Figure 5.

AZD0530 treatment impedes the generation of new functional vessels in the penumbral regions of EC-targeted miR-15a/16-1 cKO mice after cerebral ischemia. EC-miR-15a/16-1 cKO and WT littermate controls were subjected to 1 h MCAO followed by 28d reperfusion. Vehicle (Veh) or AZD0530 (AZD, 20 mg/kg) was administered daily to both genotypes at 3-21d after MCAO by oral gavage. Tomato lectin (green) and BrdU (red) double-immunofluorescent staining was utilized to detect functional microvessels in the penumbral regions. The lectin⁺/BrdU⁺ signals exhibit a yellow color, and the yellow boxes indicated areas in (a) were enlarged and 3 D reconstructed in (b). (a–f) Representative images (a), representative 3 D reconstructed images (b), quantitative analysis of surface area (c), vascular length (d), branch points (e), and capillary number (f) of tomato lectin immunofluorescent signal showed more functional vessels in penumbral regions of EC-miR-15a/16-1 cKO mice than WT controls at 28 d reperfusion after MCAO, whereas AZD0530 treatment reduced the upregulated-functional vessels in EC-miR-15a/16-1 cKO mice after cerebral ischemia. (a,b,g,h) Representative tomato lectin and BrdU double-immunofluorescent images (a), representative 3 D reconstructed images (b), quantification of the BrdU⁺ cells (g) and lectin⁺/BrdU⁺ signals (h) in the brain penumbral regions indicate more newly generated functional vessels in the EC-miR-15a/16-1 cKO mice than WT controls, while AZD0530 treatment significantly impeded the generation of new functional vessels in the EC-miR-15a/16-1 cKO mice after cerebral ischemia. Data are expressed as the mean ± SD. n = 5-7 for each group. ^&p < 0.05 between MCAO WT + Veh and MCAO WT + AZD groups; *p < 0.05, **p < 0.01 between MCAO cKO + Veh and MCAO cKO + AZD groups; ^#p < 0.05 as indicated. Statistical analysis was performed by one-way ANOVA followed by Bonferroni's multiple comparison tests (Figure 5(c) to (f) and (h)) or Kruskal-Wallis test followed by Dunn's multiple comparison tests (Figure 5(g)).