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Effect of midazolam on in vitro cerebral endothelial ICAM-1 expression induced by astrocyte-conditioned medium

  • K. Ghori (a1), D. Harmon (a1), F. Walsh (a1) and G. Shorten (a1)



Background and objective: Astrocytes exposed to hypoxia produce pro-inflammatory cytokines and upregulate intercellular adhesion molecule-1 on cerebral endothelium. This study investigated the effects of midazolam on this response. Methods: Mouse astrocytes were exposed to 4 h of hypoxia and 24 h of reoxygenation. Astrocyte-conditioned medium were applied to mouse cerebral endothelial cell cultures for 4 h and 24 h in normoxia. Endothelial cells were pre-incubated for 1 h with midazolam (0, 5, 50 μg L−1). Flow cytometry was used to estimate endothelial ICAM-1 expression. IL-1β concentrations were measured with ELISA. Repeated comparisons were made using ANOVA and post hoc Tukey Test as appropriate. Data are mean (SD). Results: Mouse cerebral endothelial cell ICAM-1 expression was greater after 24 h exposure to hypoxia–reoxygenation astrocyte-conditioned medium compared to normoxic astrocyte-conditioned medium (mean channel flouresence 112.5 (9.5) vs. 81.5 (7.5), P = 0.01). ICAM-1 expression was decreased by midazolam (5 μg L−1) compared to control (mean channel flouresence 81.35 (7.5) vs. 112.5 (9.5), P = 0.01). Supernatant IL-1β concentrations (pg mL−1) in astrocytes exposed to hypoxia–reoxygenation were greater than those exposed to normoxia (16.41 (2.35) vs. 10.5 (2.13), P = 0.01). Conclusion: We conclude that decreased cerebral endothelial ICAM-1 expression in response to activated glial cell compartment by midazolam may decrease post ischaemic brain inflammation and secondary brain injury.


Corresponding author

Correspondence to: Kamran Ghori, Department of Anaesthesia and Intensive Care Medicine, Cork University Hospital, Wilton Cork, Ireland. E-mail:; Tel: +353 214 546 400; Fax: +353 214 546 434


