Skip to main content Accessibility help

Midazolam attenuates adenosine diphosphate-induced P-selectin expression and platelet–leucocyte aggregation

  • C. S. Tsai (a1), P. C. Hsu (a1), G. S. Huang (a2), T. C. Lin (a2), G. J. Hong (a1), C. M. Shih (a3) and C. Y. Li (a2)...



Background and objective: The expression of P-selectin on the surface of platelets and platelet–leucocyte conjugate formation are considered to be an indicator of platelet activation in thrombotic and inflammatory disease. Midazolam is a widely used sedative and anaesthetic induction agent. It may inhibit platelet aggregation and suppress interleukin-6 and -8 response in human leucocytes, but any effect on the adhesion of activated platelets to leucocytes remains obscure. We have examined the influence of midazolam on adenosine diphosphate (ADP)-induced platelet surface P-selectin expression and platelet–leucocyte aggregation in whole blood.

Methods: Human whole blood was stimulated with 2 × 10−5M ADP in the presence of midazolam (3 × 10−4 to 3 × 10−6M). Samples were stained with a fluorochrome-conjugated CD62P and CD41a antibody for detecting human platelet P-selectin antigens. The leucocyte subpopulations were separately gated and platelet–leucocyte aggregates were defined as cells found positive for CD45 and CD62P. All samples were analysed and were electronically separated into specific cell types (platelets, neutrophils, monocytes and lymphocytes) according to their typical forward/side scattering by flow cytometry.

Results: Midazolam significantly inhibited ADP-induced platelet P-selectin expression and attenuated platelet–leucocyte aggregation (mainly in neutrophils and monocytes) in a dose-dependent manner with a maximum inhibitory effect at 3 × 10−4M (P < 0.01).

Conclusions: This study demonstrated that midazolam decreases the ADP-induced expression of platelet surface P-selectin and platelet–leucocyte aggregation.


Corresponding author

Correspondence to: Chi-Yuan Li, Department of Anesthesiology, Tri-Service General Hospital, #325, Section 2, Cheng-Gung Road, Taipei, Taiwan, ROC. E-mail:; Tel: +886 2 8792 7128; Fax: +886 2 8792 7127


