Abstract
The adhesion of human blood platelets is studied with an in vitro model using reflection contrast microscopy and an image analysis system. The adhesive feature is promoted by oxidatively modified low density lipoprotein, which also induces functional morphological changes of platelets. However, when washed platelets are pretreated with 0.05 mM cytochalasin D, oxidized low density lipoprotein (100 μg/ml) causes a slower increase of the adhesion area (11.6 μm2/min) compared to untreated platelets (15.7 μm2/min) or platelets treated by oxidized low density lipoprotein alone (20.5 μm2/min, P<0.01). These results are supported by light transmission analysis and by transmission electron microscopy. Our experiments suggest that cytochalasin D inhibits the change of platelets in shape induced by oxidized low density lipoprotein, hinders the adhesion, but does not prevent the adhesion entirely.
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Andrews HE, Aitken JW, Hassall DG, Skinner VO, Bruck-Dorfer KR (1987) Intracellular mechanisms in the activation of human platelets by low-density lipoproteins. Biochem J 242:559–564
Ardlie NG, Selley ML, Simons LA (1989) Platelet activation by oxidatively modified low density lipoproteins. Atherosclerosis 76:117–124
Aviram M (1989) Modified forms of low density lipoprotein affect platelet aggregation in vitro. Thromb Res 53:561–567
Aviram M (1991) Effect of lipoproteins and platelets on macrophage cholesterol metabolism. In: Harris JR (ed) Blood Cell Biochemistry, Vol 2: Megakaryocytes, Platelets, Macrophages and Eosinophils. Plenum Press, New York, pp 179–208
Aviram M (1992) Platelets and the arterial wall lesion. Curr Opin Lipid 3:344–348
Aviram M (1993) Modified forms of low density lipoprotein and atherosclerosis. Atherosclerosis 98:1–9
Aviram M, Brook G (1987) Platelet acitvation by plasma lipoproteins. Prog Cardiovasc Dis 30:61–72
Aviram M, Fuhrman B, Maor I, Brook G (1992) Serotonin increases macrophage uptake of oxidized low density lipoprotein. Eur J Clin Chem Clin Biochem 30:55–61
Casella JF, Flanagan MD, Lin SC (1981) Cytochalasin D inhibits actin polymerization and induces depolymerization of actin filaments formed during platelet shape change. Nature 293:302–305
Filler TJ, Rickert CH, Fassnacht UK, Pera F (1994) Reflection contrast microscopy within chrome-alum haematoxylin stained tissue-sections. Histochemistry 101:375–378
Fuhrman B, Brook GJ, Aviram M (1992) Proteins derived from platelet α-granules modulate the uptake of oxidized low density lipoprotein by macrophages. Biochem Biophys Acta 1127:15–21
Horio T, Kohno M, Yasunari K, Murakawa K, Yokokawa K, Ikeda M, Fukui T, Takeda T (1993) Stimulation of endothelin-1 release by low density and very low density lipoproteins in cultured human endothelial cells. Atherosclerosis 101:185–190
Kishi Y, Numano F (1989) In vitro study of vascular endothelial injury by activated platelets and its prevention. Atherosclerosis 76:95–101
Koller E, Volf I, Koller F (1992) Binding of low density lipoprotein to human platelets: effects of chymotrypsin. Thromb Haemorrh Disorders 6/2:57–65
Kometani M, Sato T, Fujii T (1986) Platelet cytoskeletal components involved in shape change and secretion. Throm Res 41:801–809
Kugiyama K, Sakamoto T, Misumi I, Sugiyama S, Ohgushi M, Ogawa H, Horiguchi M, Yasue H (1993) Transferable lipids in oxidized low-density lipoprotein stimulate plasminogen activator inhibitor-1 and inhibit tissue-type plasminogen activator release from endothelial cells. Circ Res 73:335–343
Mackness MI, Abbott C, Arrol S, Durrington PN (1993) The role of high-density lipoprotein and lipid-soluble antioxidant vitamins in inhibiting low-density lipoprotein oxidation. Biochem J 294:829–834
