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Overexpressed α7 nicotinic acetylcholine receptor inhibited proinflammatory cytokine release in NIH3T3 cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
22052 43251 2009 7 PDF Available
Title
Overexpressed α7 nicotinic acetylcholine receptor inhibited proinflammatory cytokine release in NIH3T3 cells
Abstract

Alpha-7 nicotinic acetylcholine receptor (α7nAChR) played an important role during the process of inflammation, and was a potential therapeutic target for many diseases, including Alzheimer's disease, endotoxaemia, hypertension etc. However, it was difficult to express α7nAChR on commonly used host cells because of the lack of the chaperone protein RIC-3, which was indispensable to promote folding, assembly, and surface expression of α7nAChR. This work was designed to develop a cell line, which not only expressed α7nAChR highly and stably, but possessed biological activity. Full-length α7nAChR gene was extracted from rat peritoneal macrophages, recombinant plasmid pIRES2-EGFP-α7nAChR was constructed, and then transfected to the mouse fibroblast cell line NIH3T3, a cell line expressing RIC-3. Compared with the NIH3T3-pIRES2-EGFP, expression of α7nAChR was significantly higher in NIH3T3-pIRES2-EGFP-α7nAChR on both mRNA and protein levels. To evaluate the anti-inflammation activity of the over-expressed α7nAChR, TNF-α, IL-6, and IL-1β release induced by lipopolysaccharide was determined. No changes were found between NIH3T3-pIRES2-EGFP and NIH3T3-pIRES2-EGFP-α7nAChR culture supernatant under LPS challenge, however under nicotine pre-stimulation, pro-inflammatory cytokine release was significantly attenuated in α7nAChR transfected cell line. Moreover, in this established cell line, PNU-282987, a selective α7nAChR agonist, exerted a stronger ability to reduce TNF-α release than nicotine. We concluded that a functional NIH3T3-pIRES2-EGFP-α7nAChR cell line was developed, which might be useful for agonist screen and biological research.

Keywords
α7 nicotinic acetylcholine receptor; NIH3T3 cells; Lipopolysaccharide; Antiinflammation; Transfection
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Overexpressed α7 nicotinic acetylcholine receptor inhibited proinflammatory cytokine release in NIH3T3 cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 108, Issue 2, August 2009, Pages 85–91
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us