Effect of nicotinic acetylcholine receptor α7 subunit gene on liver inflammatory reaction in mice with nonalcoholic steatohepatitis and related mechanisms
10.3760/cma.j.issn.1007-3418.2016.10.010
- VernacularTitle:烟碱型乙酰胆碱受体α7基因对非酒精性脂肪性肝炎小鼠肝脏炎症反应的作用及其机制
- Author:
Fuqiang LI
1
;
Xiaocui WU
;
Lina XU
;
Xiaomei CHEN
;
Si LU
;
Cuilan TANG
Author Information
1. 312030,绍兴市中心医院感染科
- Keywords:
Fatty liver;
α7-AChR gene knockout;
Kupffer cells
- From:
Chinese Journal of Hepatology
2016;24(10):767-771
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of nicotinic acetylcholine receptor αt7 (a7nAChR) 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 supematant;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 1334200,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.
