Antagonistic Effects of Tranilast on Proliferation and Collagen Synthesis Induced by TGF-β_2 in Cultured Human Trabecular Meshwork Cells

摘要

Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-β_2 in cultured human trabecular meshwork cells was investigated. Suspension of 1X10~4 cultured human trabecular meshwork cells of 3—5 passage was distributed in each well of a 96-well disk and divided into control group and experimental group. After 24 h, 0 μg/ml (control), 12. 5 μg/ml, 25 μg/ml, 50 μg/ml tranilast with 3. 2 ng/ml TGF-β_2 were added into the incubation medium. Another 24 h later, proliferation and collagen synthesis in cultured human trabecular meshwork cells were examined respectively by using tetrazolium-based semiautomated color-metric (MTT) assay and ~3H-proline incorporation with liquid scintillation technique. The results showed absorbance (A) values of the experimental groups were 0. 9036 +- 0. 3017, 1. 1361 +-0.1352, 1. 2457 +- 0. 1524 according to the different concentrations of tranilast, and 0. 8956 +-0. 1903 of the control group. In comparison with the control group, 25 μg/ml (q′=3. 23, P < 0. 05) , 50 μg/ml (q′ = 4. 70, P < 0. 01) tranilast significantly antagonized the decrease of the A values induced by TGF-β_2 in the cultured human trabecular meshwork cells. In. comparison with the control group [817. 37+-124. 21 cpm/10~4 cells], 12. 5 μg/ml (620. 33+-80. 46 cpm/10~4 cells, q′= 4.26, P < 0.05), 25 μg/ml (594. 58 +- 88. 13 cpm/10~4 cells, q′=4. 81, P < 0. 01), 50 μg/ml (418. 64+-67. 90 cpm/10~4 cells, q′ = 8. 62, P < 0. 01) tranilast significantly inhibited the incorporation of ~3H-proline into the cultured human trabecular meshwork cells promoted by TGF-β_2 in a dose-dependent manner. It was concluded that tranilast had the antagonistic effect on the proliferation inhibition and collagen synthesis promotion induced by TGF-β_2 in the cultured human trabecular meshwork cells.