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.