CAJ | 학술논문

Objective: To investigate the dose and timedependent effects of lipopolysaccharide ( LPS ) on cytoskeletal F-acitn and G-actin reorganizations by visualizing their distribution and measuring their contents in human umbilical vein endothelial cell line ECV-304. Methods: F-actin was labeled with rhodaminephalloidin and G-actin with deoxyribonuclease Ⅰ (DNase Ⅰ) conjugated with fiuorescein isothiocyanate ( FITC ). Contents of cytoskeletal proteins were obtained by flow cytometry.Results: F-actin was mainly distributed peripherally in endothelial cells under normal conditions. LPS stimulation caused the formation of stress fibers and filopodia. G-actin was normally seen in perinuclear and nuclear areas in control ECV-304 cells. Under LPS stimulation, G-actin dots appeared in the cytoplasmic region. The actin disorganization was accompanied by the time- and dose- dependent decrease in F-actin pool andincrease in G-actin pool.Conclusions: LPS can induce characteristic morphological alterations of actin cytoskeleton and formation of intercellular gap in endothelial cells,accompanied by changes in F-actin and G-actin pools.