摘要
Objective: To investigate the dose and time-dep endent 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 rhodamine-phalloidin and G -actin with deoxyribonuclease I (DNase I)conjugated with fluorescein isothiocya nate (FITC). Contents of cytoskeletal proteins were obtained by flow cytometry. Results: F-actin was mainly distributed peripherally in endoth elial cells under normal conditions. LPS stimulation caused the formation of str ess 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 i n the cytoplasmic region. The actin disorganization was accompanied by the time - and dose- dependent decrease in F-actin pool and increase in G-actin pool . Conclusions: LPS can induce characteristic morphological altera tions of actin cytoskeleton and formation of intercellular gap in endothelial ce lls,accompanied by changes in F-actin and G-actin pools.
基金
SupportedbytheNationalNaturalScienceFoundationofChina (No .30 0 2 80 0 8)andNationalFoundationofBasicScienceResearch (No .G2 0 0 0 0 5 70 0 4andG19990 5 4 2 0 2 )
