACKGROUND: Researches indicate that transforming growth factor-β-2 (TGF-β-2) plays an important role in the cicatrisation of filtering bleb after glaucoma filtering surgery. OBJECTIVE: To observe the influence of anti-transforming growth factor-β-2 on the proliferation of filtering bleb fibroblasts cultured in vitro and its expression of p21 and cyclinD1. DESIGN, TIME, SETTING: The open experiment was carried out in June 2007 at the Department of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China. MATERIAL: Scar tissues removed from failed filtering blebs after glaucoma trabeculectomy. All patients were male and their age ranged from 23 to 35 years; eight patients did not receive anti-scar drug treatment before and during the operation. METHODS: Primary cell culture was done using the scar tissue to get fibroblasts. Fibroblasts were incubated with 0 (control), 0.01, 0.1, 1, 10 mg/L of anti-TGF-β-2 antibody for 24 hours; inhibitory concentration 50 of anti-TGF-β-2 antibody was used to treat the fibroblasts for 0 (control), 24, 48, 72 hours, respectively. MAIN OUTCOME MEASURES: The influence of anti-TGF-β-2 antibody on the proliferation of fibroblasts and its expression of p21 and cyclinD1 observed with MTT and immunohistochemistry. RESULTS: With the increase of the anti-TGF-β-2 antibody concentration and the prolongation of the action time, the inhibitory effect of antibody on the fibroblasts became more and more obvious; the cyclinD1 positive expression decreased gradually while the weak positive expression increased; on the other hand, the strong positive expression of p21 raised noticeably and the expression in cell nucleus also increased (P < 0.01). The expression of p21 showed a marked negative correlation with the expression of cylinD1 under different concentrations of anti-TGF-β-2 antibody and at different time points (r = -0.91, r = -0.85, P < 0.05), p21 and cylinD1 both were correlated noticeably with the proliferation inhibition of cells (r = 0.76, P < 0.05). CONCLUSION: The anti-TGF-β-2 antibody can inhibit the proliferation of fibroblasts by increasing the synthesis of p21 protein and regulating the expression of cyclinD1. LA: Chinese
Dr. H.-L. Liao, Department of Ophthalmology, Hospital of Guangdong Medical College, Zhanjiang 524001 Guangdong Province, China. liaohailan@hotmail.com