Effect of aprotinin on in vitro cerebral endothelial ICAM-1 expression induced by astrocyte-conditioned medium.
OBJECTIVE
Aprotinin administration may decrease the incidence of stroke associated with coronary artery bypass surgery by an unknown mechanism. Astrocytes exposed to hypoxia produce proinflammatory cytokines and upregulate intercellular adhesion molecule (ICAM)-1 on cerebral endothelium. This study investigated the effects of aprotinin on cerebral endothelial activation by hypoxic astrocytes in vitro.


METHODS
Mouse astrocytes were exposed to hypoxia in an anaerobic chamber for 4 h followed by reoxygenation for 24 h. Astrocyte-conditioned medium (ACM) collected from mouse astrocytes subjected to hypoxia/reoxygenation (HR) or normoxia were applied to mouse cerebral endothelial cell (MCEC) cultures for 4 and 24 h in normoxia. Endothelial cells were preincubated for 1 h with aprotinin (1600 KIU mL(-1)) prior to exposure to ACM. Flow cytometry was used to estimate endothelial ICAM-1 expression. Interleukin (IL)-1beta space concentrations in ACM were estimated with enzyme-linked immunosorbent assay (ELISA). Repeated comparisons were made using analysis of variance (ANOVA) and post hoc Tukey test as appropriate. P < 0.05 was considered significant. Data is presented as mean (standard deviation, SD).


RESULTS
MCEC ICAM-1 expression was greater after 24 h exposure to HR-ACM compared to normoxic-ACM (mean channel flouresence (MCF) 107.5 (4.5) vs. 74.3 (4.5), respectively, P < 0.001). ICAM-1 expression was decreased by aprotinin preincubation compared to control (MCF 91.0 (1.1) vs. 107.5 (2.1), P = 0.006). Supernatant IL-1beta concentrations in astrocytes exposed to HR were greater than those exposed to normoxia (7.1 (0.2) vs. 4.1 (0.2), P = 0.01).


CONCLUSIONS
This may be a neuroprotective mechanism associated with aprotinin administration.
Harmon
D
D
Walton Centre for Neurology and Neurosurgery, Liverpool, UK. dominicharmon@hotmail.com
Ghori
K
K
Lan
W
W
Shorten
G
G
D000693:Anaerobiosis; D000818:Animals; D007611:Aprotinin / Q000494:pharmacology*; D001253:Astrocytes / Q000502:physiology*; D001923:Brain Chemistry / Q000187:drug effects*; D015687:Cell Hypoxia / Q000502:physiology; D002460:Cell Line; D017077:Culture Media, Conditioned; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis*; D007375:Interleukin-1 / Q000378:metabolism; D051379:Mice; D018696:Neuroprotective Agents / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*
D017077:Culture Media, Conditioned; D007375:Interleukin-1; D018696:Neuroprotective Agents; D015842:Serine Proteinase Inhibitors; D018799:Intercellular Adhesion Molecule-1; D007611:Aprotinin