Possible involvement of Sphingosine-1-phosphate/Gi/RhoA pathways in adherence of eosinophils to pulmonary endothelium

Toyokazu Sashio, Hiroaki Kume, Naoya Takeda, Toshiaki Asano, Seita Tsuji, Masashi Kondo, Yoshinori Hasegawa, Kaoru Shimokata

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background: Sphingosine-1-phosphate (S1P), a lysophospholipid released from inflammatory cells, causes cell migration by increasing cytokines and chemokines. This study was designed to determine whether S1P causes adherence of eosinophils to pulmonary endothelial cells via enhancement of adhesion molecule expression. Methods: Expression of VCAM-1 and ICAM-1 was assessed by RT-PCR and Western blot analysis in human pulmonary microvasucular endothelial cells (HPMVECs). The number of adherent eosinophils to HPMVECs was calculated according to adhesion assay. Results: Pre-treatment of HPMVECs with S1P increased mRNA and protein expression of VCAM-1, in contrast, did not dramatically increase those expression of ICAM-1. The maximal expression of these adhesion molecules in mRNA and protein was observed 4 and 8 h after exposure to S1P, respectively. Pre-treatment with S1P also activated RhoA, a monomeric G protein; the ability of S1P to enhance the expression of VCAM-1 was attenuated by RhoA related inhibitors such as Y-27632, C3 exoenzyme, and GGTI-286. The effects of S1P on VCAM-1 were attenuated by pre-incubation with pertussis toxin, which catalyzes the ADP-ribosylation of Gi, a heterotrimeric G protein. After HPMVECs were treated with S1P, adhesion of human eosinophilic leukemic cell line (EoL-1) cells to HPMVECs was enhanced in a concentration-dependent manner. Augmented adherence of EoL-1 cells by S1P was also attenuated by Y-27632 and pertussis toxin. S1P causes adherence of eosinophils to pulmonary endothelium via RhoA activation. Conclusions: S1P may act as a lipid mediator in asthma. The RhoA/Rho-kinase pathway may be a therapeutic target for preventing eosinophil infiltration to the airway.

Original languageEnglish
Pages (from-to)283-293
Number of pages11
JournalAllergology International
Volume61
Issue number2
DOIs
Publication statusPublished - 01-01-2012

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Eosinophils
Endothelium
Lung
Endothelial Cells
Vascular Cell Adhesion Molecule-1
Pertussis Toxin
Intercellular Adhesion Molecule-1
sphingosine 1-phosphate
Lysophospholipids
Heterotrimeric GTP-Binding Proteins
rho-Associated Kinases
Messenger RNA
Monomeric GTP-Binding Proteins
Chemokines
Adenosine Diphosphate
Cell Movement
Proteins
Asthma
Western Blotting
Cytokines

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy

Cite this

Sashio, Toyokazu ; Kume, Hiroaki ; Takeda, Naoya ; Asano, Toshiaki ; Tsuji, Seita ; Kondo, Masashi ; Hasegawa, Yoshinori ; Shimokata, Kaoru. / Possible involvement of Sphingosine-1-phosphate/Gi/RhoA pathways in adherence of eosinophils to pulmonary endothelium. In: Allergology International. 2012 ; Vol. 61, No. 2. pp. 283-293.
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Possible involvement of Sphingosine-1-phosphate/Gi/RhoA pathways in adherence of eosinophils to pulmonary endothelium. / Sashio, Toyokazu; Kume, Hiroaki; Takeda, Naoya; Asano, Toshiaki; Tsuji, Seita; Kondo, Masashi; Hasegawa, Yoshinori; Shimokata, Kaoru.

In: Allergology International, Vol. 61, No. 2, 01.01.2012, p. 283-293.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Possible involvement of Sphingosine-1-phosphate/Gi/RhoA pathways in adherence of eosinophils to pulmonary endothelium

AU - Sashio, Toyokazu

AU - Kume, Hiroaki

AU - Takeda, Naoya

AU - Asano, Toshiaki

AU - Tsuji, Seita

AU - Kondo, Masashi

AU - Hasegawa, Yoshinori

AU - Shimokata, Kaoru

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N2 - Background: Sphingosine-1-phosphate (S1P), a lysophospholipid released from inflammatory cells, causes cell migration by increasing cytokines and chemokines. This study was designed to determine whether S1P causes adherence of eosinophils to pulmonary endothelial cells via enhancement of adhesion molecule expression. Methods: Expression of VCAM-1 and ICAM-1 was assessed by RT-PCR and Western blot analysis in human pulmonary microvasucular endothelial cells (HPMVECs). The number of adherent eosinophils to HPMVECs was calculated according to adhesion assay. Results: Pre-treatment of HPMVECs with S1P increased mRNA and protein expression of VCAM-1, in contrast, did not dramatically increase those expression of ICAM-1. The maximal expression of these adhesion molecules in mRNA and protein was observed 4 and 8 h after exposure to S1P, respectively. Pre-treatment with S1P also activated RhoA, a monomeric G protein; the ability of S1P to enhance the expression of VCAM-1 was attenuated by RhoA related inhibitors such as Y-27632, C3 exoenzyme, and GGTI-286. The effects of S1P on VCAM-1 were attenuated by pre-incubation with pertussis toxin, which catalyzes the ADP-ribosylation of Gi, a heterotrimeric G protein. After HPMVECs were treated with S1P, adhesion of human eosinophilic leukemic cell line (EoL-1) cells to HPMVECs was enhanced in a concentration-dependent manner. Augmented adherence of EoL-1 cells by S1P was also attenuated by Y-27632 and pertussis toxin. S1P causes adherence of eosinophils to pulmonary endothelium via RhoA activation. Conclusions: S1P may act as a lipid mediator in asthma. The RhoA/Rho-kinase pathway may be a therapeutic target for preventing eosinophil infiltration to the airway.

AB - Background: Sphingosine-1-phosphate (S1P), a lysophospholipid released from inflammatory cells, causes cell migration by increasing cytokines and chemokines. This study was designed to determine whether S1P causes adherence of eosinophils to pulmonary endothelial cells via enhancement of adhesion molecule expression. Methods: Expression of VCAM-1 and ICAM-1 was assessed by RT-PCR and Western blot analysis in human pulmonary microvasucular endothelial cells (HPMVECs). The number of adherent eosinophils to HPMVECs was calculated according to adhesion assay. Results: Pre-treatment of HPMVECs with S1P increased mRNA and protein expression of VCAM-1, in contrast, did not dramatically increase those expression of ICAM-1. The maximal expression of these adhesion molecules in mRNA and protein was observed 4 and 8 h after exposure to S1P, respectively. Pre-treatment with S1P also activated RhoA, a monomeric G protein; the ability of S1P to enhance the expression of VCAM-1 was attenuated by RhoA related inhibitors such as Y-27632, C3 exoenzyme, and GGTI-286. The effects of S1P on VCAM-1 were attenuated by pre-incubation with pertussis toxin, which catalyzes the ADP-ribosylation of Gi, a heterotrimeric G protein. After HPMVECs were treated with S1P, adhesion of human eosinophilic leukemic cell line (EoL-1) cells to HPMVECs was enhanced in a concentration-dependent manner. Augmented adherence of EoL-1 cells by S1P was also attenuated by Y-27632 and pertussis toxin. S1P causes adherence of eosinophils to pulmonary endothelium via RhoA activation. Conclusions: S1P may act as a lipid mediator in asthma. The RhoA/Rho-kinase pathway may be a therapeutic target for preventing eosinophil infiltration to the airway.

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