title: Upregulating nonneuronal cholinergic activity decreases TNF release from lipopolysaccharide-stimulated RAW264.7 cells.
abstract: Nonneuronal cholinergic system plays a primary role in maintaining homeostasis. It has been proved that endogenous neuronal acetylcholine (ACh) could play an anti-inflammatory role, and exogenous cholinergic agonists could weaken macrophages inflammatory response to lipopolysaccharide (LPS) stimulation through activation of α7 subunit-containing nicotinic acetylcholine receptor (α7nAChR). We assumed that nonneuronal cholinergic system existing in macrophages could modulate inflammation through autocrine ACh and expressed α7nAChR on the cells. Therefore, we explored whether LPS continuous stimulation could upregulate the nonneuronal cholinergic activity in macrophages and whether increasing autocrine ACh could decrease TNF release from the macrophages. The results showed that, in RAW264.7 cells incubated with LPS for 20 hours, the secretion of ACh was significantly decreased at 4 h and then gradually increased, accompanied with the enhancement of α7nAChR expression level. The release of TNF was greatly increased from RAW264.7 cells at 4 h and 8 h exposure to LPS; however, it was suppressed at 20 h. Upregulating choline acetyltransferase (ChAT) expression through ChAT gene transfection could enhance ACh secretion and reduce TNF release from the infected RAW264. 7cells. The results indicated that LPS stimulation could modulate the activity of nonneuronal cholinergic system of RAW264.7 cells. Enhancing autocrine ACh production could attenuate TNF release from RAW264.7 cells.
authors.affiliation: Laboratory of Shock and Multiple Organ Dysfunction, Burns Institute, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lv
authors.affiliation: Laboratory of Shock and Multiple Organ Dysfunction, Burns Institute, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Hu
authors.affiliation: Department of Pathology, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Jiangyang
authors.initials: J
authors.lastname: Lu
authors.affiliation: Laboratory of Shock and Multiple Organ Dysfunction, Burns Institute, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Dong
authors.affiliation: Department of Pathology, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Laboratory of Shock and Multiple Organ Dysfunction, Burns Institute, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Minghua
authors.initials: M
authors.lastname: Du
authors.affiliation: Laboratory of Shock and Multiple Organ Dysfunction, Burns Institute, First Hospital Affiliated to the People's Liberation Army General Hospital, 51 Fu Cheng Road, Beijing 100048, China.
authors.forename: Huiping
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002460:Cell Line; D002469:Cell Separation; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D005434:Flow Cytometry; D008070:Lipopolysaccharides / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000378:metabolism*
chemical_list: D000893:Anti-Inflammatory Agents; D003470:Culture Media; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D064569:alpha7 Nicotinic Acetylcholine Receptor; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine
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