YC-1 Reverses Retinal Reactive Gliosis and Retinal Neovascularization, <i>in vivo</i> and <i>in vitro</i>.

<p>Neovascular sprouts from DMSO-treated retinas had approximately similar lengths as the non-treated ischemic retinas [A]. YC-1 treated animals exhibited a significant and a dose-dependent decrease in neovascular sprouts lengths as compared to DMSO-treated retinas. [ANOVA, *<i>**P<0.001</i>; <i>**P<0.01</i> between YC-1 and DMSO] [A]. DMSO has no influence on the number of neovascular sprouts [B]. Whereas YC-1 exhibited a significant and a dose-dependent decrease in neovascular sprouts number [B]. [ANOVA, *<i>**P<0.001</i>; <i>**P<0.01</i> between YC-1 and DMSO]. There were 24 neovascular sprouts/retina that were averaged. [n = 5 per group]. By using primer sets [C]; Real Time RT-PCR analyses, <i>in vivo</i> [D and E] indicate that the levels of <i>GFAP</i> and <i>PDGF-B</i> mRNA were increased in the non-treated ischemic retinas, while non-treated normoxic retinas exhibited extremely low levels. Treatment of ischemic retinas with dual injections of YC-1 resulted in a significant knockdown of PDGF-B [**<i>P</i><0.01] and GFAP [**<i>P</i><0.01] gene expression when compared with DMSO-treated controls. ANOVA; Mean ± SEM of mRNA level normalized to β-actin were calculated, [***<i>P</i><0.001 and **<i>P</i><0.01, as compared to DMSO-treated retinas]. Data are representative of 3 independent experiments. Real Time RT-PCR analysis, <i>in vitro</i> [F] indicate that post-culturing R28 and rMC-1 cells under normoxic and hypoxic conditions; the mRNA levels of <i>GFAP and PDGF-B</i> were upregulated in all non-treated hypoxic cells, while normoxic cells exhibited remarkable low mRNA levels. Treatment of hypoxic R28 and rMC-1 cells with various concentrations of YC-1 resulted in a significant inhibition of <i>GFAP and PDGF-B</i>, mRNA expression as compared to DMSO-treated controls. ANOVA was used for statistical analyses. Mean ± SEM of mRNA level normalized to β-actin were calculated, [***<i>P</i><0.001 and **<i>P</i><0.01 as compared to DMSO-treated hypoxic control. Data are representative of 3 independent experiments].