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. 2007 Feb 22;580(Pt 3):937–949. doi: 10.1113/jphysiol.2007.129007

Figure 6. Glucocorticoid preservation of endothelial barrier function under pro-inflammatory conditions.

Figure 6

A, glucocorticoid treatment prevents a compromise of BBB function in response to TNFα administration: 8 h TNFα treatment led to a reduction in TER to 65 ± 8% of control values, while treatment with hydrocortisone or dexamethasone increased TER values to 160 ± 7% and 163 ± 6% of control values, respectively. Simultaneous administration of TNFα with hydrocortisone effectively prevented barrier breakdown, TER values amounted to 96 ± 7% of control values, while simultaneous administration of TNFα with dexamethasone even led to increased tightness of the barrier (TER = 145 ± 9% of control values). B, confluent monolayers of cEND cells were grown in gelatine-coated cell culture flasks to confluence for 5 days and maintained in differentiation medium containing 2% FCS for an additional 3 days. On day 3, cells were treated with 10 nm TNFα for 8 h and cell lysates prepared. Cell lysates were analysed by Western blot for occludin, claudin-5 and ZO-1. Eight hours of TNFα treatment caused a decrease in occludin protein of 21 ± 3% as compared to the untreated control. Simultaneous administration of TNFα and hydrocortisone or dexamethasone prevented occludin loss; we were even able to detect an increase in 17 ± 5% and 49 ± 7%, respectively, as compared to untreated cells. When treated with hydrocortisone and dexamethasone alone, occludin contents were increased by 22 ± 2% and 57 ± 4%, respectively. Eight hours of TNFα treatment caused a decrease in claudin-5 protein of 24 ± 4% as compared to the untreated control. Simultaneous administration of TNFα and hydrocortisone or dexamethasone, however, did not significantly prevent claudin-5 loss, claudin-5 protein levels were reduced by 26 ± 5% and 13 ± 3%, respectively, as compared to untreated cells. When treated with hydrocortisone and dexamethasone alone, claudin-5 contents did not change significantly (88 ± 7% and 93 ± 4%, respectively). Protein levels in ZO-1 were not changed significantly by either treatment.