Malle E, Sattler W (1994) Platelets and the lipoproteins: native, modified and platelet modified lipoproteins. Platelets 5:70–83
O'Halloran T, Beckerle MC, Burridge K (1985) Identification of talin as a major cytoplasmic protein implicated in platelet activation. Nature 317(6036):449–451
Olorundare OE, Simmons SR, Albrecht RM (1992) Cytochalasin D and E: effects on fibrinogen receptor movement and cytoskeletal reorganization in fully spread, surface-activated platelets: a correlative light and electron microscopic investigation. Blood 79:99–109
Ozaki Y, Jinnai Y, Yatomi Y, Kume S (1993) Protein kinase C inhibitors suppress disc-sphere changes of human platelets, as assessed with the shape-change parameter. Eur J Pharmacol 235:255–265
Palinski W, Rosenfeld ME, Herttuala SY, Gurtner GC, Socher SS, Butler SW, Parthasarathy S, Carew TE, Steinger D, Witztum JL (1989) Low density lipoprotein undergoes oxidative modification in vivo. Proc Natl Acad Sci USA 86:1372–1376
Patscheke H, Bubler D, Deranleau D, Lüscher EF (1984) Optical shape change analysis in stirred and unstirred human platelet suspensions. A comparison of aggregometric and stoppedflow turbidimetric measurements. Thromb Res 33:341–353
Pedreño J, Castellarnau C, Cullare C, Ortin R, Sanchez J, Llopart R and Gonzalez-Sastre F (1994) Platelet LDL receptor recognizes with the same apparent affinity both oxidized and native LDL-evidence that the receptor-ligand complexes are not internalized. Arterioscler Thromb 14:401–408
Petersen GL (1977) A simplification of the protein assay of Lowry et al. which is more generally applicable. Annal Biochem 83:346–363
Robenek H, Harrach B, Severs NJ (1991) Display of low density lipoprotein receptors is clustered, not dispersed, in fibroblast and hepatocyte plasma membranes. Arterioscler Thromb 11:261–271
Sherman CT, Litvack F, Grundfest W, Lee M, Hickey A, Chaux A, Kass R, Blanche C, Matloff J, Morgenstern L, Ganz W, Swan HJC, Forrester J (1986) Coronary angioscopy in patients with unstable angina pectoris. N Engl J Med 315:913–919
Siess W, Lapetina EG, Cuatrecasas P (1982) Cytochalasins inhibit arachidonic acid metabolism in thrombin-stimulated platelets. Biochemistry 79:7709–7713
Sinzinger H (1986) Role of platelets in atherosclerosis. Semin Thromb Haemostasis 12:124–133
Thomas JP, Geiger PG, Girotti AW (1993) Lethal damage to endothelial cells by oxidized low density lipoprotein: role of selenoperoxidases in cytoprotection against lipid hydroperoxide-and iron-mediated reactions. J Lipid Res 34:479–490
Willemse F, Nap M, Dekok LB, Eggink HF (1993) Image analysis in immunohistochemistry. Factors with a possible influence on the performance of VIDAS version 2.0, a commercially available true color image analysis system. Analyt Quant Cytol Histol 15:136–143
Zhao B, Dierichs R, Ahonen-Sann R (1993) Low concentration of LDL enhances platelet reactivity in vitro — a morphological study. Platelets 4:41–44
Zhao B, Dierichs R, Liu B, Berkes P (1994a) Gold-labeled low density lipoproteins bind to washed human platelets. Platelets 5:113–120
Zhao B, Dierichs R, Liu B, Holling-Rauß M (1994b) Functional morphological alterations of human blood platelets induced by oxidized low density lipoprotein. Thromb Res 74:293–301
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Zhao, B., Filler, T.J., Rickert, C.H. et al. Decreased adhesion of oxidized LDL-stimulated platelets caused by cytochalasin D. Cell Tissue Res 280, 183–188 (1995). https://doi.org/10.1007/BF00304523
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DOI: https://doi.org/10.1007/BF00304523