| ObjectiveTo investigate the effect of tranilast on proliferation of cultured humanretinal pigment epithelial (RPE) cells in vitro.Methods1. To treate them with different concentrations of tranilast0(controlgroup),12.5,25,50,100mg/l.after culturing human retinal pigment epithelial cells（hRPE-19cell line）3-5generations.And then to observe cell morphologyunder inverted microscope combined with HE staining,identify the cell bykeratin CK18.2. Detecting inhibition rate by MTT colorimetric method after the cellswere treated with different concentrations of tranilast for12h,24h,48h.3. The RPE cells were cultured with different concentrations of tranilast for24h,then to meature Transforming growth factor β1（TGF-β1）and growth factorreceptor A protein (PDGFR-A) expressing by Western Blot andimmunohistochemical.Results1. The hRPE-19cells were adhered to the wall and the cells morphologywere Irregular polygonal, cytoplasm contained a small amount of pigmentgranules. After processed with cytokeratin CK18the cytoplasmic was brownedpositive. 2. The MTT results showed cell inhibition rate increased with the increaseof concentration at the same time piont(p<0.05).Here was statisticallysignificant difference.3.The Western Blot results showed, the gray value of which bands waslowered with the increase of concentration of tranilast, and the TGF-beta1protein and PDGFR-A protein were lowered too(P＜0.05).Here was statisticallysignificant difference.4.TGF-beta1protein expression in cell plasma and PDGFR-A protein inthe cell membrane and cytoplasm,the expression of their amount was loweredwith the increase of concentration of tranilast (P＜0.05).Here was statisticallysignificant difference.ConclusionsTranilast could inhibit rpe proliferated, The cell inhibition rate increasedwith the increase of concentration at the same time piont(P＜0.05).Here wasstatistically significant difference.It may be connected with the TGF-β1andPDGFR-A protein expression decreased. |
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