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Morphine down regulates human vascular tissue estrogen receptor expression determined by real-time RT-PCR
Patrick Cadet1, Kirk Mantione, Thomas V Bilfinger
1Neuroscience Research Institute, State University of New York at Old Westbury, NY 11568, USA.
Unlabelled:
Human vascular endothelial cells express the estrogen receptor-beta (ER-beta), which can be modulated by the opiate alkaloid morphine.
Objectives:
To determine if morphine is capable of down regulating the ER-beta receptor in a similar fashion as the mu opiate receptor since they are both coupled to constitutive nitric oxide synthase derived nitric oxide release.
Methods And Results:
Endothelial cells obtain from human vascular tissues (saphenous vein, atria and primary saphenous vein cells) were treated with 1 uM morphine plus or minus the mu opiate receptor antagonist naloxone or CTOP (10 uM) for 24 h at 37 degrees C. Total RNA was isolated from treated and untreated primary endothelial cells, and specific primers and a probe were used to determine the ER-beta gene expression by real-time RT-PCR. Cells treated with morphine exhibited a down-regulation of ER-beta, whereas naloxone and CTOP were able to partially block the morphine effect. In addition, the 266 bp fragment generated by RT-PCR using the same primers as in the real-time PCR was sequenced and revealed a 100% sequence identity as the authentic ER-beta gene sequence.
Conclusions:
These results indicate that ER-beta is expressed in human vascular endothelial cells, and morphine appears to regulate this receptor in a similar fashion as the mu opiate receptor.