title: [Effect of nicotinic acetylcholine receptor α7 subunit gene on liver inflammatory reaction in mice with nonalcoholic steatohepatitis and related mechanisms].
abstract: Objective:To investigate the effect of nicotinic acetylcholine receptor α7 (α7nAChR) subunit gene on liver inflammation in mice with nonalcoholic steatohepatitis (NASH) and related mechanisms.Methods:C57BL/6J mice and α7nAChR gene knockout mice were fed for 24 weeks to establish the NASH model, and the mice were sacrificed to isolate and culture the primary liver macrophages. After the treatment with nicotine and endotoxin, ELISA was used to measure the levels of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in supernatant; indirect immunofluorescence assay and Western blot were used to observe the effect on the NF-κB signaling pathway, and quantitative PCR was used to measure the mRNA expression of Toll-like receptor-4 (TLR-4) in macrophages. An analysis of variance was used for comparison of means between multiple groups.Results:The results of ELISA showed that compared with the endotoxin+nicotine group of C57 NASH mice, the endotoxin+nicotine group of gene knockout NASH mice had significantly higher levels of IL-6 and TNF-α in supernatant (IL-6: 1 599±65 pg/ml vs 1 465±45 pg/ml,P< 0.05; TNF-α: 1 567±66 pg/ml vs 1 433±50 pg/ml,P< 0.05). The results of Western blot showed that compared with the endotoxin+nicotine group of C57 NASH mice, the endotoxin+nicotine group of gene knockout NASH mice had significantly higher relative protein expression of phosphorylated NF-κB and TLR-4 (NF-κB: 69 425±600 vs 51 133±200,P< 0.05; TLR-4: 93 387±684 vs 64 198±630,P< 0.05). The results of indirect immunofluorescence assay showed that the endotoxin+nicotine group of gene knockout NASH mice had a significantly higher fluorescence intensity of NF-κB than the endotoxin+nicotine group of C57 NASH mice. The results of PCR showed that the endotoxin+nicotine group of gene knockout NASH mice had significantly higher relative mRNA expression of TLR-4 than the endotoxin+nicotine group of C57 NASH mice (4.13±0.13 vs 2.93±0.14,P< 0.05).Conclusion:The α7nAChR gene knockout can aggravate the degree of inflammatory reaction in NASH, and its mechanism may be related to the fact that the NF-κB signaling pathway cannot be inhibited, which aggravates inflammatory reaction.
authors.affiliation: Department of Infectious Diseases, The Center Hospital of Shaoxing, Shaoxing 312030, China.
authors.forename: F Q
authors.initials: FQ
authors.lastname: Li
authors.affiliation: The Second Clinical Medial College, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors.forename: X C
authors.initials: XC
authors.lastname: Wu
authors.affiliation: The Second Clinical Medial College, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors.forename: L N
authors.initials: LN
authors.lastname: Xu
authors.affiliation: Department of Infectious Diseases, The Second Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310005, China.
authors.forename: X M
authors.initials: XM
authors.lastname: Chen
authors.affiliation: The Second Clinical Medial College, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors.forename: S
authors.initials: S
authors.lastname: Lu
authors.affiliation: Department of Infectious Diseases, The Second Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310005, China.
authors.forename: C L
authors.initials: CL
authors.lastname: Tang
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000627:therapeutic use*; D004195:Disease Models, Animal; D004731:Endotoxins / Q000627:therapeutic use*; D006505:Hepatitis; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D008099:Liver / Q000187:drug effects* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B; D009538:Nicotine / Q000627:therapeutic use*; D018722:Nicotinic Agonists / Q000627:therapeutic use*; D065626:Non-alcoholic Fatty Liver Disease / Q000188:drug therapy* / Q000209:etiology / Q000276:immunology; D010766:Phosphorylation; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000819:agonists* / Q000276:immunology
chemical_list: D000893:Anti-Inflammatory Agents; D004731:Endotoxins; D015850:Interleukin-6; D016328:NF-kappa B; D018722:Nicotinic Agonists; D014409:Tumor Necrosis Factor-alpha; D064569:alpha7 Nicotinic Acetylcholine Receptor; D009538:Nicotine