Lipopolysaccharide induces expression of macrophage inflammatory protein-1alpha in human umbilical vein endothelial cells / 中华病理学杂志
Chinese Journal of Pathology
;
(12): 449-452, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-242154
ABSTRACT
<p><b>OBJECTIVE</b>To understand whether endotoxin lipopolysaccharide (LPS) is able to induce the expression of macrophage inflammatory protein-1alpha (MIP-1alpha) mRNA and protein in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>The expression of MIP-1alpha mRNA was determined by dot blotting analysis and by in situ hybridization using a digoxigenin-labeled MIP-1alpha cDNA probe after exposure of the cultured HUVECs to LPS at different concentrations. The expression of MIP-1alpha mRNA was determined by RT-PCR as well. In addition, the expression of MIP-1alpha protein was tested by cell enzyme-linked immunosorbent assay (ELISA) using a goat anti-human monoclonal MIP-1alpha antibody.</p><p><b>RESULTS</b>Dot blotting showed that the absorbance values of the dots on the nitrocellulose membrane were 1.490 and 3.310 when exposed to LPS at the concentrations of 1 micro g/ml and 10 micro g/ml which were 1.97- and 4.38-fold over that of the control group (0.775), respectively. In situ hybridization revealed that exposure to LPS at a concentration of 1 micro g/ml led to a significant increase in the MIP-1alpha mRNA expression in HUVECs as compared to the control group (F = 142.83, P < 0.01), whereas the MIP-1alpha mRNA in HUVECs was somewhat decreased when exposed to LPS at a concentration of 10 micro g/ml. RT-PCR revealed that the expression of MIP-1alpha mRNA in HUVECs were 1.65-, 2.86- and 1.26-fold over that of the control group when exposed to LPS at the concentrations of 1 micro g/ml, 5 micro g/ml and 10 micro g/ml respectively. Cell ELISA showed that after exposure of the HUVECs to LPS at the concentrations mentioned above, the expression of MIP-1alpha protein was strongly increased, especially in the 5 micro g/ml LPS group. Analysis of variance showed that there was a significant difference between groups (F = 15.36, P < 0.05).</p><p><b>CONCLUSIONS</b>LPS may induce a high level of MIP-1alpha mRNA and protein expression in HUVECs, and it might, hereby, play an important role in the recruitment of the monocytes/macrophages into the arterial intima.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Umbilical Veins
/
Enzyme-Linked Immunosorbent Assay
/
Cells, Cultured
/
Gene Expression Regulation
/
Lipopolysaccharides
/
In Situ Hybridization
/
Macrophage Inflammatory Proteins
/
Reverse Transcriptase Polymerase Chain Reaction
/
Endothelial Cells
/
Toxicity
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2003
Type:
Article
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