Figure 6

Exposure to hyperglycemic conditions, cigarette smoke extract and both combined promotes the pro-inflammatory and anti-oxidant defense activation in HCMEC/D3 BBB endothelial cells. a1 PECAM-1 expression in membrane fraction was significantly increased only in endothelial cultures following 12 h exposure to HG and HG + CSE. The magnitude of the effect significantly increased at 24 h exposure showing a consistent additive pattern between tested conditions (b1). a2 12 h HG, CSE and HG + CSE triggered upregulation of ICAM-1 membrane fraction. By contrast, although statistically significant, the upregulation of ICAM-1 did not show a longitudinal response pattern to the tested conditions (b2). c1, c2 Release of IL-6 modestly increased in HG-CSE treated cultures while an additive and more robust increased in IL-8 release was observed under the same conditions at 24 h. d Representative Western blot showing the translocation of Nrf-2 from cytosol to nucleus of HCMEC/D3 cells in response to the above mentioned tested conditions. The graph bars clearly show the nuclear vs. cytosol ratios of Nrf-2 under the tested conditions. n = 3–4 biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. control; ^$ p < 0.05, ^$$ p < 0.01, ^$$$$ p < 0.0001 vs. HG and ^# p < 0.05, ^#### p < 0.0001 vs. CSE.