Effects of lipopolysaccharide on actin reorganization and actin pools in endothelial cells.
- Author:
Qiao-bing HUANG
1
;
Li SONG
;
Ke-sen ZHAO
;
Bo CHEN
;
Xu-liang HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Actins; drug effects; Analysis of Variance; Cells, Cultured; Deoxyribonuclease I; Dose-Response Relationship, Drug; Endothelial Cells; chemistry; Escherichia coli; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Humans; Lipopolysaccharides; pharmacology; Phalloidine; Rhodamines; Umbilical Veins; cytology
- From: Chinese Journal of Traumatology 2004;7(4):195-200
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo investigate the dose and time-dependent 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.
METHODSF-actin was labeled with rhodamine-phalloidin and G-actin with deoxyribonuclease I (DNase I)conjugated with fluorescein isothiocyanate (FITC). Contents of cytoskeletal proteins were obtained by flow cytometry.
RESULTSF-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 and increase in G-actin pool.
CONCLUSIONSLPS 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.