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Jean WC, Spelman SR, Nussbaum ES, Low WC. Reperfusion injury after focal cerebral ischemia: the role of inflammation and the therapeutic horizon. Neurosurgery 1998; 43: 13821396.
Del Zoppo GJ. Microvascular changes during cerebral ischemic and reperfusion. Cerebrovasc Brain Metab Rev 1994; 6: 4796.
Del Zoppo GJ, Wanger S, Tagaya M. Trend and future developments in the pharmacological treatment of acute ischemic stroke. Drug 1997; 54: 938.
Feuerstein GZ, Liu T, Barone FC. The role of cytokines in the neuropathology of stroke and neurotrauma. Neuroimmunomodulation 1998; 5: 143159.
Feuerstein GZ, Liu T, Barone FC. Cytokine, inflammation, and brain injury: role of tumor necrosis factor α. Cerebrovasc Brain Metab Rev 1994; 6: 341360.
Neary P, Redmond HP. Ischemia-reperfusion injury and the systemic inflammatory response syndrome. In: Grace PA, Mathie RT, eds. Ischemia-Reperfusion Injury.London: Blackwell Science, 1999: 123136.
Barone FC, Hillegass LM, Price WJet al. Polymorphonuclear leukocyte infiltration into cerebral focal ischemic tissues: myeloperoxidase activity assay and histologic verification. J Neurosci Res 1991; 29: 336348.
Lindsberg PJ, Carpen O, Paetau Aet al. Endothelial ICAM-1 expression associated with inflammatory cell response in human ischemic stroke. Circulation 1996; 94: 939945.
Prober JS. Cytokine mediated activation of vascular endothelium: physiology and pathology. Am J Pathol 1988; 133: 426433.
Giulian D. Microglia, cytokine and cytotoxins: modulators of cellular responses after injury to the central nervous system. J Immunol Immunopharmacol 1990; 10: 1521.
Stanimirovic D, Satoh K. Inflammatory mediators of cerebral endothelium: A role in ischemic brain inflammation. Brain Pathol 2000; 10: 113126.
Zhang W, Smith C, Howlett C, Stanimirovic D. Inflammatory activation of human brain endothelial cells by hypoxic astrocytes in-vitro is mediated by IL1β. J Cereb Blood Flow Metabolism 2000; 20: 967978.
Zhang W, Smith C, Shapiro Aet al. Increased expression of bioactive chemokines in human cerbromicrovascular endothelial cells and astrocytes subjected to simulated ischemia in vitro. J Neuroimmunol 1999; 101: 148160.
Bradbury MB. The blood–brain barrier. Exp Physiol 1993; 78: 453472.
Ito H, Watanabe Y, Isshiki A, Uchino H. Neuroprotective properties of propofol and midazolam but not pentobarbital, neuronal damage induced by forebrain ischemia, based on the GABAA receptors. Acta Anaesthesiol Scand 1999; 43 (2): 153161.
Kirsch J, Traystman R, Hurn P. Anesthetics and cerebroprotection: experimental aspects. Int Anesthesiol Clin 1996; 34: 7393.
Nugent M, Artru A, Michenfelder D. Cerebral metabolic, vascular and protective effect of midazolam maleate: comparison to diazepam. Anesthesiology 1982; 56: 172176.
Kellbel I, Weiss M. Anaesthetics and immune function. Curr Opin Anesthesiol 2000; 14: 685691.
Kahoru N, Hirohiko A, Katssuya Met al. The Inhibitory effect of thiopentone, midazolam and ketamine on human neutrophil function. Anesthesiology 1998; 86: 159165.
Chen L, Gong Q, Xiao C. Effects of propofol, midazolam and thiopental sodium on outcome and amino acids accumulation in focal cerebral ischemia-reperfusion in rats. Chin Med J (Engl) 2003; 116: 292296.
Abbott J, Revest A, Romero AA. Astrocyte–endothelial interactions; physiology and pathology. Neuropathol Appl Neurobiol 1992; 18: 424433.
Winquist J, Kerr S. Cerebral ischemia reperfusion injury and adhesion. Neurology 1997; 49: 2326.
Bevilacqua MP. Endothelial-leukocyte adhesion molecules. Annu Rev Immunol 1993; 11: 767804.
Panés J, Perry M, Granger N. Leukocyte endothelial cell adhesion: avenues for therapeutic intervention. Br J Pharmacol 1999; 126: 537550.
Chopp M, Zhang L, Chen H, Li Y, Jiang N, Rusche R. Post ischemic administration of an anti-MAC-1 antibody reduces ischemic cell damage after transient middle cerebral artery occlusion in the rat. Stroke 1994; 25: 869876.
Matsuo Y, Onodera H, Shiga Yet al. Correlation between myeloperoxidase-quantified neutrophil accumulation and ischemic brain injury in rat: effects of neutrophil depletion. Stroke 1994; 25: 14691475.
Zhang L, Chopp M, Jiang N, Tang X, Prostak J, Manning M. Anti-ICAM-1 antibody reduces ischemic cell damage after transient but not permanent MCA occlusion in the Wistar rat. Stroke 1995; 926: 14381443.
Haefely E. Central actions of benzodiazepines: general introduction. Br J Psychiatry 1978; 133: 231238.
Bond PA, Cundall RL, Rolfe B. [3H] Diazepam binding to human granulocytes. Life Sciences 1985; 37: 1116.


Effect of midazolam on in vitro cerebral endothelial ICAM-1 expression induced by astrocyte-conditioned medium

  • K. Ghori (a1), D. Harmon (a1), F. Walsh (a1) and G. Shorten (a1)


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