Abstract
Vascular cell adhesion molecule-1 (VCAM-1) is expressed in early stages of atherosclerosis: however, the mechanisms of its upregulation are not fully understood. In the present study, we examined the effects of interleukin-4 (IL-4) on VCAM-1 gene expression and its transcriptional regulatory mechanism in human umbilical vein endothelial cells (HUVEC). Reverse transcription-polymerase chain reaction showed that VCAM-1 mRNA was induced in IL-4-treated HUVEC in a time- and close-dependent manner. Among known transcription factors that have binding sites in the promoter region of the VCAM-1 gene, IL-4 activated only SP-1. In contrast, nuclear factor-κB (NF-κB), activator protein-1 (AP-1) and interferon regulatory factor-1 (IRF-1), which also have consensus binding sequences in the 5′-flanking region of the human VCAM-1 gene, were not activated. The role of SP-1 in IL-4-induced VCAM-1 expression was confirmed in HUVEC transfected with a reporter construct of the VCAM-1 promoter with mutated SP-1 binding site. As IL-4 treatment of HUVEC enhanced the intracellular oxidizing potential, as indicated by an increase in 2′,7′-dichlorofluorescein (DCF) fluorescence, we studied the effect of antioxidants on IL-4-induced VCAM-1 expression. Pretreatment of HUVEC with pyrrolidine dithiocarbamate (PDTC) or N-acetylcysteine (NAC) completely prevented IL-4-induced VCAM-1 expression. In addition, PDTC inhibited IL-4-related activation of SP-1. These results suggest that IL-4-induced oxidative stress upregulates the expression of VCAM-1 gene in HUVEC at transcriptional levels via activation of SP-1 transcription factor. In contrast, NF-κB, AP-1 or IRF-1 do not appear to be involved in the signal transduction cascade.
| Original language | English |
|---|---|
| Pages (from-to) | 83-94 |
| Number of pages | 12 |
| Journal | Journal of Molecular and Cellular Cardiology |
| Volume | 33 |
| Issue number | 1 |
| DOIs | |
| State | Published - Feb 19 2001 |
| Externally published | Yes |
Fingerprint
Keywords
- Atherosclerosis
- Human endothelial cells
- Interleukin-4
- Oxidative stress
- Transcriptional regulation
- Vascular cell adhesion molecule-1
ASJC Scopus subject areas
- Molecular Biology
- Cardiology and Cardiovascular Medicine
Cite this
IL-4-induced oxidative stress upregulates VCAM-1 gene expression in human endothelial cells. / Lee, Yong Woo; Kühn, Hartmut; Hennig, Bernhard; Neish, Andrew S.; Toborek, Michal J.
In: Journal of Molecular and Cellular Cardiology, Vol. 33, No. 1, 19.02.2001, p. 83-94.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - IL-4-induced oxidative stress upregulates VCAM-1 gene expression in human endothelial cells
AU - Lee, Yong Woo
AU - Kühn, Hartmut
AU - Hennig, Bernhard
AU - Neish, Andrew S.
AU - Toborek, Michal J
PY - 2001/2/19
Y1 - 2001/2/19
N2 - Vascular cell adhesion molecule-1 (VCAM-1) is expressed in early stages of atherosclerosis: however, the mechanisms of its upregulation are not fully understood. In the present study, we examined the effects of interleukin-4 (IL-4) on VCAM-1 gene expression and its transcriptional regulatory mechanism in human umbilical vein endothelial cells (HUVEC). Reverse transcription-polymerase chain reaction showed that VCAM-1 mRNA was induced in IL-4-treated HUVEC in a time- and close-dependent manner. Among known transcription factors that have binding sites in the promoter region of the VCAM-1 gene, IL-4 activated only SP-1. In contrast, nuclear factor-κB (NF-κB), activator protein-1 (AP-1) and interferon regulatory factor-1 (IRF-1), which also have consensus binding sequences in the 5′-flanking region of the human VCAM-1 gene, were not activated. The role of SP-1 in IL-4-induced VCAM-1 expression was confirmed in HUVEC transfected with a reporter construct of the VCAM-1 promoter with mutated SP-1 binding site. As IL-4 treatment of HUVEC enhanced the intracellular oxidizing potential, as indicated by an increase in 2′,7′-dichlorofluorescein (DCF) fluorescence, we studied the effect of antioxidants on IL-4-induced VCAM-1 expression. Pretreatment of HUVEC with pyrrolidine dithiocarbamate (PDTC) or N-acetylcysteine (NAC) completely prevented IL-4-induced VCAM-1 expression. In addition, PDTC inhibited IL-4-related activation of SP-1. These results suggest that IL-4-induced oxidative stress upregulates the expression of VCAM-1 gene in HUVEC at transcriptional levels via activation of SP-1 transcription factor. In contrast, NF-κB, AP-1 or IRF-1 do not appear to be involved in the signal transduction cascade.
AB - Vascular cell adhesion molecule-1 (VCAM-1) is expressed in early stages of atherosclerosis: however, the mechanisms of its upregulation are not fully understood. In the present study, we examined the effects of interleukin-4 (IL-4) on VCAM-1 gene expression and its transcriptional regulatory mechanism in human umbilical vein endothelial cells (HUVEC). Reverse transcription-polymerase chain reaction showed that VCAM-1 mRNA was induced in IL-4-treated HUVEC in a time- and close-dependent manner. Among known transcription factors that have binding sites in the promoter region of the VCAM-1 gene, IL-4 activated only SP-1. In contrast, nuclear factor-κB (NF-κB), activator protein-1 (AP-1) and interferon regulatory factor-1 (IRF-1), which also have consensus binding sequences in the 5′-flanking region of the human VCAM-1 gene, were not activated. The role of SP-1 in IL-4-induced VCAM-1 expression was confirmed in HUVEC transfected with a reporter construct of the VCAM-1 promoter with mutated SP-1 binding site. As IL-4 treatment of HUVEC enhanced the intracellular oxidizing potential, as indicated by an increase in 2′,7′-dichlorofluorescein (DCF) fluorescence, we studied the effect of antioxidants on IL-4-induced VCAM-1 expression. Pretreatment of HUVEC with pyrrolidine dithiocarbamate (PDTC) or N-acetylcysteine (NAC) completely prevented IL-4-induced VCAM-1 expression. In addition, PDTC inhibited IL-4-related activation of SP-1. These results suggest that IL-4-induced oxidative stress upregulates the expression of VCAM-1 gene in HUVEC at transcriptional levels via activation of SP-1 transcription factor. In contrast, NF-κB, AP-1 or IRF-1 do not appear to be involved in the signal transduction cascade.
KW - Atherosclerosis
KW - Human endothelial cells
KW - Interleukin-4
KW - Oxidative stress
KW - Transcriptional regulation
KW - Vascular cell adhesion molecule-1
UR - http://www.scopus.com/inward/record.url?scp=0035140619&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035140619&partnerID=8YFLogxK
U2 - 10.1006/jmcc.2000.1278
DO - 10.1006/jmcc.2000.1278
M3 - Article
VL - 33
SP - 83
EP - 94
JO - Journal of Molecular and Cellular Cardiology
JF - Journal of Molecular and Cellular Cardiology
SN - 0022-2828
IS - 1
ER -