α7烟碱型乙酰胆碱受体调控小鼠肺泡巨噬细胞中脂多糖诱导炎性反应  

α7 nicotinic acetylcholine receptor mediates lipopolysaccharide -induced inflammatory response in the mouse macrophages

作　　者：付朝晖[1] 余愿[1] 袁世荧[1] 姚尚龙[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,武汉430022

出　　处：《中华实验外科杂志》2015年第12期3082-3085,共4页Chinese Journal of Experimental Surgery

基　　金：湖北省科技厅自然科学基金资助项目（2014CFB444）

摘　　要：目的探讨胡烟碱型乙酰胆碱受体（cJnAChR）调控MH—S细胞中脂多糖（LPS）诱导炎性反应及其作用机制。方法体外培养MH—S细胞：（1）LPS刺激，检测a7nAChR表达；（2）LPS、GTS-21、维库溴铵（Vecuronium）、LPS＋GTS-21、LPS＋Vecuronium、LPS＋GTS-21＋Vecuronium分别刺激，检测肿瘤坏死因子-α（TNF-α）、高迁移率族蛋白B1（HMGBl）、白细胞介素（IL）-6和IL-10释放；（3）下调a7nAChR表达后，重复步骤（2）；（4）LPS、LPS＋GTS-21分别刺激，检测p65核因子-κB（NF-κB）及磷酸化转录信号转导子与激活子3（pSTAT3）表达。结果LPS刺激后，α7nAChRmRNA和蛋白表达分别增加124％和88％（P〈0．05），TNF-α、HMGBl、IL-6和IL-10释放分别增加35．7、18．5、17．4和16．8倍（P〈0．05）；通过GTS-21＋LPS刺激，TNF-α、IL-6和HMGBl释放分别减少64％、56％和61％（P〈0．05），IL-10释放增加13％（P〉0．05）；联用LPS＋GTS-21＋Vecuronium刺激，TNF-α、IL-6和HMGB1的释放分别减少53％、50％和51％（P〈0．05），IL—10的释放增加11％（P〉0．05）；下调α7nAChR后，再通过GTS-214-LPS刺激，TNF-α、IL-6和HMGBl的释放分别减少4．0％、0．3％和12．0％（P〉0．05）；激活a7nAChR后，LPS诱导的p65NFκB表达下降57％（P〈0．05），而pSTAT3活性上升130％（P〈0．05）。结论LPS可以上调α7nAChR表达，释放TNF-α、HMGBI、IL-6和IL-10；激活胡nAchR可以抑制TNF-α、IL-6和HMGBl释放，此抑制作用不能被Vecuronium阻断，但是可以通过下调αTnAChR表达解除；这种抑制作用可能是通过抑制细胞内NF-κB激活和促进酪氨酸激酶酪氨酸激酶／信号转导2（JAK2）-转录信号转导子与激活子3（STAT3）活化实现的。Objective To investigate the inhibitory effects and related mechanisms of stimulation of α7 nicotinic acetylcholine receptor （ctTnAChR） on lipopolysaccharide （ LPS ） - induced inflammatory response in the mouse alveolar macrophages. Methods MH - S cells were cultured. （ 1 ） Cells were trea- ted with LPS, and the expression of α7nAChR was detected; （2） Cells were treated with LPS, GTS -21 , Vecuronium, LPS ＋ GTS - 21, LPS ＋ Vecuronium and LPS ＋ GTS - 21 ＋ Vecuronium separately, and the release of tumor necrosis factor -α （ TNF - α）, high mobility group protein B1 （ HMGB1 ） , interleukin （IL） -6 and IL- 10 was measured; （3） The expression of cOnAChR was knocked down by specific small interfering RNA （siRNA） before the same stimulations and measurements as above; （4） Activity of nucle- ar factor- κB （NF - κB ） and janus kinase （JAK） - signal transducers and activators of transcription 3 （STAT3） was measured after treatment with GTS -21. Results Expression levels of α7nAChR mRNA and protein were increased by about 124% and 88% （P 〈0. 05） after LPS stimulation, and the release of TNF -α, HMGB1, IL - 6 and IL - 10 was increased by 35.7, 18.5, 17.4 and 16. 8 folds separately （P 〈0. 05）. The release of TNF - a, IL -6 and HMGB1 was reduced by about 64% , 56% and 61% respectively（P 〈0. 05 ）, and the release of IL - 10 increased by approximately 13% （ P 〉 0. 05 ） after GTS -21 and LPS co - stimulation. When co - treatment with LPS ＋ GTS -21 ＋ Vecuronium, LPS - in- duced TNF - ct, IL -6 and HMGB1 release was reduced by about 53% , 50% and 51% （P 〈0. 05） , and the release of IL- 10 increased by approximately 11% （P 〉 0. 05 ） , respectively. When the expression ofα7nAChR was knocked down by specific siRNA, TNF - α, IL - 6 and HMGB1 release was reduced by about 4. 0% , 0. 3% and 12. 0% when co - treatment with LPS ＋ GTS - 21 ＋ Vecuronium （ P 〉 0. 05 ）. After co - treatment with LPS and GTS -21, the expression

关 键 词：Α7烟碱型乙酰胆碱受体 胆碱能抗炎反应通路 肺泡巨噬细胞 炎性反应 脂多糖 

分 类 号：R749.16[医药卫生—神经病学与精神病学]