Acetylcholine regulation of synoviocyte cytokine expression by the alpha7 nicotinic receptor.
OBJECTIVE
The central nervous system can regulate peripheral inflammation, but the efferent neuronal routes and the mediators remain poorly defined. One candidate is the cholinergic pathway, which releases acetylcholine (ACh). This neurotransmitter can bind to the alpha7 cholinergic receptor (alpha7R) expressed by nonneuronal cells and reduce inflammation. To test this possibility, we evaluated the expression of alpha7R and its potential role as a target in rheumatoid arthritis (RA).


METHODS
The expression of alpha7R in human synovium and fibroblast-like synoviocytes (FLS) was determined using immunohistochemical, Western blot, and quantitative polymerase chain reaction (PCR) analyses. The effects of ACh in vitro were determined in interleukin-1 (IL-1)-stimulated FLS using immunoassays for protein, quantitative PCR for messenger RNA (mRNA), luciferase reporter constructs for IL-6 and NF-kappaB promoter activity, and electrophoretic mobility shift assays. Expression of alpha7R was knocked down with small interfering RNA (siRNA) or was inhibited with the selective alpha7R antagonist methyllycaconitine (MLA).


RESULTS
Protein and mRNA for alpha7R were demonstrated in RA and osteoarthritis synovium and cultured synoviocytes. Expression in synovium was mainly in the intimal lining. ACh significantly reduced the production of IL-6, CXCL8, CCL2, CCL3, CCL5, and granulocyte colony-stimulating factor by IL-1-stimulated FLS. This effect was blocked by the alpha7R antagonist MLA or by using alpha7R siRNA to knock down receptor expression. The selective alpha7R agonist PNU-282,987 decreased the production of IL-6 by IL-1-stimulated FLS. ACh did not reduce IL-6 transcription, but it decreased IL-6 mRNA half-life and reduced IL-6 mRNA steady-state levels.


CONCLUSIONS
The alpha7 receptor is expressed in the synovium and by synoviocytes. Receptor ligation inhibits cytokine expression in FLS through a posttranscriptional mechanism. Therefore, alpha7R is a potential therapeutic target for inflammatory diseases.
Waldburger
Jean-Marc
JM
University of California, San Diego, La Jolla, CA, USA.
Boyle
David L
DL
Pavlov
Valentin A
VA
Tracey
Kevin J
KJ
Firestein
Gary S
GS
D000109:Acetylcholine / Q000502:physiology*; D000157:Aconitine / Q000031:analogs & derivatives / Q000494:pharmacology; D001172:Arthritis, Rheumatoid / Q000378:metabolism; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western; D001643:Bridged Bicyclo Compounds / Q000494:pharmacology; D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis*; D006801:Humans; D007150:Immunohistochemistry; D015850:Interleukin-6 / Q000032:analysis; D016328:NF-kappa B / Q000032:analysis; D010003:Osteoarthritis / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D011978:Receptors, Nicotinic / Q000032:analysis / Q000502:physiology*; D013583:Synovial Membrane / Q000166:cytology*; D064569:alpha7 Nicotinic Acetylcholine Receptor
D001549:Benzamides; D001643:Bridged Bicyclo Compounds; C579167:Chrna7 protein, human; D016207:Cytokines; D015850:Interleukin-6; D016328:NF-kappa B; C498513:PNU-282987; D011506:Proteins; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; C054634:methyllycaconitine; D000109:Acetylcholine; D000157:Aconitine