Figure 6
The effect of A-779 on the clinical scores, retinal mRNA expression of the cytokines and the surface markers of M1/M2 macrophages.
(a) Clinical signs were assessed with a slit lamp from day 7 to day 21 after the mice received IRBP immunization and the implantation of mini-pumps with A-779 or vehicle. Representative images showed the anterior inflammation from the AAV8-ACE2 or AAV8-eGFP treated EAU mice and AAV8-ACE2 treated EAU mice with the A-779 or vehicle implantation at the 14th day after immunization. (b) The severity of clinical scores were remarkably decreased on the 12th, 14th day in the AAV8-ACE2 treated eyes compared with the AAV8-eGFP treated EAU eyes (***P < 0.001) and AAV8-ACE2+A-779+EAU eyes (##P < 0.01, ###P < 0.001, n = 4–8 per group). The retinas were dissected at the 14th day after IRBP immunization for analyzing the mRNA levels of the pro-inflammatory and anti-inflammatory cytokines and the surface markers of M1/M2 macrophages. (c) The mRNA levels of pro-inflammatory cytokines: IL-6, IL-1β, TNF-α and MCP-1 and anti-inflammatory cytokine: IL-10 in the AAV8-ACE2+EAU, AAV8-eGFP+EAU, AAV8-ACE2+EAU+A-779 and AAV8-ACE2+vehicle+EAU groups. (d) The mRNA expression of Th1 cell signature cytokine IFN-γ and Th17 cell signature cytokine IL-17 were remarkably reduced in the AAV8-ACE2 treated EAU eyes compared with the AAV8-eGFP treated eyes. A-779 abrogated the reduction of IFN-γ and IL-17. (e) The mRNA of iNOS was decreased, whereas the mRNA of Arg-1 was increased in the AAV8-ACE2+EAU group. However, A-779 reversed the effect of ACE2 on the polarization of M1/M2 macrophages. Data were expressed as mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001, n = 4–6 per group).
