Injury to cultured endothelial cells by thrombin-stimulated platelets

Journal Article · Tue Apr 01 04:00:00 EST 1986 · Lab. Invest.; (United States)

OSTI ID:5692470

Jorgensen, L; Grothe, A G; Larsen, T; Kinlough-Rathbone, R L; Mustard, J F

In vivo, stimulated platelets may injure the endothelium. We have used cultured endothelial cells to assess endothelial cell damage caused by platelet stimulation with thrombin. Endothelial cells were cultured from umbilical veins and semiconfluent cultures were labeled with Na/sub 2/ /sup 51/CrO/sub 4/. Twenty four hours later washed human platelets (final concentration 200,000 platelets/microliters) and thrombin (final concentration 4 units/ml) were added to the medium and the culture dish was shaken for 15 minutes. The percentage of cells detached from the culture dish and the percentage of /sup 51/Cr lost from the endothelial cells into the ambient fluid during the shaking were determined and used as indicators of cell injury. Increased percentages of loosened cells and /sup 51/Cr in the ambient fluid were observed with platelet suspension and thrombin compared to controls with neither platelet suspension nor thrombin and controls with either platelet suspension or thrombin. The platelet-free supernatant obtained after reaction of the platelets with thrombin also increased the percentage of loosened cells, but it did not increase the percentage of /sup 51/Cr in the ambient fluid to a significant degree. Thrombin alone caused a moderate loss of /sup 51/Cr, but no increased loosening of cells. Treatment of the platelets with acetylsalicylic acid prior to the experiment depressed the detachment effect of thrombin-stimulated platelets, but did not alter the effect on the release of /sup 51/Cr into the ambient fluid. Scanning and transmission electron microscopy of cultured endothelial cells exposed to thrombin-stimulated platelets confirmed the presence of loosening and injury to the endothelial cells.

• Research Organization: McMaster Univ., Hamilton, Ontario
• OSTI ID: 5692470
• Journal Information: Lab. Invest.; (United States), Journal Name: Lab. Invest.; (United States) Vol. 4; ISSN LAINA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
BETA DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BLOOD VESSELS
BODY
BODY FLUIDS
CARDIOVASCULAR SYSTEM
CELL CULTURES
CHROMIUM 51
CHROMIUM ISOTOPES
COAGULANTS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTRON MICROSCOPY
ENDOTHELIUM
ENZYMES
EVEN-ODD NUCLEI
HEMATOLOGIC AGENTS
HEMOSTATICS
HYDROLASES
INJURIES
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
MATERIALS
MICROSCOPY
NUCLEI
ORGANS
PEPTIDE HYDROLASES
RADIOISOTOPES
SCANNING ELECTRON MICROSCOPY
SERINE PROTEINASES
STIMULATION
THROMBIN
TISSUES
TRACER TECHNIQUES
TRANSMISSION ELECTRON MICROSCOPY