Dextromethorphan Attenuates LPS-Induced Adhesion Molecule Expression in Human Endothelial Cells

Objective This study examines the effect of Dextromethorphan (d‐3‐methoxy‐17‐methylmorphinan; DXM), a commonly used cough‐suppressing drug, on the expression of VCAM‐1 and ICAM‐1 in human umbilical vein endothelial cells (HUVECs) stimulated with lipopolysaccharide (LPS). Methods The effect of DXM on...

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Published inMicrocirculation (New York, N.Y. 1994) Vol. 20; no. 2; pp. 190 - 201
Main Authors Jiang, Shinn-Jong, Hsu, Sheng-Yao, Deng, Chuan-Rou, Huang, Huey-Chun, Liu, Shu-Lin, Shi, Guey-Yueh, Wu, Hua-Lin
Format Journal Article
LanguageEnglish
Published United States Blackwell Publishing Ltd 01.02.2013
Wiley Subscription Services, Inc
Subjects
adhesion molecule
Anti-Inflammatory Agents - pharmacology
Cell adhesion & migration
Cell Communication - drug effects
Cell Movement - drug effects
Cells
dextromethorphan
Dextromethorphan - pharmacology
Drug Interactions
endothelial cell
Endothelial Cells - cytology
Endothelial Cells - drug effects
Endothelial Cells - metabolism
Excitatory Amino Acid Antagonists - pharmacology
Human Umbilical Vein Endothelial Cells
Humans
inflammation
Intercellular Adhesion Molecule-1 - metabolism
Kinases
Lipopolysaccharides - pharmacology
MAP Kinase Signaling System - drug effects
Monocytes - cytology
Monocytes - drug effects
Monocytes - metabolism
NF-kappa B - metabolism
Proteins
Proto-Oncogene Proteins c-akt - metabolism
Vascular Cell Adhesion Molecule-1 - metabolism
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Summary:Objective This study examines the effect of Dextromethorphan (d‐3‐methoxy‐17‐methylmorphinan; DXM), a commonly used cough‐suppressing drug, on the expression of VCAM‐1 and ICAM‐1 in human umbilical vein endothelial cells (HUVECs) stimulated with lipopolysaccharide (LPS). Methods The effect of DXM on expression of cell adhesion molecules induced by LPS was evaluated by monocyte bindings in vitro and ex vivo and transmigration assays. The signaling pathways involved in the inflammation inhibitory effect of DXM were analyzed by Western blot and immunofluorescent stain. Results Pretreatment of HUVECs with DXM inhibited LPS‐induced adhesion of THP‐1 cells in vitro and ex vivo, and reduced transendothelial migration of these cells. Furthermore, treatment of HUVECs with DXM can significantly decrease LPS‐induced expression of ICAM‐1 and VCAM‐1. DXM abrogated LPS‐induced phosphorylation of ERK and Akt. The translocation of early growth response gene‐1 (Egr‐1), a downstream transcription factor involved in the mitogen‐activated kinase (MEK)‐ERK signaling pathway, was suppressed by DXM treatment. Furthermore, DXM inhibited LPS‐induced IκBα degradation and nuclear translocation of p65. Conclusions Dextromethorphan inhibits the adhesive capacity of HUVECs by reducing the LPS‐induced ICAM‐1 and VCAM‐1 expression via the suppression of the ERK, Akt, and NF‐κB signaling pathways. Thus, DXM is a potential anti‐inflammatory therapeutic that may modulate atherogenesis.
Bibliography:National Science Council - No. NSC-99-2320-B-320-001
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istex:D1DAD6E8CA5E52662C4A2D32C5048476ACC4BFDA
Figure S1. DXM inhibits LPS-induced ICAM-1 and VCAM-1expression. HUVECs were pretreated with various concentrations of DXM and then combined with 100 ng/ml LPS for the indicated times. Total cell lysate was prepared, proteins were separated by SDS-PAGE and immunoblotting was performed using corresponding specific antibodies. Data are representative of three independent experiments with identical conditions.Figure S2. DXM attenuates LPS-induced signaling activation of Akt, ERK, and IkB-a protein in a dose-dependent manner. HUVECs were pretreated with various concentrations of DXM and then combined with 100 ng/ml LPS for the indicated time. Total cell lysate was prepared, proteins were separated by SDS-PAGE and immunoblotting was performed using corresponding specific antibodies. Data are representative of three independent experiments with identical conditions.Figure S3. DXM decreases LPS-induced transactivation of Egr-1 in HUVECs. HUVECs were pretreated with various concentrations of DXM for 1 h and then combined with 100 ng/ml LPS for 2 h. Nuclear extracts was harvested, the proteins were separated by SDS-PAGE, and the expression of Egr-1 was detected by western blot.Figure S4. DXM decreases LPS-induced transactivation of p65 in HUVECs. HUVECs were pretreated with various concentrations of DXM for 1 h and then combined with 100 ng/ml LPS for 2 h. Nuclear extracts was harvested, the proteins were separated by SDS-PAGE, and the expression of p65 was detected by western blot.Figure S5. Dextromethorphan decreases LPS-induced reactive oxygen species (ROS) production. Endothelial cells were pretreated with DXM or N-acetyleysteine (NAC) for one hour and then stimulated with LPS for 20 minutes. 5 μM Carboxy H2DCFDA was added and incubated for five minutes. ROS were detected by fluorescent microscopy and cells were quantified. The data are presented in mean ± SD (n = 3). ***p < 0.001 as compared to control; ###p < 0.001 as compared to cells stimulated with LPS in the absence of DXM.
ArticleID:MICC12024
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1073-9688
1549-8719
DOI:10.1111/micc.12024