Open Access 01-12-2013 | Research
Different actions of endothelin-1 on chemokine production in rat cultured astrocytes: reduction of CX3CL1/fractalkine and an increase in CCL2/MCP-1 and CXCL1/CINC-1
Published in: Journal of Neuroinflammation | Issue 1/2013
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Background
Chemokines are involved in many pathological responses of the brain.
Astrocytes produce various chemokines in brain disorders, but little is
known about the factors that regulate astrocytic chemokine production.
Endothelins (ETs) have been shown to regulate astrocytic functions through
ETB receptors. In this study, the effects of ETs on chemokine
production were examined in rat cerebral cultured astrocytes.
Methods
Astrocytes were prepared from the cerebra of one- to two-day-old Wistar rats
and cultured in serum-containing medium. After serum-starvation for 48
hours, astrocytes were treated with ETs. Total RNA was extracted using an
acid-phenol method and expression of chemokine mRNAs was determined by
quantitative RT-PCR. The release of chemokines was measured by ELISA.
Results
Treatment of cultured astrocytes with ET-1 and Ala1,3,11,15-ET-1,
an ETB agonist, increased mRNA levels of CCL2/MCP1 and
CXCL1/CINC-1. In contrast, CX3CL1/fractalkine mRNA expression decreased in
the presence of ET-1 and Ala1,3,11,15-ET-1. The effect of ET-1 on
chemokine mRNA expression was inhibited by BQ788, an ETB
antagonist. ET-1 increased CCL2 and CXCL1 release from cultured astrocytes,
but decreased that of CX3CL1. The increase in CCL2 and CXCL1 expression by
ET-1 was inhibited by actinomycin D, pyrrolidine dithiocarbamate, SN50,
mithramycin, SB203580 and SP600125. The decrease in CX3CL1 expression by
ET-1 was inhibited by cycloheximide, Ca2+ chelation and
staurosporine.
Conclusion
These findings suggest that ETs are one of the factors regulating astrocytic
chemokine production. Astrocyte-derived chemokines are involved in
pathophysiological responses of neurons and microglia. Therefore, the
ET-induced alterations of astrocytic chemokine production are of
pathophysiological significance in damaged brains.