Chemical hypoxia-ischemia induces apoptosis in cerebromicrovascular endothelial cells.

Cerebral endothelial cell (CEC) death from ischemia may exacerbate brain injury by altering microvascular integrity, but little is known concerning the pattern of CEC death and disruption of tight junction between two CECs to ischemia. To address these questions, CECs were isolated from bovine, cultured, and placed in glucose-free medium containing sodium cyanide. Trypan blue staining shown that sodium cyanide resulted in a dose-dependent insult of CECs (10-80 mM). CEC injury increased progressively with the duration of 20 mM cyanide exposure, becoming significant (71%) after 12 h. The mode of cell death induced by cyanide is clearly apoptosis in CECs, as shown by Hoechst 33,342 staining and transmission electron microscope, i.e. cyanide induced condensation and margination of chromatin, nuclear fragmentation and shrinkage of cell body and condensed apoptotic bodies in CECs. Most importantly, we found that the tight junction between two CECs was disrupted 12 h after chemical-ischemia, i.e. when CECs underwent apoptosis, the tight junctional complexes became thinner and rough; the cleft of tight junction between two CECs became blurred and more wider, and membranes of tight junction were course and irregular; and the adherens junctions were damaged. These results indicate that chemical hypoxia-ischemia induces apoptotic cell death in CECs and alters the microvascular integrity by disrupting tight junction complexes, and suggest that CEC apoptotic death and disruption of tight junction may exacerbate ischemic insults to brain. Thus, prevention of CEC apoptotic death may contribute to improvements of ischemic insults.