Figure 1

Effect of mouse cytokines on bend-3 mono-culture barrier and bEnd-3/HFA co-culture barrier. a) Cumulative effect of mouse cytokines (TNF-α (20 ng/ml), IL-1β (20 ng/ml) and IFN-γ (1000 U/ml)) applied to apical + basal sides of mouse brain endothelial mono-cultures. Resistance was recorded daily (7 d). Significant increase in the resistance of mouse brain endothelium was observed in a rank order of IFN-γ > TNF-α > IL-1β compared with control. Inset shows the mode of culture and cytokine treatment. Bars indicate standard error. Repeated measured ANOVA with Dunnett's post-hoc test. *p < 0.05 was considered to be statistically significant, **p < 0.01 very significant, and ***p < 0.001 highly significant. b) Effect of mouse cytokines (TNF-α (20 ng/ml), IL-1β (20 ng/ml) and IFN-γ (1000 U/ml)) of mouse brain endothelial solute permeability. Solute permeability was measured at 30', 1 h, 2 h, 3 h, 4 h and 6 h after 3 d of treatment. TNF-α and IFN-γ treated cultures showed lesser permeability than control or IL-1b treated cultures. The solute permeability of mouse brain endothelium in this experiment was in a rank order of IFN-γ ≈ TNF-α > IL-1β ≈ Con. Bars indicate standard error. Repeated measured ANOVA with Dunnett's post-hoc test. *p < 0.05 was considered to be statistically significant, **p < 0.01 very significant, and ***p < 0.001 highly significant. c) Effect of mouse cytokines (TNF-α (20 ng/ml), IL-1β (20 ng/ml) and IFN-γ (1000 U/ml)) on contact dependent bEnd-3/HFA co-culture system. Resistance was recorded daily. Significant increase in mouse brain endothelial barrier was observed with IFN-γ > IL-1β ≥ TNF-α compared to controls. Inset shows the mode of contact dependent system used and cytokine addition. Bars indicate standard error. Repeated measures ANOVA with Dunnett's post-hoc test. d) Effect of mouse cytokines (TNF-α (20 ng/ml), IL-1β (20 ng/ml) and IFN-γ (1000 U/ml)) on contact independent bEnd-3/HFA co-culture system. Resistance was recorded daily. Significant increase in the resistance of brain endothelium was observed with IFN-γ > IL-1β ≥ TNF-α compared with control. Inset shows the mode of contact dependent system used and cytokine addition. Bars indicate standard error. Repeated measures ANOVA with Dunnett's post-hoc test. *p < 0.05 was considered to be statistically significant, **p < 0.01 very significant, and ***p < 0.001 highly significant.