Attenuation of Choroidal Neovascularization by Histone Deacetylase Inhibitor
Fig 5
TSA reduces HIF-1α and VEGF expression and up-regulates the expression of PEDF.
Real-time PCR (A) and Western blot assays (B) were performed on RPE cells treated with 0–0.5 μM TSA and 150 μM CoCl2. (A) Cells were treated with 0–0.5 μM TSA for 14 h and then co-treated with 150 μM CoCl2 for 6 h for the analysis of gene expression by real-time PCR. Changes in HIF-1α mRNA levels were not statistically significant. CoCl2 causes a fourfold enhancement of VEGF mRNA expression; but at 0.5 μM TSA, the mRNA level of VEGF reduces to less than half of that in cells treated with CoCl2 only. TSA induces a statistically significant increase in the mRNA level of PEDF. (B) Cells were treated with 0–0.5 μM TSA for 18 h and then co-treated with 150 μM CoCl2 for 6 h for Western blot analysis. TSA reduces the CoCl2-induced HIF-1α and VEGF protein levels by 4.3-fold and 5.7-fold, respectively, and up-regulates PEDF protein level by threefold. (C) Densitometry data for Western blot of HIF-1α, VEGF and PEDF. (*: t test p<0.05; **: t test p<0.01).