Figure 8

Effect of intravitreal fasudil on GK rat retinal vessel dilation. (A) Confocal in vivo fluorescein angiography before and after fasudil treatment allowed visualization of retinal vessels (a–d). Edge detection was performed using a dedicated macro (b–e,c–f) and vascular diameter before and after treatment determined a vasodilation ratio (g) showing significant effect of fasudil on retinal vessel diameter (p < 0.05). (B) An automated detection of retinal capillary network surface was performed on retinal flat-mounts stained by TRITC-lectin (red) to highlight the vessel walls in vehicle and Fasudil treated GK rats (small vessels represented in blue and large vessels in red) (a–b and c–d). The retinal surface coverage by small vessels was significantly increased in fasudil treated GK rats as compared to vehicle treated GK rats (n = 4–5 per group, p < 0.05) consistent with a vasodilation of retinal capillaries (e). (C) Western blot showed a significant decrease in the VEGF-164 level in the neural retina of fasudil treated GK rats as compared to vehicle treated GK rats, suggesting decreased retinal hypoxia in fasudil treated retina (n = 8 per group). *p value < 0, 05; **p value < 0, 01 ***p value < 0,001.