The Detection Of Platelet Microparticles And Their Effect Mechanism In The Thrombosis Formation

PARTâ… The Detection of Platelet Activation and its Morphological Change by Laser Scanning Confocal MicroscopyObjective To establish the method of detecting platelet activation and its morphological change directly by using the laser scanning confocal microscopy.Methods We induced platelet to activate with different concentration collagen, detected the fluorescence intensity change of platelet membrance glycoprotein (GP)â…¡b/â…¢a complex combining specific fluorescence (FITC/PE) antibody and judged the release of platelet microparticles, observed the difference of platelet morphology between the normal people and diabetic retinopathy by laser scanning confocal microscopy.Results Platelets were activated by inducer. We found the fluorescence intensity of the platelets labeled specific antibody changed in regular way. At certain concentrations, the addition of collagen induced the change of GPâ…¡b/â…¢a complex expression levels. And glycoprotein GPâ…¡b/â…¢a complex expression levels show increased tendency with the growth of collagen concentration. There were presence of pseudopodia and microparticles surrounding the platelet body or be near the terminal ends in diabetic retinopathy. They were different in platelet morphology as compared with normal people.Conclusion We can detect platelet membrance glycoprotein GPâ…¡b/â…¢a complex expression levels change because of platelets activation induced by collagen and release of platelet microparticles by using the laser scanning confocal microscopy. Meanwhile it is inferred observation of platelets morphology change have some value in diagnosis of platelets activation levels in thrombosis. PARTâ…¡The effect of platelet microparticles on the tissue factor expression of human umbilical vein endothelial cellsObjective To study whether platelet microparticles can induce human umbilical vein endothelial cells (ECV-304) expressing tissue factor and their effect mechanism in the thrombosis formation.Methods Some amount of platelet microparticles from human platelet rich plasma were incubated with ECV-304 for several hours. Semi-quantitative reverse transcription PCR was used to measure the expression of tissue factor mRNA of ECV-304 with platelet microparticles and without them, and flow cytometry was used to detect the tissue factor protein.Results Some amount of platelet microparticles can induce ECV-304 expressing tissue factor. And the tissue factor level treated by some amount of platelet microparticles was different from that treated by some concentration of lipopolysaccharide under the same reactive condition.Conclusion Some amount of platelet microparticles could induce ECV-304 expressing tissue factor in mRNA and protein level.