Hide All


Harrison P, Cramer EM. Platelet alpha-granules. Blood Rev 1993; 7: 5262.
Rinder HM, Bonan JL, Rinder CS, Ault KA, Smith BR. Dynamics of leukocyte–platelet adhesion in whole blood. Blood 1991; 78: 17301737.
Hollander JE, Muttreja MR, Dalesandro MR, Shofer FS. Risk stratification of emergency department patients with acute coronary syndromes using P-selectin. J Am Coll Cardiol 1999; 34: 95105.
Kamath S, Blann AD, Caine GJ, Gurney D, Chin BS, Lip GY. Platelet P-selectin levels in relation to plasma soluble P-selectin and beta-thromboglobulin levels in atrial fibrillation. Stroke 2002; 33: 12371242.
Hofbauer R, Moser D, Hammerschmidt V, Kapiotis S, Frass M. Ketamine significantly reduces the migration of leukocytes through endothelial cell monolayers. Crit Care Med 1998; 26: 15451549.
Dundee JW, Halliday NJ, Harper KW, Brogden RN. Midazolam. A review of its pharmacological properties and therapeutic use. Drugs 1984; 28: 519543.
Sheu JR, Hsiao G, et al. Mechanisms involved in the antiplatelet activity of midazolam in human platelets. Anesthesiology 2002; 96: 651658.
Miyawaki T, Sogawa N, Maeda S, Kohjitani A, Shimada M. Effect of midazolam on interleukin-6 mRNA expression in human peripheral blood mononuclear cells in the absence of lipopolysaccharide. Cytokine 2001; 15: 320327.
Galley HF, Dubbels AM, Webster NR. The effect of midazolam and propofol on interleukin-8 from human polymorphonuclear leukocytes. Anesth Analg 1998; 86: 12891293.
Ramoska EA, Linkenheimer R, Glasgow C. Midazolam use in the emergency department. J Emerg Med 1991; 9: 247251.
Michelson AD, Barnard MR, et al. In vivo tracking of platelets: circulating degranulated platelets rapidly lose surface P-selectin but continue to circulate and function. Proc Natl Acad Sci USA 1996; 93: 1187711882.
Field EJ. Platelet adherence to polymorphs. BMJ 1963; 2: 388.
Mickelson JK, Lakkis NM, Villarreal-Levy G, Hughes BJ, Smith CW. Leukocyte activation with platelet adhesion after coronary angioplasty: a mechanism for recurrent disease? J Am Coll Cardiol 1996; 28: 345353.
Hill JD, Ratliff JL, et al. Pulmonary pathology in acute respiratory insufficiency: lung biopsy as a diagnostic tool. J Thorac Cardiovasc Surgery 1976; 71: 6471.
Weiss HJ. Inherited abnormalities of platelet granules and signal transduction. In: Colman RW, Hirsh J, Marder VJ, Salzman EW, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice, 3rd edn. Philadelphia, USA: Lippincott, 1994: 673684.
Cattaneo M, Lombardi R, et al. Deficiency of (33P)2MeS-ADP-binding sites on platelets with secretion defect, normal granule stores and normal thromboxane A2 production. Evidence that ADP potentiates platelet secretion independently of the formation of large platelet aggregates and thromboxane A2 production. Thromb Haemost 1997; 77: 986990.
Cattaneo M, Lecchi A, Lombardi R, Gachet C, Zighetti ML. Platelets from a patient heterozygous for the defect of P2CYC receptors for ADP have a secretion defect despite normal thromboxane A2 production and normal granule stores: further evidence that some cases of platelet ‘primary secretion defect’ are heterozygous for a defect of P2CYC receptors. Arterioscler Thromb Vasc Biol 2000; 20: E101E106.
Storey RF, Judge HM, Wilcox RG, Heptinstall S. Inhibition of ADP-induced P-selectin expression and platelet–leukocyte conjugate formation by clopidogrel and the P2Y12 receptor antagonist AR-C69931MX but not aspirin. Thromb Haemost 2002; 88: 488494.
Cattaneo M, Canciani MT, et al. Released adenosine diphosphate stabilizes thrombin-induced human platelet aggregates. Blood 1990; 75: 10811086.
Caron A, Theoret JF, Mousa SA, Merhi Y. Anti-platelet effects of GPIIb/IIIa and P-selectin antagonism, platelet activation, and binding to neutrophils. J Cardiovasc Pharmacol 2002; 40: 296306.
Jy W, Horstman LL, Park H, Mao WW, Valant P, Ahn YS. Platelet aggregates as markers of platelet activation: characterization of flow cytometric method suitable for clinical applications. Am J Hematol 1998; 57: 3342.
Li N, Hu H, Lindqvist M, Wikstrom-Jonsson E, Goodall AH, Hjemdahl P. Platelet–leukocyte cross talk in whole blood. Arterioscler Thromb Vasc Biol 2000; 20: 27022708.
Rinder HM, Tracey JL, Rinder CS, Leitenberg D, Smith BR. Neutrophil but not monocyte activation inhibits P-selectin-mediated platelet adhesion. Thromb Haemost 1994; 72: 750756.
Kirchhofer D, Riederer MA, Baumgartner HR. Specific accumulation of circulating monocytes and polymorphonuclear leukocytes on platelet thrombi in a vascular injury model. Blood 1997; 89: 12701278.
Celi A, Pellegrini G, et al. P-selectin induces the expression of tissue factor on monocytes. Proc Natl Acad Sci USA 1994; 91: 87678771.
Bonomini M, Stuard S, et al. Neutrophil reactive oxygen species production during hemodialysis: role of activated platelet adhesion to neutrophils through P-selectin. Nephron 1997; 75: 402411.
Furman MI, Benoit SE, et al. Increased platelet reactivity and circulating monocyte–platelet aggregates in patients with stable coronary artery disease. J Am Coll Cardiol 1998; 31: 352358.
Gawaz M, Bogner C. Changes in platelet membrane glycoproteins and platelet–leukocyte interaction during hemodialysis. Clin Invest 1994; 72: 424429.
Weyrich AS, McIntyre TM, McEver RP, Prescott SM, Zimmerman GA. Monocyte tethering by P-selectin regulates monocyte chemotactic protein-1 and tumor necrosis factor-alpha secretion. Signal integration and NF-kappa B translocation. J Clin Invest 1995; 95: 22972303.
Weyrich AS, Elstad MR, et al. Activated platelets signal chemokine synthesis by human monocytes. J Clin Invest 1996; 97: 15251534.
Colleoni M, Costa B, Gori E, Santagostino A. Biochemical characterization of the effects of the benzodiazepine, midazolam, on mitochondrial electron transfer. Pharmacol Toxicol 1996; 78: 6976.
Matsumoto T, Ogata M, Koga K, Shigematsu A. Effect of peripheral benzodiazepine receptor ligands on lipopolysaccharide-induced tumor necrosis factor activity in thioglycolate-treated mice. Antimicrob Agent Chemother 1994; 38: 812816.
Harper KW, Collier PS, Dundee JW, Elliott P, Halliday NJ, Lowry KG. Age and nature of operation influence the pharmacokinetics of midazolam. Br J Anaesth 1985; 57: 866871.
Larsen E, Celi A, et al. PADGEM protein: a receptor that mediates the interaction of activated platelets with neutrophils and monocytes. Cell 1989; 59: 305312.
Spangenberg P, Redlich H, Bergmann I, Losche W, Gotzrath M, Kehrel B. The platelet glycoprotein IIb/IIIa complex is involved in the adhesion of activated platelets to leukocytes. Thromb Haemost 1993; 70: 514521.


Midazolam attenuates adenosine diphosphate-induced P-selectin expression and platelet–leucocyte aggregation

  • C. S. Tsai (a1), P. C. Hsu (a1), G. S. Huang (a2), T. C. Lin (a2), G. J. Hong (a1), C. M. Shih (a3) and C. Y. Li (a2)...


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed