Figure 6. Effects of FFA on the integrity of blood-brain barrier after cardiac arrest and cardiopulmonary resuscitation.
Representative photomicrographs of immunofluoresence staining with IgG antibody (red) in the cerebral cortex, striatum and hippocampus of the sham group and the experimental groups at 3 days after return of spontaneous circulation. Scale bar, 100 μm or 50 μm. (B ) Representative photomicrographs of immunofluorescence staining for tight junction proteins (green), including Occludin, Claudin-5, ZO-1, and CD31 (red) with DAPI (blue) in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation. Scale bars, 10 μm. (C ) Semi-quantitative results of IgG leakage. (D ) Quantitative analysis of Occludin/CD31 ratio, Claudin-5/CD31 ratio and ZO-1/CD31 ratio. ***P <0.001 versus the sham group; ##P <0.01 versus the vehicle group. FFA: flufenamic acid.Figure 7. Effects of FFA on microglia/macrophages polarization after cardiac arrest and cardiopulmonary resuscitation.
(A ) Representative photomicrographs of immunofluorescence staining for CD16/32 (green), Arg1 (green) and Iba1 (red) in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation. Scale bar, 50 μm. (B ) Quantification of the percentage of CD16/32 and Iba1 double-positive cells as well as Arg1 and Iba1 double-positive cells. (C ) qPCR results of pro-inflammatory and anti-inflammatory markers in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation. (D-E ) Western blot analysis of pro-inflammatory and anti-inflammatory markers in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation (D ) and quantitative graph (E ). (F ) Representative photomicrographs of immunofluorescence staining for Iba1 (green) and NeuN (red) in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation. Scale bar, 50 μm. (G ) Quantification of the number of Iba1 and NeuN double-positive cells. (H ) Representative photomicrographs of immunofluorescence staining for ICAM-1 in the brain of the sham group and the experimental groups at 3 days after return of spontaneous circulation. Scale bar, 100 μm. (I ) Semi-quantitative results of ICAM-1. *P <0.05, **P <0.01, ***P <0.001 versus the sham group; #P <0.05, ##P <0.01, ###P <0.001 versus the vehicle group. FFA: flufenamic acid; Iba1: ionized calcium-binding adapter molecule 1; ICAM-1: intercellular cell adhesion molecule-1. NeuN: neuronal nucle.