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Abstract #20596 Published in IGR 10-1
Effect of TGF-β1 on proliferation of Tenon capsule fibroblasts and expression of heat shock protein 47
Pan R; Chen X; Li M; Liu DChinese Ophthalmic Research 2007; 25: 933-936
OBJECTIVE: Glaucoma is a severe leading blindness disease. Glaucoma filtration surgery is the most popular strategy applied to reduce the intraocular pressure. However, the proliferation and collagen formation resulted from the fibroblasts lead to the obstruction of the created fistula and subconjunctival filtration area. This study aimed to investigate the effect of transforming growth factor beta 1 (TGF-β1) on the proliferation of human tenon capsule fibroblasts (HTFs) and expression of heat shock protein 47 (HSP47). METHODS: HTFs from glaucoma patients were cultured and passaged in DMEM medium supplemented with 10% fetal bovine serum. The 4 th and 6 th generations of HTFs were incubated with various concentrations of TGF-β1 (0.01, 0.1, 1, 5, 10 ng/mL) for 24 hours. The effects of TGF-β1 on the proliferation of HTFs was evaluated by MTT colorimetry. The expression of HSP47 and collagen type I protein was detected by immunocytochemistry, and the induction of HSP47 and collagen type I was evaluated at the mRNA level using quantitative real-time PCR (RT-PCR). RESULTS: Absorbent value of HTFs for different concentrations of TGF-β1 (0.01, 0.1, 1, 5, 10 ng/mL) was 0.237 &plusm; 0.004, 0.244 ± 0.010, 0.273 ± 0.012, 0.297 ± 0.011, 0.344 ± 0.012 respectively, showing that TGF-β1 could enhance the proliferation of HTFs. The integral optical density (IOD) value of HSP47 in TGF-β1-treated HTFs (0.01, 0.1, 1, 5, 10 ng/mL) was 23.65 ± 1.18, 24.13 &plusm; 1.15, 36.53 ± 1.18, 55.50 ± 1.25, 61.92 ± 1.17 respectively, and the that of collagen type I was 24.21 ± 1.24, 24.91 ± 1.72, 45.15 &plusm; 1.56, 60.57 ± 1.31, 74.79 ± 1.01 respectively, indicating that TGF-β1 could induce the expression of HSP47 and collagen type I. The mean values of relative expression showed 2.746-fold increase in HSP47 mRNA and 3.24-fold increase in collagen type ImRNA with 10 ng/mL TGF-β1 treatment. The expression of both HSP47 and collagen type I mRNA elevated with increasing TGF-β1 concentrations. CONCLUSION: TGF-β1 can promote the fibrosis of HTFs through enhancing the proliferation of HTFs and inducing the expression of HSP47 and collagen type I. It might be an important pathway in scarring of glaucoma after trabeculectomy. LA: Chinese
Dr. R. Pan, Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu 610041, China
Classification:
12.8.10 Woundhealing antifibrosis (Part of: 12 Surgical treatment > 12.8 Filtering surgery)
11.14 Investigational drugs; pharmacological experiments (Part of: 11 Medical treatment)
3.6 Cellular biology (Part of: 3 Laboratory methods)